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Effect and underlying mechanism of Huangjing Qianshi decoction in pre-diabetes mouse model

BMC Complementary Medicine and Therapies volume 25, Article number: 151 (2025) Cite this article

Abstract

Background

Insulin secretion deficiency and increased insulin resistance are key pathological pathways that lead to pre-diabetes. Without intervention, pre-diabetes can easily develop into type 2 diabetes mellitus. However, no specific medicine is available for treating pre-diabetes except for intervention through lifestyle changes. Huangjing Qianshi decoction (HJQST) is a qi-replenishing and yin-nourishing Chinese medicinal compound. However, the mode and mechanism of action of HJQST in improving pre-diabetes remain unclear. Here, we studied the effect of HJQST on pre-diabetes.

Methods

BKS-db mice were induced to develop pre-diabetes and treated with HJQST and metformin (MET). After treatment for 51 days, hematoxylin-eosin and oil red O staining were used to analyze the pathological damage and lipid droplet formation in the pancreatic, liver and skeletal muscle of pre-diabetic mice. Serum levels of free fat acid (FFA), glycated hemoglobin A1c (HbA1c), fasting insulin (INS), reactive oxygen species (ROS), and tumor necrosis factor-α (TNF-α) were analyzed. Levels of glucose transporter 4 (GLUT-4), INS, nuclear receptor subfamily 3 group c member 2 (NR3C2), phosphorylated-signal transducer and activator of transcription 1 (p-STAT1), peroxisome proliferator activated receptor co-activator 1 α (PGC-1α), and protein inhibitor of activated STAT1 (PIAS1) protein were analyzed by immunohistochemistry and western blot.

Results

The body weight, fasting blood glucose (FBS) levels, and serum levels of HbA1c, FFA, ROS, and TNF-α were significantly decreased, whereas the insulin level was significantly increased in pre-diabetic BKS-db mice after HJQST treatment. Additionally, HJQST treatment improved pancreatic and liver damage and the lipid droplet formation in liver and skeletal muscle. Furthermore, the increased NR3C2 and p-STAT1 protein levels and decreased GLUT-4, INS, PIAS1, and PGC-1a protein levels in pre-diabetic mice were reversed by HJQST treatment.

Conclusion

HJQST treatment could reverse high FBS level and aberrant lipid metabolism, oxidative stress, and inflammation in pre-diabetes, all of which are related to the NR3C2/PIAS1/STAT1/PGC-1α signal axis.

Peer Review reports

Background

Pre-diabetes, also known as impaired glucose regulation, refers to the state wherein normal glucose regulation develops into impaired glucose regulation, and blood glucose rises without reaching the diagnostic criteria for diabetes (fasting blood glucose [FBS]: 6.1–7 mmol/L or 2-h plasma glucose after oral glucose challenge: 7.8–11.1 mmol/L) [1]. The prevalence of pre-diabetes is on the rise worldwide and is estimated to affect approximately 552 million people by 2030 [2]. Frequently, pre-diabetes can develop into type 2 diabetes mellitus (T2DM). Research data show that the total prevalence of T2DM among adults in China is 8.4%, and 27.6% of them develop from pre-diabetes [3]. Pre-diabetes is generally asymptomatic and difficult to detect. Most patients discover it during physical examinations or due to other diseases. However, pre-diabetes is a reversible disease. Early intervention in the population with pre-diabetes can significantly reduce the probability of them developing T2DM [4, 5]. Therefore, effectively reversing the pre-diabetes status is of great significance in reducing the occurrence of T2DM.

Preventing the progression of pre-diabetes mainly relies on lifestyle changes, including weight loss and exercise, or metformin (MET) treatment, although lifestyle changes have greater benefits than MET treatment [6]. Currently, no targeted drug for pre-diabetes is available. Traditional Chinese medicine has been proven to effectively treat pre-diabetes, diabetes, and their related complications. Huoxue Jiangtang drinks can slow the transition from pre-diabetes to diabetes [7]. Jin-tang-ning improved hyperglycemia and pre-diabetic development by regulating mitochondrial metabolism and endoplasmic reticulum stress of β cells [8]. Traditional Chinese medicine treatment combined with lifestyle changes can significantly regulate the progress of pre-diabetes [9, 10]. Additionally, qi-replenishing Chinese medicine is effective and safe for delaying or reversing pre-diabetes [11]. Huangjing Qianshi decoction (HJQST) is a qi-replenishing and yin-nourishing Chinese medicinal compound. HJQST can effectively improve the FBS levels of patients with pre-diabetes, with significant clinical effects. However, the mode and mechanism of action of HJQST in improving pre-diabetes remain unclear.

Pre-diabetes includes impaired fasting glucose and impaired glucose tolerance, and insulin secretion deficiency and increased insulin resistance (IR) are key pathological pathways that leading to pre-diabetes. The onset of IR is earlier than that of T2DM and even earlier than that of pre-diabetes [12]. The liver and skeletal muscles are key organs that regulate fat and glucose metabolism, and both exhibit IR during the progression of pre-diabetes [13, 14]. Inflammation is also closely associated with pre-diabetes. The serum tumor necrosis factor a (TNF-α) level in population with pre-diabetes including those with impaired fasting glucose and impaired glucose tolerance was significantly increased, whereas the serum interleukin (IL)-6 level showed no significant difference [15]. The pancreatic levels of IL-1β, IL-6, and TNF-α were increased in those with pre-diabetes than in healthy individuals [16]. Oxidative stress is another important factor affecting the development of pre-diabetes [17]. Hyperglycemia can upregulate the markers of chronic inflammation and oxidative stress, which in turn, can lead to IR and impaired insulin secretion. Inhibition of excessive production of inflammation and reactive oxygen species (ROS) is crucial for delaying pre-diabetes progression to diabetes [18].

Signal transducer and activator of transcription 1 (STAT1)/ peroxisome proliferator activated receptor co-activator 1 α (PGC-1α) was closely related to diabetes and its complications. STAT1 activation promoted M1 macrophage polarization to enhance chronic inflammation and IR in obese mice [19]. The expression of STAT1 is negatively correlated with FBS level in obese mice and humans, and STAT1 deficiency can enhance mitochondrial function, tricarboxylic acid cycle, and energy metabolism, thereby improving IR [20]. Sanye tablet reduced IR in obese mice via reducing STAT1 to inhibit JAK/STAT signaling pathway [21]. STAT1 inhibited PGC-1α transcription by binding to the PGC-1α promoter in mitochondria, leading to mitochondrial electron transport chain defect and dysfunction [22]. PGC-1α protein was reduced in pre-diabetes mice, and upregulation PGC-1α alleviated skeletal muscle metabolic disorders [23]. Hence, STAT1/PGC-1α can become a target to improve the pre-diabetes state.

Here, we analyzed the effects of HJQST treatment on FBS and blood lipid levels and tissue damage in pre-diabetic mice. We then studied the effects of HJQST treatment on the expression of glucose transporter 4 (GLUT-4) in the liver and skeletal muscle, fasting insulin (INS) in the pancreatic tissue, and inflammation and oxidative stress in pre-diabetic mice. In addition, transcriptome sequencing revealed that nuclear receptor subfamily 3 group member 2 (NR3C2) mRNA expression was significantly higher in pre-diabetic mice than that in the normal mice, and network pharmacological analysis and transcriptome sequencing revealed that HJQST treatment targeted and inhibited the NR3C2 mRNA expression in our previous study [24, 25], indicating that NR3C2 may be a target for HJQST treatment. Therefore, this study further analyzed the effect of HJQST treatment on the NR3C2 protein and STAT1/PGC-1α signaling pathways in pre-diabetic mice and preliminarily explored the mechanism of HJQST treatment to improve the development of pre-diabetes.

Materials and methods

Animals

Forty specific pathogen-free BKS-db mice (BKS-Leprem2Cd479/Nju gene knockout mice, resulting in spontaneous blood sugar elevation by knocking out the leptin receptor Lepr, Strain NO.T002407, four-week-old, 18–22 g) and six wild-type C57BLKS/JGpt mice (WT group, four-week-old, 10–13 g) were purchased from the GemPharmatech (Nanjing, China) and bred in the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine [SYXK(Xiang)2020-0010]. The animals were adaptively fed with conventional feeding for 1 week, irradiated with a 60 W fluorescent lamp (07:00–21:00), and indoor temperature (20 ± 3 ℃). The experimental protocol was approved by the Ethics Committee of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine (approval number: ZYFY20220615-23) in accordance with ARRIVE guidelines. FBS level in the mice was measured every 3 days. When the FBS level was between 8 and 11 mmol/L for a continuous week (about 6 weeks-old), the animal was considered a pre-diabetic mouse [26]. Then, these mice began to receive treatment.

Treatment

Thirty pre-diabetic mice were randomly divided into five groups: control group (BKS-db + saline), low-/middle-/high-dose HJQST treatment (BKS-db + L-/M-/H- HJQST), and MET treatment (BKS-db + MET). The HJQST dosage for humans is 150 g/day, including dry rhizomes of Polygonatum cyrtonema Hua (Huangjing), 15 g; dry seed of Euryles Semen (Qianshi): 30 g; dry rhizomes of Rhizoma Dioscoreae (Shanyao), 15 g; dry rhizomes of Radix Paeoniae Alba (Baishao), 15 g, dry seed of Jujubae Fructus (Dazao), 7 pieces (about 39 g); Pseudostellariae Radix (Taizisheng), 30 g; dry aboveground parts of Eupatorium fortunei Turcz. Herba Eupatorii (Peilanye), 6 g [25]. It converted to the mouse dosage was 30.8 g/kg (12.33 × 150 g/60 kg). Hence, the mice in the BKS-db + L-/M-/H- HJQST groups were administered 15.4 (30.8/2), 30.8, and 61.6 (30.8 × 2) g/kg HJQST by gavage, respectively. Mice in the BKS-db + MET group were administered 200 mg/kg MET by gavage [27], whereas those in the BKS-db + saline group were administered the same volume of saline by gavage. Finally, the C57BLKS/JGpt mice in the WT group were administered the same volume of saline by gavage. Body weight and FBS levels were recorded every 3 days. After 51 days of treatment, 130 mg/kg pentobarbital was used to euthanize the mice, and those were ensured death by cervical dislocation. Serum and tissue samples were collected from the pancreas, liver, and skeletal muscle. HJQST preparation: Huangjing (lot No. 2012002 C), Qianshi (lot No. 2104001 S), Shanyao (lot No. 2106006 S), Baishao (lot No. 2101012 S), Dazao (lot No. 1912003 S), Taizisheng (lot No. 2103001 S), Peilanye (lot No. 2102002 C) were purchased from CR SANJIU (Guangzhou, China). Three times the amount of water was add to traditional Chinese medicine and soaked for 1 h, then decocted in water for 50 min. The operation was repeated once. Then two liquids were combined and concentrated to 50 ml (Concentration: 3 g/ml, equivalent to containing 3 g of raw medicine per milliliter of liquid medicine). The volume of gastric lavage in the L-/M-/H- HJQST groups is 15.4, 30.8, and 61.6 g/kg × the body weight of mice (kg) / 3 (g/ml), respectively. All analysis personnel are unaware of the animal grouping situation.

Oral glucose tolerance test

All mice were orally administered 1 g/kg body weight of glucose (dissolved in distilled water to 10%w/v) after over-night fasting for oral glucose tolerance test (OGTT) analysis. Blood glucose levels were measured at 0, 30, 60, and 120 min after orally glucose.

Hematoxylin-eosin and oil red O staining

A hematoxylin-eosin (HE) staining kit and oil red O saturated solution (Solarbio, Beijing, China) were used to analyze the pathological damage and lipid droplet formation in pre-diabetic mice. The procedure was performed according to the manufacturer’s instructions.

Enzyme-linked immunosorbent assay (ELISA) and colorimetric assay

Serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and free fat acid (FFA) levels were analyzed using colorimetric assay kits (E-BC-K109-M, E-BC-K261-M, E-BC-K221-M, and E-BC-K013-M, respectively, Elabscience, Wuhan, China). Serum levels of glycated hemoglobin A1c (HbA1c, ab285317, Abcam), INS (E-EL-M2614c, Elabscience), ROS (E-BC-K138-F, Elabscience), and TNF-a (KE10002, Proteintech, Wuhan, China) were analyzed using ELISA. Protein concentration was determined using a microplate reader (VersaMax 384, Molecular Devices, Sunnyvale, CA, USA).

Immunohistochemistry (IHC) and Western blot

Glucose transporter 4 (GLUT-4), INS, NR3C2, and the protein inhibitor of activated STAT1 (PIAS1) levels were analyzed using IHC. In addition, NR3C2, PIAS1, STAT1, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-Alpha (PGC-1a), and GAPDH protein levels were analyzed using western blotting [28]. The PVDF film was place in the ChemiScope 6100 chemiluminescence gel imaging system (CLINX, Shanghai, China), start the exposure, adjust the brightness and contrast, take pictures, and save the western blot image. GAPDH served as the reference protein. Antibodies against GLUT-4 (1:100 dilution, 66846-1-Ig), INS (1:1000 dilution, 66198-1-Ig), NR3C2 (1:2000 dilution for western blot and 1:100 dilution for IHC, 21854-1-AP), PIAS1 (1:2000 dilution for western blot and 1:100 dilution for IHC, 23395-1-AP), STAT1 (1:2000 dilution, 66545-1-Ig), p-STAT1 (1:2000 dilution, 28977-1-AP), PGC-1a (1:5000 dilution, 66369-1-Ig), and GAPDH (1:20000 dilution, 60004-1-Ig) were purchased from the Proteintech (Wuhan Sanying, Wuhan, China).

Statistical analysis

All statistical analyses were performed using the Statistical Package for Social Sciences (SPSS) version 19.0 (IBM Inc.). All data were normally distributed and are presented as means ± standard deviation (SD). Differences between groups were analyzed using one-way analysis of variance (ANOVA), followed by Dunnett’s multiple comparison post-hoc test. Body weight and blood glucose were analyzed using a mixed-ANOVA. *p < 0.05, **p < 0.01, and ***p < 0.001 were used to show statistical significance.

Results

HJQST treatment reduced blood glucose and increased insulin levels in pre-diabetic mice

Compared with those in the control BKS-WT mice, the body weight, FBS, OGTT, and serum HbA1c levels in pre-diabetic BKS-db mice were significantly increased, whereas those in the M-HJQST, H-HJQST, and MET treated-pre-diabetic mice were significantly reduced (Fig. 1A and D). However, the effects of M-HJQST, H-HJQST, and MET treatment on body weight, FBS, OGTT, and HbA1c levels in pre-diabetic mice were not significantly different (Fig. 1A and D). Furthermore, compared with that in the control BKS-WT mice, insulin level was significantly reduced in pre-diabetic BKS-db mice, which can be increased in M-HJQST and H-HJQST group and unaffected in MET group (Fig. 1E).

Fig. 1
figure 1

Huangjing Qianshi decoction (HJQST) treatment reduced body weight (A), fasting blood glucose (FBS) level (B), oral glucose tolerance test (C), and glycated hemoglobin (HbA1c) level (D) while increasing the insulin level (E) in pre-diabetic mice. *p < 0.05, **p < 0.01, and ***p < 0.001

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HJQST treatment reduced FFA levels in pre-diabetic mice

TC, TG, and FFA levels were significantly increased, whereas HDL-C levels were significantly decreased in pre-diabetic BKS-db mice than in the control BKS-WT mice. However, MET treatment significantly reduced TC levels, whereas M-HJQST and H-HJQST treatment significantly reduced FFA levels (Fig. 2). HJQST and MET treatment had no significant effect on TG and HDL-C levels (Fig. 2). Body weight, FBS, and serum HbA1c, INS, TC, TG, HDL-C, and FFA had no significantly changed between M-HJQST and H-HJQST group (Figs. 1 and 2). Hence, M-HJQST treatment group was selected for further pathological and mechanistic studies.

Fig. 2
figure 2

Blood lipids levels were regulated after Huangjing Qianshi decoction (HJQST) and MET treatment. TC: total cholesterol; TG: triglyceride; HDL-C: high-density lipoprotein cholesterol; FFA: free fatty acid. *p < 0.05, **p < 0.01, and ***p < 0.001

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HJQST treatment improved pathological damage and inhibited lipid droplet formation in pre-diabetic mice

HE staining showed that the pancreatic islet cells in BKS-WT mice were arranged in an orderly manner, with a clear morphology and abundant cytoplasm. However, the pancreatic tissue in pre-diabetic BKS-db mice showed morphological changes and decreased cytoplasm in pancreatic islet cells and mild dilation of blood vessels. Both M-HJQST and MET treatments improved pancreatic injury to a certain extent (Fig. 3A). Liver cells were arranged neatly, tightly, and radially in BKS-WT mice, whereas the liver cytoplasm in pre-diabetic BKS-db mice showed vacuolization, increased nuclear pyknosis, disordered arrangement, and infiltration of inflammatory cells. M-HJQST and MET treatment improved liver injury to a certain extent, with M-HJQST having a better effect on improving pathological damage than that of MET treatment (Fig. 3A). However, there were no significant differences in the changes of skeletal muscle tissue among the four groups (Fig. 3A). Furthermore, oil red O staining showed increased lipid droplet formation in pre-diabetic BKS-db mice than in BKS-WT mice, and M-HJQST and MET treatment obviously inhibited lipid droplet formation (Fig. 3B). Compared with that in the BKS-WT mice, GLUT-4 protein level in the pre-diabetes BKS-db mice was obviously reduced, and M-HJQST and MET treatment obviously enhanced GLUT-4 protein level (Fig. 3C). Compared with that in the BKS-WT mice, INS protein level in the pre-diabetes BKS-DB mice was obviously reduced, and M-HJQST and MET treatment obviously enhanced INS protein (Fig. 3D). MET treatment had a greater effect on the increase of GLUT-4 and INS protein levels than M-HJQST treatment (Fig. 3C and D).

Fig. 3
figure 3

Huangjing Qianshi decoction (HJQST) treatment improved the pathological damage and inhibited lipid droplet formation in pre-diabetic mice. (A) hematoxylin and eosin staining were used to detect the pathological morphology of the tissue of pre-diabetic mice after M-HJQST and MET treatment. (B) Lipid accumulation in the liver and skeletal muscle was detected using oil red O staining after M-HJQST and MET treatment. (C) glucose transporter 2/4 (GLUT-2/4) protein level in the liver and skeletal muscle was detected using immunohistochemistry after M-HJQST and MET treatment. (D) Fasting insulin (INS) protein in the pancreas was detected using immunohistochemistry after M-HJQST and MET treatment. Magnification ×100

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HJQST treatment reduced oxidative stress and inflammation in pre-diabetic mice

Compared with those in the BKS-WT group, ROS and TNF-α levels in the BKS-db mice were significantly increased, and M-HJQST and MET treatment reduced the levels of both (Fig. 4).

Fig. 4
figure 4

Reactive oxygen species (ROS) and tumor necrosis factor-alpha (TNF-α) levels in the serum of pre-diabetic mice were reduced after Huangjing Qianshi decoction (HJQST) and MET treatment. *p < 0.05, **p < 0.01, and ***p < 0.001

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HJQST treatment reduced NR3C2 and increased PIAS1 protein levels in pre-diabetic mice

To investigate the mechanism of action of HJQST, the protein levels of NR3C2 and PIAS1 in the pancreatic, liver, and skeletal muscle tissues were analyzed using IHC and western blot.NR3C2 protein level was obviously increased, and PIAS1 protein level was obviously decreased in BKS-db mice compared with in the BKS-WT mice, and M-HJQST and MET treatment obviously reversed the expression trend of both proteins (Figs. 5 and 6).

Fig. 5
figure 5

NR3C2 protein level in the tissues of pre-diabetic mice was reduced, whereas protein inhibitor of activated STAT1 (PIAS1) protein level was increased after Huangjing Qianshi decoction (HJQST) and MET treatment. PIAS1 (A) and NR3C2 (B) proteins were measured using immunohistochemistry. Magnification ×100

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HJQST treatment silenced the STAT1/PGC-1α pathway in pre-diabetic mice

To investigate the effect of HJQST treatment on signaling pathways, the protein levels of STAT1, p-STAT1, and PGC-1α in liver and skeletal muscle tissues were analyzed using western blot. p-STAT1 protein level was significantly increased, and PGC-1α protein was significantly decreased in BKS-db mice compared with in the BKS-WT mice, and M-HJQST and MET treatment significantly reversed the expression trend of both proteins (Fig. 6).

Fig. 6
figure 6

NR3C2, protein inhibitor of activated STAT1 (PIAS1), p-STAT1, and PGC-1α protein level was affected in the tissues of pre-diabetic mice after Huangjing Qianshi decoction (HJQST) and MET treatment. Proteins in the liver tissue and skeletal muscle tissues of pre-diabetic mice after Huangjing Qianshi decoction (HJQST) and MET treatment were measured using western blot. Full-length blots are presented in Supplementary Fig. 1. *p < 0.05, **p < 0.01, and ***p < 0.001

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Discussion

Without intervention the pre-diabetes, it is easy to develop into T2DM. However, pre-diabetes treatment had no specific medication except for intervention through lifestyle change. In this study, we studied the beneficial effect of HJQST in pre-diabetes. In population with pre-diabetes, FBS level is significantly increased, whereas INS level is significantly decreased; both FBS and INS levels could be reversed via intervention [29]. Individuals with high HbA1c level in pre-diabetes are more likely to develop T2DM [30]. The present study showed significantly increased weight, FBS, OGTT, and HbA1c levels and significantly decreased INS levels in pre-diabetic mice. Moreover, HJQST and MET interventions reversed these findings, showing that HJQST intervention could delay the progression of pre-diabetes. In addition, HJQST treatments improved pancreatic and liver tissues injury and increased GLUT-4 expression, indicated that HJQST treatment can improve insulin secretion, glucose transfer, and liver and pancreatic tissue loss, and had a good therapeutic effect.

In addition, Lipid droplet formation was increased in pre-diabetic BKS-db mice and HJQST and MET treatment significantly inhibited lipid droplet formation. Abnormal lipid metabolism, including hypertriglyceridemia and high serum FFA concentration, is one of the main factors leading to IR, pre-diabetes, and T2DM [31]. A high FFA level is a key regulatory factor in IR [32]. FFA and TG levels were significantly reduced in patients with pre-diabetes [31]. The present study showed lipid disturbances including high FFA, TG, and TC levels and low HDL-C levels in pre-diabetic mice, suggesting that the pre-diabetic mice had abnormal lipid metabolism. Although HJQST treatment reduced FFA, TG, and TC levels and increased HDL-C levels, only FFA reduction showed significant differences. The specific reason is still unclear. This may be related to the short treatment time. Subsequent experiments to further study the effects of a combination intervention involving lifestyle and HJQST on blood glucose and lipid levels are warranted.

Population with pre-diabetes is characterized by insulin secretion disorder and IR, which are closely related to TNF-α accumulation [33]. An elevated TNF-α level inhibits GLUT-4 protein and INS-regulated glucose transport, causing the elevation of blood glucose level [34]. In addition, TNF-α disturbs the synthesis and secretion of insulin, inducing the aging and apoptosis of β-cells in pancreatic islets [33]. Freshwater clam extracts delay the progression of pre-diabetes by targeting TNF-α [35]. The presence of ROS is an important marker of oxidative stress, and its level is significantly increased in patients with pre-diabetes. Habit intervention can reduce ROS levels and blood sugar level and reverse pre-diabetes state [36]. Previous studies have shown that ROS and TNF-α levels are significantly increased in pre-diabetic mice. However, HJQST treatment significantly reduced ROS and TNF-α levels, suggesting that HJQST could improve oxidative stress and inflammation in pre-diabetic mice.

This study found that NR3C2 and p-STAT1 protein levels were enhanced, whereas PIAS1 and PGC-1α protein levels were downregulated in the liver, pancreas, and skeletal muscle tissues of pre-diabetic mice. Serum NR3C2 levels are elevated in patients with T2DM [37]. High NR3C2 expression induces oxidative stress and inflammation, which are among the most important causes of IR [38, 39]. NR3C2 is also a potential target of Chuanxiong Rhizoma in treating diabetic nephropathy [40]. A previous study showed that NR3C2 can directly bind to PIAS1 to regulate PIAS1 protein [41]. Overexpression of PIAS1 can improve obesity-induced IR and alleviate diabetic peripheral neuropathy, indicating that PIAS1 is related to inflammation and IR [42, 43]. The PIAS1 protein functions as a SUMO E3 ligase, which is considered an inhibitor of STAT1 by mediating the SUMOylation of STAT1 DNA [44, 45]. In high-fat diet-induced obese mice, high STAT1 expression is closely related to IR and mitochondrial function [20, 21]. STAT1 inhibited PGC-1α transcription to induce mitochondrial electron transport chain defect and dysfunction [22]. The above research and the present study indicated that the NR3C2/PIAS1/STAT1/PGC-1α signal axis is closely related to IR and pre-diabetes. In the present study, NR3C2 and STAT1 protein levels were reduced, whereas PIAS1 and PGC-1α protein levels were increased in pre-diabetic mice after HJQST treatment. Thus, the mechanism of HJQST intervention in reversing the occurrence of pre-diabetes is related to the NR3C2/PIAS1/STAT1/PGC-1 α signal axis.

Overall, the addition of M-HJQST treatment produce a similar treatment effect as MET. MET has good therapeutic effects in terms of blood glucose control. However, it had some side effects: gastrointestinal reactions, such as stomach pain, reluctance to eat, nausea, and poor appetite, diarrhoea, dyspepsia [46, 47]; In addition, a few patients may also experience liver and kidney function damage, erectile dysfunction, and vitamin B12 level reduction [48, 49]. HJQST is a qi-replenishing and yin-nourishing Chinese medicinal compound. We found that two patients with pre-diabetes (5%) occurred side effects after HJQST treatment, both of which were tolerably mild gastrointestinal discomfort in our previous research [50]. Therefore, HJQST can become a substitute for metformin to treat patients with pre-diabetes.

However, there are several limitations to this study. Firstly, the target cells (such as liver cells and skeletal muscle cells) regulated by HJQST are not yet clear. Secondly, this project only clarifies the regulatory role of HJQST on NR3C2 expression, and its regulatory mechanism still needs further clarification. Third, the expression of NR3C2 in patients with pre-diabetes is still unclear, and its regulatory role and mechanism on pre-diabetes need to be further clarified. Therefore, this project will further study the regulatory effect and mechanism of NR3C2 on pre-diabetes, and the role and mechanism of HJQST in reversing pre-diabetes through NR3C2. Finally, abnormal increase in blood sugar can lead to abnormal lipid metabolism, and this study found that MET and HJQST treatment only improved FBS and OGTT, but did not significantly improve blood lipids, especially TG and HDL-C. Previous studies have shown that the absence of leptin receptor mice exhibit hepatic lipid accumulation, even when maintained on a standard diet. This suggests that the leptin receptor is a key regulator of lipid metabolism [51, 52]. Therefore, the absence of leptin receptors might be the reason why HJQST treatment does not show any effect on lipid improvements. However, the reason for this still needs further research. Next, from a genetic perspective, while BKS-db mice are capable of producing leptin, they lack its primary receptor, resulting in a high appetite characteristic of this genetic model of diabetes. High appetite leads to obesity, hyperglycemia, and insulin resistance, which is considered the good obesity complication, pre-diabetes, and T2DM model [53,54,55]. High growth rate of BKS-db occurs between 4 and 10 weeks of age and coincides with an early phase of hyperinsulinemia [56, 57]. Compared to C57BL mice6, BKS-db mice can spontaneously develop obesity, hyperglycemia, and hyperinsulinemia under the normal diet without any additional treatment. Additionally, blood glucose levels in BKS- db mice continued to rise from 4 weeks of age, maintaining hyperglycemia throughout their lifespan. The hyperglycemic phenotype cannot be improved spontaneously, making these mice more suitable for studying the effects of antihyperglycemic drugs. Furthermore, BKS-db mice exhibit greater accumulation of saturated and monounsaturated fatty acids in non-adipose tissues, including the eyes, liver, skeletal muscle, and pancreas [58]. Furthermore, early hepatic IR manifests prior to the onset of diabetes [54]. Therefore, this study selects BKS-db mice to explore the mechanisms underlying insulin resistance, elevated blood glucose levels, and ectopic lipid accumulation in the prediabetic state. However, it remains unclear whether NR3C2 functions exclusively in BKS-db pre-diabetic mice or if it also plays an important role in other pre-diabetic mouse models. According to previous research literature, animals with leptin receptor knockout initially exhibit a state of hyperinsulinemia, followed by a state of hypoinsulinemia later on. Therefore, changes in insulin levels may be a dynamic process [59, 60]. However, it is not clear whether hyperinsulinemia or hypoinsulinemia occurs in pre-diabetes model mice in this study.

Conclusion

HJQST treatment could reverse elevated FBS level, lipid metabolism, oxidative stress, and inflammation in pre-diabetes, which were related to the NR3C2/PIAS1/STAT1/PGC-1 α signal axis. Individuals with pre-diabetes could benefit from HJQST intervention, which might provide a new dietary avenue and drug treatment for delaying diabetes onset.

Data availability

The datasets used or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

IR:

insulin resistance

T2DM:

Type 2 diabetes mellitus

HJQST:

Huangjing Qianshi decoction

MET:

Metformin

FFA:

Free fat acid

HbA1c:

Glycated hemoglobin A1c

INS:

Fasting insulin

ROS:

Reactive oxygen species

TNF-α:

Tumor necrosis factor-α

GLUT-4:

Glucose transporter 4

NR3C2:

Nuclear receptor subfamily 3 group c member 2

p-STAT1:

Phosphorylated-signal transducer and activator of transcription 1

PGC-1α:

Peroxisome proliferator activated receptor co-activator 1 α

PIAS1:

Protein inhibitor of activated STAT1

FBS:

Fasting blood glucose

OGTT:

Oral glucose tolerance test

References

  1. 2. Classification and diagnosis of diabetes: standards of medical care in Diabetes-2021. Diabetes Care. 2021;44(Suppl 1):S15–33.

    Google Scholar 

  2. Teoh KW, Ng CM, Chong CW, Bell S, Cheong WL, Lee SWH. Knowledge, attitude, and practice toward pre-diabetes among the public, patients with pre-diabetes and healthcare professionals: a systematic review. BMJ Open Diabetes Res Care. 2023;11(1):e003203.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Song Y, Zhang X, Zhang H, Yang Q, Zhang S, Zhang Y, Chen Y, Ji Y, Hu X. Prevalence of diabetes and prediabetes in adults from a Third-Tier City in Eastern China: A Cross-Sectional study. Diabetes Therapy: Res Treat Educ Diabetes Relat Disorders. 2019;10(4):1473–85.

    Article  CAS  Google Scholar 

  4. Luo X, Wang Z, Li B, Zhang X, Li X. Effect of resistance vs. aerobic exercise in pre-diabetes: an RCT. Trials. 2023;24(1):110.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Mirmiran P, Hosseini S, Bahadoran Z, Azizi F. Dietary pattern scores in relation to pre-diabetes regression to normal glycemia or progression to type 2 diabetes: a 9-year follow-up. BMC Endocr Disorders. 2023;23(1):20.

    Article  CAS  Google Scholar 

  6. Echouffo-Tcheugui JB, Perreault L, Ji L, Dagogo-Jack S. Diagnosis and management of prediabetes: A review. JAMA. 2023;329(14):1206–16.

    Article  PubMed  Google Scholar 

  7. Zhang PX, Zeng L, Meng L, Li HL, Zhao HX, Liu DL. Observation on clinical effect of Huoxue-Jiangtang Decoction formula granules in treating prediabetes: a randomized prospective placebo-controlled double-blind trial protocol. BMC Complement Med Ther. 2022;22(1):274.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Liu SN, Liu Q, Lei L, Sun SJ, Li CN, Huan Y, Hou SC, Shen ZF. The Chinese patent medicine, Jin-tang-ning, ameliorates hyperglycemia through improving Β cell function in pre-diabetic KKAy mice. Chin J Nat Med. 2020;18(11):827–36.

    CAS  PubMed  Google Scholar 

  9. Jiang L, Wang S, Zhao J, Chien C, Zhang Y, Su G, Chen X, Song D, Chen Y, Huang W, et al. Treatment options of traditional Chinese patent medicines for dyslipidemia in patients with prediabetes: A systematic review and network meta-analysis. Front Pharmacol. 2022;13:942563.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Jiang L, Zhang Y, Zhang H, Chen Y, Huang W, Xiao Y, Aijia Shen E, Li Z, Xue T, Zhao J, et al. Comparative efficacy of 6 traditional Chinese patent medicines combined with lifestyle modification in patients with prediabetes: A network meta-analysis. Diabetes Res Clin Pract. 2022;188:109878.

    Article  PubMed  Google Scholar 

  11. Xia S, Gao B, Chen S, Lin X, Zhang P, Chai Y, Li C, Asakawa T. Verification of the Efficacy and Safety of Qi-Replenishing Chinese Medicine in Treating Prediabetes: A Meta-Analysis and Literature Review. Evidence-based complementary and alternative medicine: eCAM. 2020;2020:7676281.

  12. Khan RMM, Chua ZJY, Tan JC, Yang Y, Liao Z, Zhao Y. From Pre-Diabetes to diabetes: diagnosis, treatments and translational research. Med (Kaunas Lithuania). 2019;55(9):546.

    Google Scholar 

  13. Calderón-DuPont D, Torre-Villalvazo I, Díaz-Villaseñor A. Is insulin resistance tissue-dependent and substrate-specific? The role of white adipose tissue and skeletal muscle. Biochimie. 2023;204:48–68.

    Article  PubMed  Google Scholar 

  14. Balakrishnan R, Thurmond DC. Mechanisms by which skeletal muscle myokines ameliorate insulin resistance. Int J Mol Sci 2022, 23(9).

  15. Onalan E, Bozkurt A, Gursu MF, Yakar B, Donder E. Role of betatrophin and inflammation markers in type 2 diabetes mellitus, prediabetes and metabolic syndrome. J Coll Physicians Surgeons–Pakistan: JCPSP. 2022;32(3):303–7.

    Article  Google Scholar 

  16. Weaver JR, Odanga JJ, Breathwaite EK, Treadwell ML, Murchinson AC, Walters G, Fuentes DP, Lee JB. An increase in inflammation and islet dysfunction is a feature of prediabetes. Diab/Metab Res Rev. 2021;37(6):e3405.

    Article  CAS  Google Scholar 

  17. Dimova R, Chakarova N, Grozeva G, Kirilov G, Tankova T. The relationship between glucose variability and insulin sensitivity and oxidative stress in subjects with prediabetes. Diabetes Res Clin Pract. 2019;158:107911.

    Article  CAS  PubMed  Google Scholar 

  18. Luc K, Schramm-Luc A, Guzik TJ, Mikolajczyk TP. Oxidative stress and inflammatory markers in prediabetes and diabetes. J Physiol Pharmacology: Official J Pol Physiological Soc 2019, 70(6).

  19. Li Y, Dong X, He W, Quan H, Chen K, Cen C, Wei W. Ube2L6 promotes M1 macrophage polarization in High-Fat Diet-Fed obese mice via isgylation of STAT1 to trigger STAT1 activation. Obes Facts. 2024;17(1):24–36.

    Article  CAS  PubMed  Google Scholar 

  20. Singh N, Lawana V, Luo J, Phong P, Abdalla A, Palanisamy B, Rokad D, Sarkar S, Jin H, Anantharam V, et al. Organophosphate pesticide Chlorpyrifos impairs STAT1 signaling to induce dopaminergic neurotoxicity: implications for mitochondria mediated oxidative stress signaling events. Neurobiol Dis. 2018;117:82–113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Yao M, Li L, Huang M, Tan Y, Shang Y, Meng X, Pang Y, Xu H, Zhao X, Lei W, et al. Sanye tablet ameliorates insulin resistance and dysregulated lipid metabolism in High-Fat Diet-Induced obese mice. Front Pharmacol. 2021;12:713750.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Sisler JD, Morgan M, Raje V, Grande RC, Derecka M, Meier J, Cantwell M, Szczepanek K, Korzun WJ, Lesnefsky EJ, et al. The signal transducer and activator of transcription 1 (STAT1) inhibits mitochondrial biogenesis in liver and fatty acid oxidation in adipocytes. PLoS ONE. 2015;10(12):e0144444.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Tian Y, Shi D, Liao H, Lu B, Pang Z. The role of Huidouba in regulating skeletal muscle metabolic disorders in prediabetic mice through AMPK/PGC-1α/PPARα pathway. Diabetol Metab Syndr. 2023;15(1):145.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Cai JL, Li XP, Zhu YL, Deng GM, Yang L, Xia XH, Yi GQ, Chen XY. [Mechanism of Huangjing Qianshi Decoction in treatment of prediabetes based on network Pharmacology and molecular docking]. Zhongguo Zhong Yao Za zhi = Zhongguo Zhongyao Zazhi = China J Chin Materia Med. 2022;47(4):1039–50.

    Google Scholar 

  25. Cai JL, Zhu YL, Li XP, Xia XH, Deng GM, Tong QZ, Yi GQ, Cheng B. [Mechanism of Huangjing Qianshi Decoction in treatment of prediabetic mice based on transcriptome sequencing]. Zhongguo Zhong Yao Za zhi = Zhongguo Zhongyao Zazhi = China J Chin Materia Med. 2023;48(4):1032–42.

    Google Scholar 

  26. Lee KL, Aitken JF, Li X, Montgomery K, Hsu HL, Williams GM, Brimble MA, Cooper GJS. Vesiculin derived from IGF-II drives increased islet cell mass in a mouse model of pre-diabetes. Islets. 2022;14(1):14–22.

    Article  CAS  PubMed  Google Scholar 

  27. Gali S, Kundu A, Sharma S, Ahn MY, Puia Z, Kumar V, Kim IS, Kwak JH, Palit P, Kim HS. Therapeutic potential of bark extracts from Macaranga denticulata on renal fibrosis in streptozotocin-induced diabetic rats. J Toxicol Environ Health Part A. 2024;87(23):911–33.

    Article  CAS  Google Scholar 

  28. Cai JL, Li XP, Zhu YL, Yi GQ, Wang W, Chen XY, Deng GM, Yang L, Cai HZ, Tong QZ, et al. Polygonatum sibiricum polysaccharides (PSP) improve the palmitic acid (PA)-induced Inhibition of survival, inflammation, and glucose uptake in skeletal muscle cells. Bioengineered. 2021;12(2):10147–59.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. AkbariRad M, Shariatmaghani SS, Razavi BM, Majd HM, Shakhsemampour Z, Sarabi M, Jafari M, Azarkar S, Ghalibaf AM, Khorasani ZM. Probiotics for glycemic and lipid profile control of the pre-diabetic patients: a randomized, double-blinded, placebo-controlled clinical trial study. Diabetol Metab Syndr. 2023;15(1):71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Halalau A, Roy S, Hegde A, Khanal S, Langnas E, Raja M, Homayouni R. Risk factors associated with glycated hemoglobin A1c trajectories progressing to type 2 diabetes. Ann Med. 2023;55(1):371–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Ha X, Cai X, Cao H, Li J, Yang B, Jiang R, Li X, Li B, Xin Y. Docking protein 1 and free fatty acids are associated with insulin resistance in patients with type 2 diabetes mellitus. J Int Med Res. 2021;49(11):3000605211048293.

    Article  CAS  PubMed  Google Scholar 

  32. A ISS, C AB. A JS: Changes in plasma free fatty acids associated with Type-2 diabetes. Nutrients. 2019;11(9).

  33. Liu Y, Ma C, Li P, Ma C, He S, Ping F, Zhang H, Li W, Xu L, Li Y. Potential Protective Effect of Dietary Intake of Non-α-Tocopherols on Cellular Aging Markers Mediated by Tumor Necrosis Factor-α in Prediabetes: A Cross-Sectional Study of Chinese Adults. Oxidative medicine and cellular longevity 2020, 2020:7396801.

  34. Akash MSH, Rehman K, Liaqat A. Tumor necrosis Factor-Alpha: role in development of insulin resistance and pathogenesis of type 2 diabetes mellitus. J Cell Biochem. 2018;119(1):105–10.

    Article  CAS  PubMed  Google Scholar 

  35. Huang TH, Ke CH, Chen CC, Chuang CH, Liao KW, Shiao YH, Lin CS. The effects of freshwater clam (Corbicula fluminea) extract on serum tumor necrosis Factor-Alpha (TNF-α) in prediabetic patients in Taiwan. Mar Drugs 2022, 20(4).

  36. La Sala L, Tagliabue E, Mrakic-Sposta S, Uccellatore AC, Senesi P, Terruzzi I, Trabucchi E, Rossi-Bernardi L, Luzi L. Lower miR-21/ROS/HNE levels associate with lower glycemia after habit-intervention: DIAPASON study 1-year later. Cardiovasc Diabetol. 2022;21(1):35.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Cao Z, Yao F, Lang Y, Feng X. Elevated Circulating LINC-P21 serves as a diagnostic biomarker of type 2 diabetes mellitus and regulates pancreatic β-cell function by sponging miR-766-3p to upregulate NR3C2. Experimental and clinical endocrinology & diabetes: official journal. German Soc Endocrinol [and] German Diabetes Association. 2022;130(3):156–64.

    Article  CAS  Google Scholar 

  38. Huang Y, Wang Y, Ouyang Y. Elevated microRNA-135b-5p relieves neuronal injury and inflammation in post-stroke cognitive impairment by targeting NR3C2. Int J Neurosci. 2022;132(1):58–66.

    Article  CAS  PubMed  Google Scholar 

  39. Wang C, Hu F. Long noncoding RNA SOX2OT Silencing alleviates cerebral ischemia-reperfusion injury via miR-135a-5p-mediated NR3C2 Inhibition. Brain Res Bull. 2021;173:193–202.

    Article  CAS  PubMed  Google Scholar 

  40. Hu S, Chen S, Li Z, Wang Y, Wang Y. Research on the potential mechanism of Chuanxiong rhizoma on treating diabetic nephropathy based on network Pharmacology. Int J Med Sci. 2020;17(15):2240–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. He K, Zhang J, Liu J, Cui Y, Liu LG, Ye S, Ban Q, Pan R, Liu D. Functional genomics study of protein inhibitor of activated STAT1 in mouse hippocampal neuronal cells revealed by RNA sequencing. Aging. 2021;13(6):9011–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Liu Y, Ge X, Dou X, Guo L, Liu Y, Zhou SR, Wei XB, Qian SW, Huang HY, Xu CJ, et al. Protein inhibitor of activated STAT 1 (PIAS1) protects against Obesity-Induced insulin resistance by inhibiting inflammation cascade in adipose tissue. Diabetes. 2015;64(12):4061–74.

    Article  CAS  PubMed  Google Scholar 

  43. Hou Z, Chen J, Yang H, Hu X, Yang F. PIAS1 alleviates diabetic peripheral neuropathy through sumolation of PPAR-γ and miR-124-induced downregulation of EZH2/STAT3. Cell Death Discovery. 2021;7(1):372.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Lu S, Peng X, Zeng S, Deng H, Feng Z, Zeng Q, Cheng X, Hu J, Ye Z, Li M, et al. Grass carp (Ctenopharyngodon idellus) PIAS1 inhibits innate immune response via interacting with STAT1. Dev Comp Immunol. 2021;125:104216.

    Article  PubMed  Google Scholar 

  45. Su X, Zhang Q, Yue J, Wang Y, Zhang Y, Yang R. TRIM59 suppresses NO production by promoting the binding of PIAS1 and STAT1 in macrophages. Int Immunopharmacol. 2020;89(Pt A):107030.

    Article  CAS  PubMed  Google Scholar 

  46. Tarry-Adkins JL, Grant ID, Ozanne SE, Reynolds RM, Aiken CE. Efficacy and side effect profile of different formulations of metformin: A systematic review and Meta-Analysis. Diabetes Therapy: Res Treat Educ Diabetes Relat Disorders. 2021;12(7):1901–14.

    Article  CAS  Google Scholar 

  47. Hameed M, Khan K, Salman S, Mehmood N. Dose comparison and side effect profile of Metformin extended release versus Metformin immediate release. J Ayub Med Coll Abbottabad: JAMC. 2017;29(2):225–9.

    PubMed  Google Scholar 

  48. Ngu MH, Zakaria R, Mohd Zulkifli M, Ab Rahman R. Erectile dysfunction as a possible important side effect of metformin: A case report. Malaysian Family Physician: Official J Acad Family Physicians Malaysia. 2023;18:20.

    Article  Google Scholar 

  49. Yu YM, So SKC, Khallouq BB. The effect of Metformin on vitamin B12 level in pediatric patients. Annals Pediatr Endocrinol Metabolism. 2022;27(3):223–8.

    Article  Google Scholar 

  50. Cai J, Zhu Y, Zhou L, Pan W, Yang H, Yang L, Zhang Z, Tong Q, Li X, Yi G. Clinical effect of Huangjing Qianshi Decoction on prediabetes. Asia-Pacific Traditional Med (Chinese). 2021;17(03):100–3.

    Google Scholar 

  51. Nason SR, Kim T, Antipenko JP, Finan B, DiMarchi R, Hunter CS, Habegger KM. Glucagon-Receptor signaling reverses hepatic steatosis independent of leptin receptor expression. Endocrinology 2020, 161(1).

  52. Huynh FK, Neumann UH, Wang Y, Rodrigues B, Kieffer TJ, Covey SD. A role for hepatic leptin signaling in lipid metabolism via altered very low density lipoprotein composition and liver lipase activity in mice. Hepatology (Baltimore MD). 2013;57(2):543–54.

    Article  CAS  PubMed  Google Scholar 

  53. Zahid S, Dafre AL, Currais A, Yu J, Schubert D, Maher P. The geroprotective drug candidate CMS121 alleviates diabetes, liver inflammation, and renal damage in Db/db leptin receptor deficient mice. Int J Mol Sci. 2023;24(7):6828.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Davis RC, Castellani LW, Hosseini M, Ben-Zeev O, Mao HZ, Weinstein MM, Jung DY, Jun JY, Kim JK, Lusis AJ, et al. Early hepatic insulin resistance precedes the onset of diabetes in obese C57BLKS-db/db mice. Diabetes. 2010;59(7):1616–25.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Wu G, Gu W, Cheng H, Guo H, Li D, Xie Z. Huangshan Maofeng Green Tea Extracts Prevent Obesity-Associated Metabolic Disorders by Maintaining Homeostasis of Gut Microbiota and Hepatic Lipid Classes in Leptin Receptor Knockout Rats. Foods (Basel, Switzerland). 2022;11(19).

  56. Campbell-Tofte J, Mu H, Winther K, Mølgaard P, Belin N, Josefsen K. Standardization parameters and synergism of source plant materials for the antidiabetic efficacy of the Rauvolfia-Citrus tea. Fitoterapia. 2024;176:106004.

    Article  CAS  PubMed  Google Scholar 

  57. Lee GH, Proenca R, Montez JM, Carroll KM, Darvishzadeh JG, Lee JI, Friedman JM. Abnormal splicing of the leptin receptor in diabetic mice. Nature. 1996;379(6566):632–5.

    Article  CAS  PubMed  Google Scholar 

  58. Campbell-Tofte J, Hansen HS, Mu H, Mølgaard P. Increased lipids in non-lipogenic tissues are indicators of the severity of type 2 diabetes in mice. Prostaglandins Leukot Essent Fat Acids. 2007;76(1):9–18.

    Article  CAS  Google Scholar 

  59. Bao D, Ma Y, Zhang X, Guan F, Chen W, Gao K, Qin C, Zhang L. Preliminary characterization of a leptin receptor knockout rat created by CRISPR/Cas9 system. Sci Rep. 2015;5:15942.

    Article  PubMed  PubMed Central  Google Scholar 

  60. Chisada S, Sugiyama A. Renal lesions in leptin receptor-deficient Medaka (Oryzias latipes). J Toxicologic Pathol. 2019;32(4):297–303.

    Article  CAS  Google Scholar 

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Acknowledgements

None.

Funding

This work was supported by the Project of guiding innovation of clinical medical technology in Hunan province, China (2021SK51408), Project of scientific research program of Hunan Health Committee, China (B202303066210), Natural Science Foundation of Hunan Province, China (2023JJ60486 and 2021JJ80068), and Scientific research project of traditional Chinese medicine in Hunan province, China (D2022091).

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Author notes

  1. Jialuo Cai and Yilin Zhu contributed equally to this work.

Authors and Affiliations

  1. Preventive Treatment of Disease Center, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China

    Jialuo Cai & Xiaoping Li

  2. Graduate School of Hunan University of Traditional Chinese Medicine, Changsha, 410208, China

    Yilin Zhu

  3. Department of Scientific Research, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410208, China

    Guiming Deng

  4. Department of Pharmacy, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China

    Linqi Ouyang & Wangzhong Xiao

  5. Department of Health Management, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China

    Fang Zhou

  6. Department of Scientific Research, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China

    Yuanshan Han

  7. Library’s Office, Hunan University of Chinese Medicine, Changsha, 410208, China

    Feiyun Yuan

  8. Preparation Center, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, Hunan, China

    Li Huang

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Contributions

Jialuo Cai, Yilin Zhu, and Xiaoping Li contributed to the conception and design of the work, acquisition of data, analysis and interpretation of data, and drafting of the manuscript. Guiming Deng, Linqi Ouyang, Wangzhong Xiao, Fang Zhou, Yuanshan Han, Feiyun Yuan, and Li Huang contributed to animal experiment and collected the data. All authors have approved this manuscript.

Corresponding author

Correspondence to Xiaoping Li.

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The experimental protocol was approved by the Ethics Committee of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine (approval number: ZYFY20220615-23) in accordance with ARRIVE guidelines. This study did not involve clinical trials; hence, the Clinical Trial Number was not provided in the manuscript. We only mentioned our previous clinical research (reference 50) on HJQST in the manuscript.

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Cai, J., Zhu, Y., Deng, G. et al. Effect and underlying mechanism of Huangjing Qianshi decoction in pre-diabetes mouse model. BMC Complement Med Ther 25, 151 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12906-025-04893-z

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BMC Complementary Medicine and Therapies

ISSN: 2662-7671

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