1.Gelian Tiaotang Pills Ameliorate Renal Fibrosis in db/db Mice via NLRP3/Caspase-1/GSDMD Pathway
Lihui FAN ; Zhigang WANG ; Xia YANG ; Xiaolong MEI ; Kun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):136-145
ObjectiveTo investigate the effect of Gelian Tiaotang pills on renal fibrosis in db/db mice based on the nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3)/cysteinyl aspartate-specific proteinase (Caspase)-1/gasdermin D (GSDMD) signaling pathway. MethodsForty db/db mice were randomly assigned into model, positive control (0.001 3 g·kg·d-1 dapagliflozin suspension), and high-, medium-, and low-dose Gelian Tiaotang pills (3.12, 1.56, and 0.78 g·kg·d-1 suspension of Gelian Tiaotang pills, respectively) groups, with 8 mice in each group. Eight db/m mice were selected as the normal group. The normal group and model group were given equal volumes of pure water, while the drug interventions groups were administrated with corresponding agents by gavage once a day for 12 consecutive weeks. The general conditions of mice were observed daily. The fasting blood glucose (FBG) and body mass were measured every 4 weeks. Kidneys were weighed after sampling, and the kidney index was calculated. An automatic biochemical analyzer was used to measure the serum levels of triglyceride (TG), total cholesterol (TC), serum creatinine (SCr), and blood urea nitrogen (BUN). The pathological changes, extracellular matrix deposition, and renal fibrosis degree were examined by hematoxylin-eosin, periodic acid-schiff (PAS), and Masson staining, respectively. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin (IL)-1β and IL-18 in the renal tissue. Immunohistochemistry (IHC) was used to detect the localization and expression of fibronectin and collagen Ⅰ in the renal tissue. Western blot was employed to determine the protein levels of NLRP3, Caspase-1, cleaved Caspase-1, GSDMD, and GSDMD-N in the renal tissue. ResultsCompared with the normal group, the model group generally had poor general states and increases in the body mass, kidney weight, kidney index, and levels of FBG, TG, TC, SCr, and BUN (P<0.01). In addition, glomerular pyknosis, increased matrix, vacuolar degeneration of renal tubular epithelial cells, and interstitial infiltration of inflammatory cells were observed in the model group (P<0.01), together with rises in the levels of IL-1β and IL-18 in the renal tissue (P<0.01) and up-regulated protein levels of NLRP3, Caspase-1, cleaved Caspase-1, GSDMD, GSDMD-N, fibronectin, and collagen Ⅰ in the renal tissue (P<0.01). Compared with the model group, 12 weeks of drug interventions reduced the body mass, kidney weight, and kidney index and lowered the levels of FBG, TG, TC, SCr, and BUN in the serum and IL-1β and IL-18 in the renal tissue (P<0.05, P<0.01). Furthermore, drug interventions ameliorated the renal lesions and down-regulated the protein levels of NLRP3, Caspase-1, cleaved Caspase-1, GSDMD, GSDMD-N, fibronectin, and collagen Ⅰ in the renal tissue (P<0.05, P<0.01). The high-dose group of Gelian Tiaotang pills had the best effects. ConclusionGelian Tiaotang pills may inhibit pyroptosis and reduce inflammatory responses by regulating the NLRP3/Caspase-1/GSDMD signaling pathway, thus delaying the process of renal fibrosis in diabetes.
2.Mechanism and Combination Therapy of Berberine in Treatment of Nonalcoholic Fatty liver Disease:A Review
Xiaojie WANG ; Heng ZHANG ; Sutong LIU ; Lihui ZHANG ; Wenxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):269-281
Nonalcoholic fatty liver disease(NAFLD) is the most common chronic liver disease in the world. Because of its complex pathogenesis, high clinical prevalence and large population, it poses a great threat and challenge to public health in the world. Therefore, active intervention measures are needed. Currently, western medicine is effective in reducing weight, reducing liver fat content, improving glucose-lipid metabolism and insulin resistance. However, for patients with NAFLD-related fibrosis and cirrhosis, there is still a lack of sufficient histological evidence to support its benefits, and randomized controlled trials are still needed to clarify. Lifestyle intervention is an important cornerstone for the treatment of NAFLD, but there are many problems such as poor implementation and low compliance of patients, and the clinical efficacy is not ideal. Traditional Chinese medicine(TCM) has the significant advantages of multiple pathways and multiple targets. Berberine, the active ingredient of TCM, can interfere with the production of NAFLD from multiple pathways, including increasing energy consumption, weight loss, improving glucose-lipid metabolism, improving insulin resistance, anti-inflammatory, anti-oxidation, regulating intestinal flora, restoring bile acid homeostasis, anti-fibrosis and so on, which can play a positive role in the treatment of NAFLD. At the same time, it was found that the combination of BBR with Chinese and western medicines had significant advantages in promoting drug absorption, improving oral bioavailability, increasing the highest biological distribution in the liver, enhancing the overall therapeutic effect of NAFLD, and reducing adverse drug reactions, which could provide reference for clinical medication.
3.Trends in mortality and life loss of gastric cancer in Wenzhou City from 2014 to 2023
YE Zhenmiao ; FAN Lihui ; JIANG Xuexia ; ZHENG Yuhang ; ZHANG Mohan ; LUO Yongyuan ; XIE Yimin ; LI Huijun ; JIN Xi
Journal of Preventive Medicine 2025;37(3):267-271
Objective:
To investigate the trends in mortality and life loss of gastric cancer in Wenzhou City, Zhejiang Province from 2014 to 2023, so as to provide the evidence for formulating the prevention and control strategy for gastric cancer.
Methods:
The surveillance on causes of death data of permanent residents in Wenzhou City were collected through the Wenzhou Chronic Disease Monitoring and Management Information System from 2014 to 2023. The crude mortality of gastric cancer was calculated, and standardized by the data from the Sixth Chinese National Population Census in 2010. The life loss were measured using potential years of life lost (PYLL) and rate of potential years of life lost (PYLLR). The characteristics of mortality and life loss of gastric cancer in different genders and age groups were described. The trends in mortality and PYLLR of gastric cancer were analyzed using the average annual percent change (AAPC).
Results:
Totally 17 080 deaths were reported due to gastric cancer in Wenzhou City from 2014 to 2023, accounting for 12.58% and ranking third in the order of malignant tumor deaths. The crude mortality of gastric cancer was 20.73/105, and the standardized mortality was 15.22/105, showing decreasing trends (AAPC=-3.311%, -6.470%, both P<0.05). The crude mortality of gastric cancer was 29.22/105 in men and 11.61/105 in women, with standardized mortality rates of 20.81/105 and 8.74/105 (both P<0.05). The crude mortality of gastric cancer appeared a tendency towards a rise with increasing age (P<0.05), reaching the highest rate of 225.88/105 in the group aged 80 to <85 years. The PYLL and PYLLR of gastric cancer were 107 607.50 person-years and 1.37‰. The PYLLR appeared a tendency towards a decline from 2014 to 2023, with AAPC of -6.667% (P<0.05).
Conclusions
The mortality and PYLLR of gastric cancer in Wenzhou City appeared a tendency towards a decline from 2014 to 2023. Men and the elderly populations were the key groups for the prevention and treatment of gastric cancer.
4.Study on the protective efect and mechanism of paeoniflorin on palmitic acid-induced HepG2 cells
Tong LIU ; Shanzheng LI ; Cheng ZHOU ; Sutong LIU ; Lihui ZHANG ; Wenxia ZHAO
Journal of Clinical Hepatology 2025;41(3):499-505
ObjectiveTo investigate the role and mechanism of action of paeoniflorin (PF) in protecting HepG2 cells induced by palmitic acid (PA). MethodsHepG2 cells were stimulated with PA at a concentration of 250 μmol/L to establish a NAFLD model. Compound C at a concentration of 10 μmol/L was used as an inhibitor, and PF at a concentration of 25 μmol/L was used for intervention. The experiment was divided into normal group (CON group) treated with complete culture medium, model group (MOD group) treated with PA, PF treatment group (MOD+PF group) treated with PA and PF, model+inhibitor group (MOD+COM group) treated with PA and Compound C, and model+inhibitor+PF group (MOD+COM+PF group) treated with PA, Compound C, and PF. Kits were used to measure lipid deposition indicators, liver function parameters, oxidative stress indicators, and inflammation indicators; oil red O staining was used to observe lipid deposition; Western Blot was used to measure the protein expression levels of AMPK, SIRT1, PGC-1α, mTOR, Beclin-1, LC3, and P62 in cells. The one-way analysis of variance was used for comparison of quantitative data between groups, while the Tukey’s test was used for comparison between two groups. ResultsCompared with the MOD group, PF improved the levels of TC and TG (P<0.05), reduced the levels of ALT, AST, CRP, TNF-α, IL-1β, and IL-6 (P<0.05), increased the activity of SOD and CAT and the level of GSH, and reduced the level of MDA in cells (all P<0.05). Oil red O staining showed that PF alleviated lipid deposition in cells. Western blot results showed that compared with the MOD group, PF increased the protein expression levels of p-AMPK, SIRT1, PGC-1α, LC3Ⅱ/LC3Ⅰ, and Beclin-1 and reduced the protein expression levels of p-mTOR and P62 (all P<0.05). ConclusionPF can inhibit PA-induced oxidative stress and inflammatory response in HepG2 cells, improve lipid deposition, and promote autophagy via the AMPK/SIRT1/PGC-1α/mTOR signaling pathway.
5.Evaluation of the efficacy and safety of cryopreserved deglycerolized red blood cells infusion based on propensity score matching method
Wei YANG ; Fanfan FU ; Lei NIU ; Tingchen XU ; Xin ZHANG ; Hongmei SHI ; Lihui FU ; Chunya MA ; Yang YU
Chinese Journal of Blood Transfusion 2025;38(4):531-536
[Objective] To compare the efficacy and safety of deglycerolized red blood cells (DRBC) and suspended red blood cells (SRBC) based on the propensity score matching (PSM) method, so as to provide evidence for the rational use of DRBC resources in clinical practice. [Methods] A total of 89 patients who received DRBC transfusion and 2 916 patients who received SRBC transfusion in our hospital from January 2023 to September 2024 were included. A 1∶1 nearest neighbor PSM was used to balance covariates such as gender, age, and body mass index (BMI). The changes of hemoglobin (Hb), red blood cell (RBC) count, hematocrit (HCT), and inflammatory markers such as white blood cell (WBC) count, neutrophil (NE) count, C-reactive protein (CRP), and Interleukin-6(IL-6) in the last 72 hours after transfusion were analyzed by SPSS 26.0 and R software to evaluate clinical efficacy and transfusion safety. [Results] The baseline of the two groups was balanced after PSM (P>0.05). There was no significant difference in the total effective rate between the DRBC group (80.9%) and the SRBC group (86.5%) (P>0.05). In the SRBC group, WBC (×10
/L) increased from 9.634±6.742 to 10.147±6.835, CRP (mg/dL) increased from 5.468±4.647 to 6.174±6.114, and IL-6(pg/mL) decreased from 213.733±587.191 to 157.255±552.626. In the DRBC group, WBC (×10
/L) decreased from 11.123±7.880 to 11.011±8.549, CRP (mg/dL) decreased from 5.729±4.761 to 5.326±4.466, and IL-6(pg/mL) decreased from 238.806±639.060 to 152.255±266.558. Compared with the before treatment, the differences between the SRBC group and DRBC group were not statistically significant (P>0.05). Among all patients included in the statistics, the overall incidence of transfusion adverse reactions was 0.205% (6/2 916) in the SRBC group, and no adverse reactions occurred in the DRBC group. The incidence in the SRBC group was higher than that in the DRBC group. [Conclusion] Based on PSM analysis, there was no significant difference in the efficacy and safety of DRBC transfusion compared with SRBC transfusion, which can provide evidence-based support for routine application.
6.Clinical Effect of Xiaozhi Hugan Capsules in Treatment of Patients with Non-alcoholic Steatohepatitis and Its Impact on Serum IL-6 and MCP-1
Xiaoyan LIU ; Suping MA ; Qingliang MA ; Chunxiao LI ; Lihui ZHANG ; Huaxin CHEN ; Wenxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):185-192
ObjectiveTo observe the clinical effect of Xiaozhi Hugan capsules in treating patients with non-alcoholic steatohepatitis (NASH) combined with phlegm-dampness and blood stasis syndrome and its effects on serum interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1). MethodsA total of 124 patients with NASH combined with phlegm-dampness and blood stasis syndrome who were admitted to the Department of Spleen, Stomach, and Hepatobiliary Diseases, the First Affiliated Hospital of Henan University of Chinese Medicine from July 2020 to December 2022 were selected. According to the random number table method, patients were randomly divided into an observation group (62 cases) and a control group (62 cases). The treatment duration was 6 months. The observation group received Xiaozhi Hugan capsules orally, while the control group received polyene phosphatidylcholine capsules. The efficacy indicators included alanine aminotransferase (ALT), aspartate aminotransferase (AST), controlled attenuation parameter (CAP), liver stiffness measurement (LSM), traditional Chinese medicine (TCM) syndrome scores (discomfort/dull pain/distending pain in liver region, fatigue, etc.), body mass index (BMI), waist-to-height ratio (WHtR), high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC), triglycerides (TG), homeostatic model assessment for insulin resistance (HOMA-IR) [including fasting blood glucose (FBG) and fasting insulin level (INS)], free fatty acids (FFA), IL-6, and MCP-1. Adverse drug reactions were recorded. ResultsAfter treatment, the total effective rate in the observation group was 92.3% (48/52), while that in the control group was 75.5% (39/49). The total effective rate in the observation group was higher than that in the control group (χ2=5.339, P<0.05). After treatment, the TCM syndrome scores in both groups were significantly reduced (P<0.05), and the post-treatment scores in the observation group were better than those in the control group (P<0.05). After treatment, the levels of ALT, AST, TC, FFA, fasting insulin (FINS), HOMA-IR, MCP-1, IL-6, CAP, LSM, BMI, and WHtR were decreased (P<0.05) significantly in both groups, and the observation group showed superior improvement in the above indicators compared to the control group (P<0.05). The observation group exhibited significant reductions in TG and FBG (P<0.05) and an increase in HDL-C (P<0.05), while no significant changes were observed in the control group. The observation group was superior to the control group after treatment (P<0.05). No severe adverse reactions occurred in either group during the treatment. ConclusionXiaozhi Hugan capsules have significant clinical efficacy in treating patients with NASH combined with phlegm-dampness and blood stasis syndrome. It reduces hepatic steatosis, lowers liver stiffness, inhibits the expression of serum inflammatory factors, and alleviates liver inflammation. No obvious adverse reactions occur, suggesting it is suitable for clinical application.
7.Clinical Effect of Xiaozhi Hugan Capsules in Treatment of Patients with Non-alcoholic Steatohepatitis and Its Impact on Serum IL-6 and MCP-1
Xiaoyan LIU ; Suping MA ; Qingliang MA ; Chunxiao LI ; Lihui ZHANG ; Huaxin CHEN ; Wenxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):185-192
ObjectiveTo observe the clinical effect of Xiaozhi Hugan capsules in treating patients with non-alcoholic steatohepatitis (NASH) combined with phlegm-dampness and blood stasis syndrome and its effects on serum interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1). MethodsA total of 124 patients with NASH combined with phlegm-dampness and blood stasis syndrome who were admitted to the Department of Spleen, Stomach, and Hepatobiliary Diseases, the First Affiliated Hospital of Henan University of Chinese Medicine from July 2020 to December 2022 were selected. According to the random number table method, patients were randomly divided into an observation group (62 cases) and a control group (62 cases). The treatment duration was 6 months. The observation group received Xiaozhi Hugan capsules orally, while the control group received polyene phosphatidylcholine capsules. The efficacy indicators included alanine aminotransferase (ALT), aspartate aminotransferase (AST), controlled attenuation parameter (CAP), liver stiffness measurement (LSM), traditional Chinese medicine (TCM) syndrome scores (discomfort/dull pain/distending pain in liver region, fatigue, etc.), body mass index (BMI), waist-to-height ratio (WHtR), high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC), triglycerides (TG), homeostatic model assessment for insulin resistance (HOMA-IR) [including fasting blood glucose (FBG) and fasting insulin level (INS)], free fatty acids (FFA), IL-6, and MCP-1. Adverse drug reactions were recorded. ResultsAfter treatment, the total effective rate in the observation group was 92.3% (48/52), while that in the control group was 75.5% (39/49). The total effective rate in the observation group was higher than that in the control group (χ2=5.339, P<0.05). After treatment, the TCM syndrome scores in both groups were significantly reduced (P<0.05), and the post-treatment scores in the observation group were better than those in the control group (P<0.05). After treatment, the levels of ALT, AST, TC, FFA, fasting insulin (FINS), HOMA-IR, MCP-1, IL-6, CAP, LSM, BMI, and WHtR were decreased (P<0.05) significantly in both groups, and the observation group showed superior improvement in the above indicators compared to the control group (P<0.05). The observation group exhibited significant reductions in TG and FBG (P<0.05) and an increase in HDL-C (P<0.05), while no significant changes were observed in the control group. The observation group was superior to the control group after treatment (P<0.05). No severe adverse reactions occurred in either group during the treatment. ConclusionXiaozhi Hugan capsules have significant clinical efficacy in treating patients with NASH combined with phlegm-dampness and blood stasis syndrome. It reduces hepatic steatosis, lowers liver stiffness, inhibits the expression of serum inflammatory factors, and alleviates liver inflammation. No obvious adverse reactions occur, suggesting it is suitable for clinical application.
8.Association between serum creatinine/cystatin C ratio and nonalcoholic fatty liver disease in adults
Qizhen ZHANG ; Sutong LIU ; Lihui ZHANG ; Yajie GUAN ; Junjiao XU ; Wenxia ZHAO ; Minghao LIU
Journal of Clinical Hepatology 2025;41(6):1083-1089
ObjectiveTo investigate the association between serum creatinine/cystatin C ratio (CCR) and nonalcoholic fatty liver disease (NAFLD) based on the NHANES database, and to evaluate the potential significance of CCR as an indicator reflecting the metabolic status of the body. MethodsBased on the data from the NHANES database in 1999 — 2004, a total of 4 217 participants were enrolled and divided into NAFLD group with 1 726 participants and non-NAFLD group with 2 491 participants. CCR was compared between the two groups, and the association between CCR and NAFLD was analyzed. The Wilcoxon rank-sum test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The multivariate logistic regression model was used to investigate the association between CCR and NAFLD; CCR was divided into 4 groups based on quartiles, and odds ratio (OR) and 95% confidence interval (CI) in the regression model was calculated with the first quartile as reference. In addition, the restricted cubic spline analysis was used to investigate whether there was a non-linear relationship between CCR and NAFLD, and interaction items were introduced into the Logistic regression model to perform an interaction analysis. Subgroup analyses were performed based on the stratification of variables to investigate the difference in the association between CCR and NAFLD in different populations. ResultsThe non-NAFLD group had a significantly higher CCR than the NAFLD group (Z=-4.76,P<0.01). The Logistic regression analysis showed that in model 1 without adjustment of variables, CCR was negatively associated with NAFLD (OR=0.993,95%CI:0.989 — 0.996,P<0.01), and in model 3 with adjustment of all variables, CCR was still negatively associated with NAFLD (OR=0.986,95%CI:0.981 — 0.991,P<0.01). The analysis of CCR based on quartiles showed a significant association between the increase in CCR and the reduction in the risk of NAFLD. In model 3, compared with the individuals with the lowest quartile of CCR, the individuals with the highest quartile of CCR had a significantly lower risk of NAFLD (OR=0.426,95%CI:0.316 — 0.574,P<0.01). Further interaction and subgroup analyses showed that the interaction between CCR and age/sex had a statistical significance (Pinteraction<0.01 and Pinteraction=0.04). The subgroup analysis based on age showed a more significant association between CCR and NAFLD in the middle-aged population (≤60 years) (OR=0.982,95%CI:0.976 — 0.987), and the subgroup analysis based on sex showed a stronger association between CCR and NAFLD in women (OR=0.979,95%CI:0.972 — 0.986). ConclusionThis study shows a significant negative association between CCR and NAFLD, and such association is more significant in middle-aged individuals and women.
9.Clinical Study on Treatment of Non-alcoholic Steatohepatitis Patients with Dyslipidemia by Dizhuo Huayu Prescription with Catgut Embedding Therapy
Xiaoyan LIU ; Dongfang SHANG ; Lihui ZHANG ; Chenlu ZHAO ; Siying WANG ; Huaxin CHEN ; Wenxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):152-159
ObjectiveTo observe the clinical efficacy and safety of Dizhuo Huayu prescription combined with catgut embedding therapy in patients with nonalcoholic steatohepatitis (NASH) and dyslipidemia and explore the effect of the combined therapy on inflammatory cytokines interleukin (IL)-18 and IL-1β. MethodsA total of 82 patients with NASH and dyslipidemia from the Gastroenterology Department of the First Affiliated Hospital of Henan University of Chinese Medicine were randomly divided into a control group and a treatment group, with 41 patients in each group. The control group received Polyene Polyenylphosphatidylcholine Capsules, while the treatment group received Dizhuo Huayu prescription granules combined with catgut embedding. The treatment duration was 24 weeks for both groups. At weeks 0, 12, and 24, the traditional Chinese medicine (TCM) syndrome score, body mass index (BMI), liver fat content assessed by Fibroscan (CAP value), the level of alanine transaminase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and free fatty acid (FFA), and the expression of inflammatory cytokines IL-18 and IL-1β in serum were observed. Adverse reactions in both groups were recorded. ResultsA comparison of the comprehensive therapeutic effects between the two groups after 24 weeks of treatment revealed that the total effective rate was 62.16% (23/37) in the control group and 85.71% (30/35) in the treatment group, with a statistically significant difference (χ2 = 5.14, P<0.05). At weeks 12 and 24 after treatment, the TCM syndrome score, BMI, CAP value, TC, TG, LDL-C, and FFA were all significantly lower in both groups compared to pre-treatment levels, while the HDL-C level significantly increased (P<0.05). The effect was better at week 24 (P<0.05) than at week 12 (P<0.05), and the treatment group showed better outcomes than the control group at weeks 12 and 24 after treatment (P<0.05). After 24 weeks of treatment, both groups exhibited significant reductions in IL-18 and IL-1β levels (P<0.05). The treatment group demonstrated superior efficacy compared to the control group after treatment (P<0.05). Both groups experienced decreases in ALT, AST, and GGT levels after treatment (P<0.05). However, there were no statistically significant differences between the 12-week and 24-week post-treatment values within each group, nor were there significant differences between the two groups. No significant adverse reactions were observed in both groups. ConclusionThe Dizhuo Huayu prescription combined with catgut embedding therapy is safe and effective in treating patients with NASH and dyslipidemia, exhibiting hepatoprotective, anti-inflammatory, lipid-regulating, and weight-reducing effects.
10.Baihe Wuyaotang Ameliorates NAFLD by Enhancing mTOR-mediated Liver Autophagy
Rui WANG ; Tiantian BAN ; Lihui XUE ; Xinyi FENG ; Jiyuan GUO ; Jiaqi LI ; Shenghe JIANG ; Xiaolei HAN ; Baofeng HU ; Wenli ZHANG ; Naijun WU ; Shuang LI ; Yajuan QI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):66-77
ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang (BWT) on non-alcoholic fatty liver disease (NAFLD) and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups: normal control, model, positive drug (pioglitazone hydrochloride 1.95×10-3 g·kg-1), and low-, medium-, and high-dose BWT (1.3,2.5 and 5.1 g·kg-1). Following a 12-week high-fat diet (HFD) inducement, the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week, glucose tolerance (GTT) and insulin tolerance (ITT) tests were conducted. Subsequently, the mice were euthanized for the collection of liver tissue and serum, and the subcutaneous adipose tissue (iWAT) and epididymal adipose tissue (eWAT) were weighed. Serum levels of total triglycerides (TG) and liver function indicators,such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were determined. Histological examinations, including oil red O staining, hematoxylin-eosin (HE) staining, Masson staining, and transmission electron microscopy, were performed to evaluate hepatic lipid deposition, pathological morphology, and ultrastructural changes, respectively. Meanwhile, Western blot and real-time quantitative polymerase chain reaction (Real-time PCR) were employed to analyze alterations, at both gene and protein levels, the insulin signaling pathway molecules, including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 (IRS1/2/Akt/FoxO1), glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase (Pepck) and glucose-6-phosphatase (G6Pase), lipid metabolism-related genes stearoyl-coA desaturase-1 (SCD-1) and carnitine palmitoyltransferase-1 (CPT-1), fibrosis-associated molecules α-smooth muscle actin (α-SMA), type Ⅰ collagen (CollagenⅠ), and the fibrosis canonical signaling pathway transforming growth factor-β1/drosophila mothers against decapentaplegic protein2/3(TGF-β1/p-Smad/Smad2/3), inflammatory factors such as interleukin(IL)-6, IL-8, IL-11, and IL-1β, autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1, and the expression of mammalian target of rapamycin (mTOR). ResultCompared with the model group, BWT reduced the body weight and liver weight of NAFLD mice(P<0.05, P<0.01), inhibited liver lipid accumulation, and reduced the weight of white fat: it reduced the weight of eWAT and iWAT(P<0.05, P<0.01) as well as the serum TG content(P<0.05, P<0.01). BWT improved the liver function as reflected by the reduced ALT and AST content(P<0.05, P<0.01). It improved liver insulin resistance by upregulating IRS2, p-Akt/Akt, p-FoxO1/FoxO1 expressions(P<0.05). Besides, it improved glucose and lipid metabolism disorders: it reduced fasting blood glucose and postprandial blood glucose(P<0.05, P<0.01), improved GTT and ITT(P<0.05, P<0.01), reduced the expression of Pepck, G6Pase, and SCD-1(P<0.01), and increased the expression of CPT-1(P<0.01). The expressions of α-SMA, Collagen1, and TGF-β1 proteins were down-regulated(P<0.05, P<0.01), while the expression of p-Smad/Smad2/3 was downregulated(P<0.05), suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6, IL-8, IL-11 and IL-1β(P<0.01) and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression(P<0.05)while downregulating the expression of p62/SQSTM1 and mTOR(P<0.05). ConclusionBWT ameliorates NAFLD by multifaceted improvements, including improving IR and glucose and lipid metabolism, anti-inflammation, anti-fibrosis, and enhancing autophagy. In particular, BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.


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