1.Clinical Efficacy of Tangshen Dihuang Decoction in Treating Diabetic Kidney Disease with Liver-kidney Yin Deficiency and Blood Stasis Syndrome and Its Impact on Gut Microbiota
Chaomao YANG ; Shunxiao ZHANG ; Zhixin YANG ; Jiandong GAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):171-178
ObjectiveTo observe the clinical efficacy of Tangshen Dihuang decoction in treating diabetic kidney disease (DKD) with liver-kidney Yin deficiency and blood stasis syndrome and its impact on gut microbiota. MethodsA randomized controlled clinical trial was conducted, in which 102 DKD patients with liver-kidney Yin deficiency and blood stasis syndrome were randomly assigned to the Tangshen Dihuang decoction group and the control group. Each group consisted of 51 cases, and the treatment period was 3 months. The primary efficacy indicators included urinary albumin-to-creatinine ratio (UACR), fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 hPBG), glycated hemoglobin (HbA1C), serum creatinine (SCr), blood urea nitrogen (BUN), angiotensinⅡ (AngⅡ), serum cystatin C (Cys-C), urinary N-acetyl-β-D-glucosaminidase (NAG), urinary β2-microglobulin (Uβ2-MB), traditional Chinese medicine (TCM) symptom scores, and gut microbiota. ResultsAfter treatment, the total response rate in the Tangshen Dihuang decoction was 87.23% (41/47), which was higher than that (69.57%, 32/46) in the control group (Z=4.30, P<0.05). After treatment, the TCM symptom scores decreased in both groups (P<0.01) and were lower in the Tangshen Dihuang decoction group than in the control group (P<0.01). After treatment, the control group showed decreases in UACR, Uβ2-MG, AngⅡ, and FBG (P<0.05) as well as 2 hPBG and HbA1C (P<0.01), and no significant differences in BUN, Cys-C, eGFR, SCr, and NAG. The Tangshen Dihuang decoction group showed increased eGFR (P<0.05), declined levels of UACR, BUN, Cys-C, Uβ2-MB, AngⅡ, FBG, 2 hPBG, NAG, and HbA1C (P<0.01), and no significant difference in SCr. The Tangshen Dihuang decoction group had lower BUN (P<0.05), Cys-C (P<0.05), AngⅡ (P<0.05), 2 hPBG (P<0.05), Uβ2-MG (P<0.01), and NAG (P<0.01) and higher eGFR level (P<0.05) than the control group. After treatment, the control group showed declines in Shannon, Observed_species, and Chao1 indices (P<0.05). The samples from both groups showed statistically significant differences in the principal coordinates analysis (PCoA) plot based on Anosim analysis (P<0.05). After treatment, the Tangshen Dihuang decoction group showed decreased relative abundance of Actinobacteria (P<0.05). Moreover, the relative abundance of Actinobacteria was significantly lower in the Tangshen Dihuang decoction group than in the control group (P<0.05). At the genus level, the control group showed decreased relative abundance of Bifidobacterium (P<0.05), and the Tangshen Dihuang decoction group presented increased relative abundance of Bifidobacterium and Blautia_A (P<0.05). After treatment, the Tangshen Dihuang decoction group had higher relative abundance of Bifidobacterium than the control group (P<0.01). ConclusionTangshen Dihuang decoction has a significant therapeutic effect on DKD patients with liver-kidney Yin deficiency and blood stasis syndrome. It can markedly relieve clinical symptoms and reduce proteinuria and postprandial blood glucose by antagonizing the local renin-angiotensin system (RAS) and alleviating gut microbiota dysbiosis.
2.Effect and mechanism of transplantation of human umbilical cord mesenchymal stem cells with overexpression of the Numb gene in treatment of cholestatic liver fibrosis
Shihao ZHANG ; Changqing ZHAO ; Mingyan YANG ; Feifei XING ; Wei LIU ; Gaofeng CHEN ; Jiamei CHEN ; Ping LIU ; Yongping MU
Journal of Clinical Hepatology 2026;42(1):80-89
ObjectiveTo investigate the effect and mechanism of transplantation of human umbilical cord mesenchymal stem cell (hUC-MSC) with overexpression of the Numb gene in the treatment of cholestatic liver fibrosis (CLF). MethodsThe technique of lentiviral transfection was used to induce the overexpression of the Numb gene in hUC-MSC (hUC-MSCNumb-OE), and hUC-MSC transfected with empty vector (hUC-MSCOE-EV) was used as negative control. Bile duct ligation (BDL) was performed to establish a rat model of CLF, and then the rats were randomly divided into BDL group, hUC-MSC group, hUC-MSCOE-EV group, and hUC-MSCNumb-OE group, while a sham-operation group was also established. The rats in the intervention groups were given a single splenic injection of the corresponding cells after BDL, and samples were collected at the end of week 4. Related indicators were measured, including serum biochemistry, liver histopathology, the content of hydroxyproline (Hyp) in the liver, hepatic stellate cell activation, ductular reaction, liver regeneration, and the expression levels of key molecules in the Numb-p53 signaling axis. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the BDL group, the hUC-MSC group and the hUC-MSCOE-EV group had significant reductions in the levels of serum biochemical parameters (aspartate aminotransferase, gamma-glutamyl transpeptidase, total bile acid, total bilirubin, and direct bilirubin), liver fibrosis markers (the content of Hyp and the expression levels of alpha-smooth muscle actin, tumor necrosis factor-α, and transforming growth factor-beta 1), and ductular reaction markers (the expression levels of CK7 and CK19) (all P <0.05), and compared with the hUC-MSCOE-EV group, the hUC-MSCNumb-OE group had significantly greater improvements in the above indicators (all P <0.05). In addition, compared with the hUC-MSCOE-EV group, the hUC-MSCNumb-OE group had significant improvements in the expression levels of liver regeneration-related markers (albumin and hepatocyte nuclear factor 4α) and the molecules associated with the Numb-p53 signaling axis (Numb, pNumb, Mdm2, and p53) (all P <0.05). ConclusionOverexpression of the Numb gene can enhance the therapeutic effect of hUC-MSC on CLF, possibly by activating the Numb-PTBL-p53-HNF4α axis, promoting the hepatic differentiation of hUC-MSCs and subsequently enhancing liver regeneration.
3.Mechanism of Wumeiwan on Inhibiting Fatty Acid Metabolism Reprogramming in Prevention and Treatment of Colorectal Cancer Based on Multi-omics Analysis
Gang XIAO ; Shusen YANG ; Mingming SI ; Yanyan YANG ; Hailiang WEI ; Shuguang YAN ; Hui LUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):21-30
ObjectiveTo investigate the mechanism by which Wumeiwan suppresses the development and progression of colorectal cancer(CRC) through the regulation of fatty acid metabolic reprogramming, thereby providing new experimental evidence for the prevention and treatment of CRC. MethodsA total of 120 C57BL/6 mice were randomly divided into the blank group, model group, Wumeiwan high-, medium-, and low-dose groups(54, 27, 13.5 g·kg-1), and the mesalazine group(0.01 g·kg-1), with 20 mice in each group. Except for the blank group, all mice were subjected to azoxymethane(AOM)/dextran sulfate sodium(DSS) treatment to establish an inflammation-associated CRC model. One week after AOM injection, mice in the treatment groups received intragastric administration of the designated drugs, while the blank and model groups received an equal volume of purified water, continuing until 20 d after the intervention endpoint. Hematoxylin-eosin(HE) staining was used to observe colonic histopathological alterations, and immunohistochemistry for vascular endothelial growth factor(VEGF) was performed to evaluate neovascularization and tumor invasion. Metabolomics combined with Kyoto Encyclopedia of Genes and Genomes(KEGG) and metabolite set enrichment analysis(MSEA) was applied to identify key CRC-related metabolic pathways, which were further validated by transcriptomic Gene Ontology(GO) enrichment and gene heatmap analysis. Subsequently, Western blot was performed to determine the expression levels of core proteins in these pathways, and immunofluorescence was used to analyze their localization and co-expression patterns in tissues, thereby elucidating the mechanism of Wumeiwan from multiple biological dimensions. ResultsCompared with the blank group, mice in the model group exhibited a significant decrease in body weight and a significant increase in the disease activity index(DAI) score(P<0.05), with pronounced colonic mucosal damage accompanied by aggravated tumor invasion. Compared with the model group, Wumeiwan intervention markedly improved body weight loss and reduced DAI score, attenuated mucosal injury, and significantly decreased VEGF expression level(P<0.05). Multi-omics analysis revealed that differential metabolites and genes across groups were commonly enriched in fatty acid metabolism, fatty acid biosynthesis, and other lipid-related pathways. Relative to the blank group, the model group showed significant upregulation levels of fatty acid synthesis-related genes, including sterol regulatory element-binding protein 1(SREBP1), fatty acid synthase(FASN), stearoyl-CoA desaturase 1(SCD1), as well as saturated fatty acids(P<0.05). Compared with the model group, treatment with Wumeiwan significantly reduced the expression of key genes involved in fatty acid metabolic pathways, including SREBP1, FASN, and SCD1(P<0.05). Western blot results further confirmed that proteins in this pathway were significantly elevated in the model group, whereas they were markedly downregulated following Wumeiwan treatment(P<0.05). Immunofluorescence analysis demonstrated enhanced co-localization of SREBP1 with the cancer-associated fibroblast(CAF) marker α-smooth muscle actin(SMA) in the model group, whereas this co-localization signal was attenuated after Wumeiwan intervention(P<0.05). ConclusionWumeiwan can improve survival outcomes and alleviate colonic pathological damage in CRC mice, its therapeutic mechanism may be closely associated with the regulation of fatty acid metabolic reprogramming mediated by the SREBP1/FASN/SCD1 signaling pathway.
4.Current analysis of bloodstream infections in adult intensive care unit patients: a multi-center cohort study of China.
Shuguang YANG ; Yao SUN ; Ting WANG ; Hua ZHANG ; Wei SUN ; Youzhong AN ; Huiying ZHAO
Chinese Critical Care Medicine 2025;37(3):232-236
OBJECTIVE:
To analyze the clinical characteristics, microbiological analysis, and drug resistance patterns of intensive care unit (ICU) bloodstream infection.
METHODS:
A prospective cohort study method was employed to collect clinical data from patients suspected of bloodstream infection (BSI) during their stay in ICUs across 67 hospitals in 16 provinces and cities nationwide, from July 1, 2021, to December 31, 2022. Electronic data collection technology was used to gather general information on ICU patients, including gender, age, length of hospital stay, as well as diagnostic results, laboratory tests, imaging studies, microbiological results (including smear, culture results, and pathogen high-throughput testing), and prognosis. Patients were divided into a BSI group and a non-BSI group based on the presence or absence of BSI; further, patients with BSI were categorized into a drug-resistant group and a non-drug-resistant group based on the presence or absence of drug resistance. Differences in the aforementioned indicators between groups were analyzed and compared; variables with P < 0.10 in the univariate analysis were included in a multivariate Logistic regression analysis to identify risk factors for mortality and drug resistance in ICU patients with BSI.
RESULTS:
A total of 2 962 ICU patients suspected of BSI participated in the study, including 790 in the BSI group and 2 172 in the non-BSI group. Patients in the BSI group were mainly from East China and Southwest China, with significantly higher age and mortality rates than those in the non-BSI group. Among ICU patients with BSI, Staphylococcus had the highest detection rate (8.10%), followed by Klebsiella pneumoniae (7.47%); there were 169 cases in the drug-resistant group and 621 cases in the non-drug-resistant group; 666 cases survived, and 124 cases died (mortality was 15.70%). There were statistically significant differences between the death group and the survival group in terms of age, regional distribution, and bloodstream infections caused by Gram negative (G-) bacilli, Enterococcus faecium, Aspergillus, and Klebsiella pneumoniae; multivariate Logistic regression analysis showed that age [odds ratio (OR) = 1.01, 95% confidence interval (95%CI) was 1.00-1.03], regional distribution (OR = 4.07, 95%CI was 1.02-1.34), Enterococcus faecium infection (OR = 3.64, 95%CI was 1.16-11.45), and Klebsiella pneumoniae infection (OR = 2.64,95%CI was 1.45-4.80) were independent risk factors for death in ICU patients with BSI (all P < 0.05). There were statistically significant differences between the drug-resistant group and the non-drug-resistant group in terms of age and bloodstream infections caused by Gram positive (G+) cocci and G- bacilli; multivariate Logistic regression analysis showed that age (OR = 1.01,95%CI was 1.00-1.03), G- bacilli infection (OR = 2.18, 95%CI was 1.33-3.59), Escherichia coli infection (OR = 0.28,95%CI was 0.09-0.84), and Enterococcus faecium infection (OR = 3.35, 95%CI was 1.06-10.58) were independent risk factors for drug resistance in ICU patients with BSI (all P < 0.05).
CONCLUSIONS
Bloodstream infections may increase the mortality of ICU patients. Older age, regional distribution, Enterococcus faecium infection and Klebsiella pneumoniae infection can increase the mortality rate of ICU patients with BSI; bloodstream infections caused by G- bacilli are prone to drug resistance, but have no significant impact on the mortality of ICU patients with BSI.
Adult
;
Humans
;
Bacteremia/microbiology*
;
China/epidemiology*
;
Cohort Studies
;
Cross Infection/microbiology*
;
Drug Resistance, Bacterial
;
Intensive Care Units/statistics & numerical data*
;
Prospective Studies
;
Risk Factors
;
Sepsis/microbiology*
5.Three-dimensional kinematic analysis can improve the efficacy of acupoint selection for post-stroke patients with upper limb spastic paresis: A randomized controlled trial.
Xin-Yun HUANG ; Ou-Ping LIAO ; Shu-Yun JIANG ; Ji-Ming TAO ; Yang LI ; Xiao-Ying LU ; Yi-Ying LI ; Ci WANG ; Jing LI ; Xiao-Peng MA
Journal of Integrative Medicine 2025;23(1):15-24
BACKGROUND:
China is seeing a growing demand for rehabilitation treatments for post-stroke upper limb spastic paresis (PSSP-UL). Although acupuncture is known to be effective for PSSP-UL, there is room to enhance its efficacy.
OBJECTIVE:
This study explored a semi-personalized acupuncture approach for PSSP-UL that used three-dimensional kinematic analysis (3DKA) results to select additional acupoints, and investigated the feasibility, efficacy and safety of this approach.
DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS:
This single-blind, single-center, randomized, controlled trial involved 74 participants who experienced a first-ever ischemic or hemorrhagic stroke with spastic upper limb paresis. The participants were then randomly assigned to the intervention group or the control group in a 1:1 ratio. Both groups received conventional treatments and acupuncture treatment 5 days a week for 4 weeks. The main acupoints in both groups were the same, while participants in the intervention group received additional acupoints selected on the basis of 3DKA results. Follow-up assessments were conducted for 8 weeks after the treatment.
MAIN OUTCOME MEASURES:
The primary outcome was the Fugl-Meyer Assessment for Upper Extremity (FMA-UE) response rate (≥ 6-point change) at week 4. Secondary outcomes included changes in motor function (FMA-UE), Brunnstrom recovery stage (BRS), manual muscle test (MMT), spasticity (Modified Ashworth Scale, MAS), and activities of daily life (Modified Barthel Index, MBI) at week 4 and week 12.
RESULTS:
Sixty-four participants completed the trial and underwent analyses. Compared with control group, the intervention group exhibited a significantly higher FMA-UE response rate at week 4 (χ2 = 5.479, P = 0.019) and greater improvements in FMA-UE at both week 4 and week 12 (both P < 0.001). The intervention group also showed bigger improvements from baseline in the MMT grades for shoulder adduction and elbow flexion at weeks 4 and 12 as well as thumb adduction at week 4 (P = 0.007, P = 0.049, P = 0.019, P = 0.008, P = 0.029, respectively). The intervention group showed a better change in the MBI at both week 4 and week 12 (P = 0.004 and P = 0.010, respectively). Although the intervention group had a higher BRS for the hand at week 12 (P = 0.041), no intergroup differences were observed at week 4 (all P > 0.05). The two groups showed no differences in MAS grades as well as in BRS for the arm at weeks 4 and 12 (all P > 0.05).
CONCLUSION:
Semi-personalized acupuncture prescription based on 3DKA results significantly improved motor function, muscle strength, and activities of daily living in patients with PSSP-UL.
TRIAL REGISTRATION
Chinese Clinical Trial Registry ChiCTR2200056216. Please cite this article as: Huang XY, Liao OP, Jiang SY, Tao JM, Li Y, Lu XY, Li YY, Wang C, Li J, Ma XP. Three-dimensional kinematic analysis can improve the efficacy of acupoint selection for post-stroke patients with upper limb spastic paresis: A randomized controlled trial. J Integr Med. 2025; 23(1): 15-24.
Humans
;
Male
;
Female
;
Middle Aged
;
Acupuncture Points
;
Upper Extremity/physiopathology*
;
Biomechanical Phenomena
;
Single-Blind Method
;
Aged
;
Stroke/therapy*
;
Acupuncture Therapy/methods*
;
Stroke Rehabilitation/methods*
;
Adult
;
Muscle Spasticity/therapy*
;
Paresis/physiopathology*
;
Treatment Outcome
6.Quality control protocol for adult overweight and obesity screening in health management (examination) institutions (2025 edition)
Jianling FAN ; Tiejun WANG ; Pengfei YANG ; Keke DING ; Xiaoning HAO ; Sunfang JIANG ; Ankang LÜ ; Jianping LU ; Sheng RONG ; Weibin SHI ; Shengwei SUN ; Yan TAN ; Qilei TU ; Zhiping WANG ; Bing WANG ; Jianyun WANG ; Weijian WANG ; Yan WANG ; Qun XU ; Chenli ZHANG ; Fan ZHANG ; Ping ZHANG ; Yansong ZHENG ; Jieru ZHOU ; Dan CHEN ; Jiaoyang ZHENG
Chinese Journal of Clinical Medicine 2025;32(6):1097-1111
Obesity, as a chronic recurrent disease, has become a major public health challenge in China. To implement the requirements of the Healthy China Initiative (2019—2030), under domestic guidelines or consensus statements on overweight and obesity, and in alignment with the latest scientific advances globally, the Quality control protocol for adult overweight and obesity screening in health management (examination) institutions (2025 edition) was developed. This protocol was drafted by the Health Management Center of Shanghai Changzheng Hospital and formulated through multiple rounds of deliberation by experts in China’s health examination quality control field. The protocol establishes unified standards for screening facilities, personnel qualifications, and measurement or testing procedures. It defines specific screening items, outlines a standardized screening pathway, and sets requirements for the final medical review, ensuring the scientific validity, effectiveness, and safety of the screening process. The implementation of this protocol will enhance the consistency of weight management practices for adults across health examination institutions and strengthen the quality control of overweight and obesity screening programs.
7.Mahuang Lianqiao Chixiaodou Decoction and its active components inhibit alternative pathway complement activation in rat model of IgA nephropathy.
Ting SONG ; Guang-Yu SHENG ; Wei RUAN ; Ya-Heng ZHANG ; Xue-Jun YANG
China Journal of Chinese Materia Medica 2025;50(6):1626-1636
This study aims to investigate the mechanism of Mahuang Lianqiao Chixiaodou Decoction(MHLQ) and its main active components in treating immunoglobin A nephropathy(IgAN). The rat model of IgAN was established by a combination of measures including gavage of bovine serum albumin, subcutaneous injection of carbon tetrachloride, and tail vein injection of lipopolysaccharide. The modeled rats were randomized into model, low-, medium-, and high-dose(1.773, 3.545, and 7.090 g·kg~(-1), respectively) MHLQ, phillyrin(PHI, 0.020 g·kg~(-1)), pseudoephedrine(PSE, 0.020 g·kg~(-1)), and losartan potassium(LP, 9.003 mg·kg~(-1)) groups, and Wistar rats were used as the control. Rats were administrated with corresponding drugs by gavage, and those in the control and model groups received an equal volume of normal saline. All the groups were treated for 4 consecutive weeks. Urine, serum, liver, and kidney samples were collected from rats in each group at the end of drug administration. The 24 h urine protein and renal function were examined, and staining was performed to observe the pathological changes in the renal tissue. The immunofluorescence assay was employed to detect the expression of IgA and complement C3/C3b/C3c in the renal tissue. Electron microscopy was employed to observe the ultrastructure of the renal tissue. Enzyme-linked immunosorbent assay was performed to determine the expression of complement C3 and sublytic C5b-9 in the serum and renal tissue. Western blot was performed to determine the expression levels of hepatic and renal complement C3/C3b/C3c, C5/C5a, C5b-9, and complement factor B(CFB). Immunohistochemistry(IHC) was employed to measure the expression of complement C3 in the renal tissue. The results showed that compared with the control group, the model group had elevated levels of blood urea nitrogen and serum creatinine, proliferation of glomerular mesangial cells and extracellular matrix, and glomerular deposition of IgA immune complexes or electron-dense material. In addition, the model group showcased increased serum C3 levels and up-regulated expression of CFB, C3/C3b/C3c, C5/C5a, and C5b-9 in the renal tissue and C3/C3b/C3c and C5b-9 in the hepatic tissue. After treatment with MHLQ and its active components, all of the above indexes were reversed. In conclusion, MHLQ and its active components can improve the renal function and reduce the deposition of immune complexes and pathological damage in the renal tissue of the rat model of IgAN by inhibiting the alternative pathway complement activation.
Animals
;
Drugs, Chinese Herbal/administration & dosage*
;
Glomerulonephritis, IGA/genetics*
;
Rats
;
Male
;
Disease Models, Animal
;
Rats, Wistar
;
Complement Activation/drug effects*
;
Kidney/immunology*
;
Humans
8.Expert consensus on evaluation index system construction for new traditional Chinese medicine(TCM) from TCM clinical practice in medical institutions.
Li LIU ; Lei ZHANG ; Wei-An YUAN ; Zhong-Qi YANG ; Jun-Hua ZHANG ; Bao-He WANG ; Si-Yuan HU ; Zu-Guang YE ; Ling HAN ; Yue-Hua ZHOU ; Zi-Feng YANG ; Rui GAO ; Ming YANG ; Ting WANG ; Jie-Lai XIA ; Shi-Shan YU ; Xiao-Hui FAN ; Hua HUA ; Jia HE ; Yin LU ; Zhong WANG ; Jin-Hui DOU ; Geng LI ; Yu DONG ; Hao YU ; Li-Ping QU ; Jian-Yuan TANG
China Journal of Chinese Materia Medica 2025;50(12):3474-3482
Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards*
;
Humans
;
Consensus
;
Drugs, Chinese Herbal/therapeutic use*
;
Surveys and Questionnaires
9.Changes in hepatic bile acid profile in a mouse model of metabolic-associated steatohepatitis induced by a high-fat, high-sugar, and high-cholesterol diet combined with carbon tetrachloride
Jingjing WANG ; Jinghua PENG ; Yu LIU ; Feipeng XU ; Wei LIU ; Hailin YANG ; Ping LIU
Journal of Clinical Hepatology 2025;41(4):661-669
ObjectiveTo compare the hepatic bile acid profile between a mouse model of metabolic-associated steatohepatitis (MASH) induced by a high-fat, high-sugar, and high-cholesterol diet combined with intraperitoneal injection of 10% carbon tetrachloride (CCl4) and MASH cases in clinical practice, and to investigate the feasibility of this model in studying drug interventions on bile acid profile in MASH. MethodsA total of 30 male C57BL/6J mice were randomly divided into control group and model group, with 15 mice in each group. The mice in the control group were given normal diet and drinking water and weekly injections of olive oil, and those in the model group were given a high-fat, high-sugar, and high-cholesterol diet, high-sugar drinking water, and weekly injections of CCl4+olive oil. At the end of weeks 8, 12, and 16, 5 mice were selected from each group to collect samples. Behavioral assessments were performed, and body weight and liver wet weight were measured; liver pathology and lipid deposition were evaluated by HE staining, SAF scoring, oil Red O staining, the semi-quantitative analysis of stained area, the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and liver triglyceride (TG) content; Sirius Red staining was performed for liver tissue to assess liver fibrosis; ultra-performance liquid chromatography-tandem mass spectrometry and targeted metabolomics were used to measure the hepatic bile acid profile, including cholic acid (CA), glycocholic acid (GCA), chenodeoxycholic acid (CDCA), glycochenodeoxycholic acid (GCDCA), ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (TUDCA), hyodeoxycholic acid (HDCA), and glycodeoxycholic acid (GDCA). The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsCompared with the control group at the same time point, the model group had disheveled and dull fur, reduced activity, and relatively slow reactions at weeks 8, 12, and 16, as well as significant increases in liver wet weight (P<0.05), the serum level of ALT (P<0.05), the content of TG in the liver (P<0.05), and SAF score (P<0.05). As for the differentially expressed bile acids in liver tissue, compared with the control group at week 8, the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA, TUDCA, HDCA, and GDCA (all P<0.05); compared with the control group at week 12, the model group had significantly higher levels of CA, GCA, CDCA, and GCDCA and significantly lower levels of UDCA and HDCA (all P<0.05); compared with the control group at week 16, the model group had significantly higher levels of CA, GCA, CDCA, GCDCA, and TUDCA and significantly lower levels of UDCA, HDCA, and GDCA (all P<0.05). As for the differentially expressed bile acids in the bile acid pool of liver tissue, compared with the control group at week 8, the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA, TUDCA, GDCA, and HDCA (all P<0.05); compared with the control group at weeks 12 and 16, the model group had significantly higher levels of GCA and GCDCA and significantly lower levels of UDCA, GDCA, and HDCA (all P<0.05). ConclusionThere are significant changes in the hepatic bile acid profile in a mouse model of MASH induced by a high-fat, high-sugar, and high-cholesterol diet combined with CCl4, which are similar to the changes in bile acids in MASH cases in clinical practice, suggesting that this model can be used to explore the interventional effect of drugs on the bile acid profile in MASH.
10.Mechanism of action of Homebox A6 in regulating the proliferation, invasion, metastasis, and apoptosis of HepG2 hepatoma cells
Yuting LIU ; Jingyin MAI ; Tianlu HOU ; Yang CHENG
Journal of Clinical Hepatology 2025;41(4):690-697
ObjectiveTo investigate the effect of Homebox A6 (HOXA6) on the proliferation, invasion, metastasis, and apoptosis of HepG2 hepatoma cells and its association with the PI3K/AKT signaling pathway. MethodsHepG2 hepatoma cells were cultured, and HOXA6 overexpression plasmid and siRNA were constructed and transfected into cells. The cells were randomly divided into empty plasmid group, HOXA 6 overexpression group, siRNA negative control group, and siRNA HOXA6 interference group. CCK8 assay was used to measure cell proliferation, Transwell assay was used to observe cell invasion, and wound healing assay was used to observe cell migration (related proteins TIMP3, MMP9, and MMP3). Flow cytometry was used to measure cell apoptosis (related proteins BAX and BCL2), the BCA method was used to measure protein concentration, and Western Blot was used to measure the expression of related proteins. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the SNK-q test was used for further comparison between two groups. ResultsCompared with the empty plasmid group, HOXA6 overexpression significantly promoted the proliferation, invasion, and migration of HepG2 hepatoma cells (all P<0.001), and there was a significant reduction in the protein expression of TIMP3 (P<0.001), while there were significant increases in the expression levels of MMP9 and MMP3 (both P<0.001). Compared with the siRNA negative control group, HOXA6 interference significantly inhibited the proliferation, invasion, and migration of HepG2 hepatoma cells (all P<0.001), and there was a significant increase in the protein expression of TIMP3 (P<0.001), while there were significant reductions in the expression levels of MMP9 and MMP3 (both P<0.001). Flow cytometry showed that compared with the empty plasmid group, HOXA6 overexpression inhibited the apoptosis of HepG2 hepatoma cells (P<0.001), with a significant reduction in the expression of the apoptosis-related protein BAX and a significant increase in the expression of BCL2 (both P<0.001). Compared with siRNA negative control group, HOXA6 interference promoted the apoptosis of HepG2 hepatoma cells (P<0.001), with a significant increase in the expression of BAX and a significant reduction in the expression of BCL2 (both P<0.001). Compared with the empty plasmid group, the HOXA6 overexpression group had significantly higher ratios of p-AKT/AKT and p-PI3K/PI3K (both P<0.001), and compared with the siRNA negative control group, the siRNA HOXA6 interference group had significantly lower ratios of p-AKT/AKT and p-PI3K/PI3K (both P<0.001). ConclusionHOXA6 can promote the proliferation, invasion, and metastasis of HepG2 hepatoma cells and inhibit their apoptosis by activating the PI3K/AKT signaling pathway through phosphorylation.

Result Analysis
Print
Save
E-mail