Prediction and validation of molecular biological mechanism of Fuzheng Huayu capsule in the treatment of liver cancer

Background:Fuzheng Huayu capsule(FZHY)combined with antiviral treatment has been shown to significantly reduce the risk of liver cancer in patients with hepatitis B cirrhosis.However,the potential of FZHY to directly treat liver cancer remains largely unknown.This study aims to investigate the molecular mechanism underlying the potential of FZHY in treating liver cancer.Methods:A network pharmacological analysis was performed using the Traditional Chinese Medicine Systems Pharmacology database to identify FZHY compounds and targets.Disease targets were searched using the Genecards database,and transcriptome data was downloaded from the NCBI database.Gene Ontology analysis was conducted using the DAVID database,and Kyoto Encyclopedia of Genes and Genomes analysis was based on KOBAS and bioinformatics methods.The Swissdock database was used for molecular docking.In cell experiments,the half inhibitory concentration(IC50)of FZHY was determined using the CCK8 method.The effects of FZHY on cell viability,apoptosis,and mitochondrial membrane potential were evaluated using a fluorescence microscope and flow cytometry.The molecular mechanism of FZHY in treating liver cancer was verified using quantitative polymerase chain reaction.Results:A total of 127 compounds and 184 proteins were identified as potential active ingredients and putative liver cancer-related targets.Additionally,1,899 liver cancer targets,279 transcriptome targets,and 3 pathways(p53 signaling pathway,apoptosis and PI3K-Akt pathway)were collected.The FZHY-targets-liver cancer interaction network was constructed.IC50 of FZHY lyophilized powder solution to liver cancer was 5.13 mg/mL(IC50=5.13 mg/mL).FZHY treatment led to an increase in the ratio of cell apoptosis and induced mitochondrial membrane potential damage,resulting in an increase in the number of dead cells.The expression levels of CCNB1 and BIRC5 were induced with FZHY treatment,while the expression levels of AKR1C3 and IGF2 were reduced.Conclusion:FZHY promotes apoptosis of liver cancer cells by acting on the p53 signaling pathway,apoptosis,and PI3K-Akt pathway.CCNB1,BIRC5,AKR1C3,and IGF2 are potential target proteins for FZHY in treating liver cancer.

Impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis

Background:Fuzheng Huayu tablet is a traditional Chinese medicine(TCM)used for the treatment of liver fibrosis and cirrhosis.However,whether the combination with Fuzheng Huayu tablet could affect the antiviral efflcacy of nucleos(t)ide remains a concern.The objective of this trial was to explore the impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis.Methods:A prospective,randomized control trial was conducted.Patients with compensated hepatitis B cirrhosis were randomly divided into the treatment group(entecavir capsule plus Fuzheng Huayu tablet)and the control group(entecavir capsule plus simulant of Fuzheng Huayu),and followed up for 48 weeks.The dynamic changes of HBV DNA load,the rate of serological conversion of HBeAg,liver function,renal function and liver stiffness measurement(LSM)were monitored.The general clinical data and adverse events were also recorded.Results:There was no significant difference in the rate of virological response and cumulative virological response between the treatment group and the control group(P>0.05).After 48 weeks of treatment,the HBeAg seroconversion rate,biochemical response rate and LSM value were 21.05%and 4.76%(P=0.164),86.96%and 65.96%(P=0.017),9.5 kpa and 10.6 kpa(P=0.827)in the treatment group and the control group,respectively.No serious adverse events related to the study therapy occurred during the trial.Conclusions:The antiviral entecavir combined with Fuzheng Huayu tablet did not affect the antiviral efflcacy of entecavir,but could improve the rate of biochemical response,and had a tendency to improve the rate of serological conversion of HBeAg and liver fibrosis in patients with hepatitis B cirrhosis.Fuzheng Huayu tablet is clinically safe for patients with hepatitis B cirrhosis.

Effects of Fuzheng Huayu 319 recipe on liver fibrosis in chronic hepatitis B

AIM To investigate clinic effects of Fuzheng Huayu 319 recipe (319 recipe) on liver fibrosis in chronic hepatitis B.METHODS Ninety-five patients with chronic hepatitis B were divide into the treated (63 cases) and control (32 cases) group, and orally administrated with 0.5g 319 capsule or 0.5g Dahuang Zhachong pill tid for 3 months, respectively. The liver functions and serological fibrotic markers were observed before and after treatment, 12 cases in the treated group were examined with liver biopsy.RESULTS Three hundreds nineteen recipe could remarkably decreased serum ALT level and total bilirubin and significantly improve serum albumin and A/G ratio. Its effects were better than Dahuang Zhachong pill. Before treatment, patients' serum monamine oxidase activities, tissue inhibitor of metalloproteinase (TIMP)-1, procollagen type Ⅲ and laminin were all higher than those of health peoples. These levels decreased remarkably after treatment, and urine hydroxyproline level increased significantly (P＜0.001-0.05). Compared with the control, the improvement in treated group was better than that in the control except TIMP-1. According to the scoring system for staging of chronic hepatitis, the fibrotic extents of 7 cases among 12 cases examined by liver biopsy decreased remarkably (1 case decreased by 3 scores, 5 by 2 scores, 1 by 1 score).CONCLUSION Fuzheng Huayu 319 recipe had good therapeutic effects on chronic hepatitis B, it could reverse the development of liver fibrosis to some extent. In general its effects were better than that of Dahuang Zhachong pill.

Fuzheng Huayu Capsule Improves Survival in Patients with HepatitisB-related Cirrhosis and Overt Hepatic Encephalopathy

Objective:Fuzheng Huayu(FZHY)capsules exert anti-liver fibrosis and cirrhosis effects.This study aimed to determine the effect of FZHY on the 6-month survival rate of patients with overt hepatic encephalopathy(OHE)and hepatitis B-related cirrhosis(HBC).Methods:A total of 517 patients with OHE and HBC treated between January 2008 and July 2018 were enrolled.Patients were divided into the FZHY(n=129)and control groups(n=388),and the 6-month survival rates were compared between the two groups.Propensity score matching(PSM)was used to reduce the bias caused by confounding factors.Results:In multivariable regression analysis,FZHY therapy was an independent protective factor of 6-month survival.After PSM(1:2),the FZHY and control group comprised 126 and 252 patients,respectively.The 6-month survival rate was remarkably higher in the FZHY group than in the control group(P<0.005).FZHY users,especially those with a model for end-stage liver disease score>20 and Child-Pugh class C,benefited significantly from FZHY therapy.Conclusions:Adjuvant therapy with FZHY may be associated with improved survival in patients with OHE and HBC.However,further clinical studies are required to validate our findings.

Beneficial Effects of Traditional Chinese Medicine Fuzheng Huayu on the Occurrence of Hepatocellular Carcinoma in Patients with Compensated Chronic Hepatitis B Cirrhosis Receiving Entecavir:A Multicenter Retrospective Cohort Study

Background and Aims:The application of antifibrotic drugs to treat patients with chronic liver diseases who are receiving antiviral therapies for hepatocellular carcinoma(HCC)has not been established.Here,we aimed to assess the impact of the Traditional Chinese Medicine Fuzheng Huayu(FZHY)on the occurrence of HCC in patients with hepatitis B virus-related compensated cirrhosis receiving the antiviral drug entecavir(ETV).Methods:A multicenter retrospective cohort study was performed.Compensated liver cirrhosis patients were divided into the ETV+FZHY group or the ETV group according to treatment.The cumulative incidence of HCC was analyzed using Kaplan-Meier and log-rank tests.Propensity score matching was used for confounding factors.Stratified analysis and Cox regression were used to determine the effects of FZHY on the occurrence of HCC and liver function decompensation.Results:Out of 910 chronic hepatitis B patients,458 were in the ETV+FZHY group and 452 were in the ETV group.After propensity score matching,the 5-year cumulative incidence of HCC was 9.8%in the ETV+FZHY group and 21.8%in the ETV group(p<0.01).The adjusted hazard ratio for HCC was 0.216(0.108,0.432)when FZHY treatment was>36 months.Age,diabetes,alanine aminotransferase,γ-glutamyl transpeptidase,albumin,hepatitis B e-antigen,and fibrosis 4 score were associated with the occurrence of HCC.FZHY decreased the risk of HCC in patients aged>45 years with a hepatitis B virus DNA level of≥2,000 IU/l.Conclusion:Adjunctive FZHY treatment reduced HCC occurrence in patients with hepatitis B virus cirrhosis who were treated with ETV,possibly due to the antifibrotic properties of FZHY.

Fuzheng Huayu preparation(扶正化瘀胶囊/片)combined with tenofovir disoproxil fumarate on hepatitis B:a systematic review and Meta-analysis

OBJECTIVE:To systematically evaluate the effectiveness of Fuzheng Huayu preparation(扶正化瘀胶囊/片,FZHY)plus tenofovir disoproxil fumarate(TDF)on hepatitis B.METHODS:Numerous databases—PubM ed,Embase,Cochrane Library,China National Knowledge Infrastructure Database,WanFang Database,China Science and Technology Journal Database,and China Biological Medicine Database—were searched to identify the randomized controlled trials published from the inception of the database to November 2021.Two researchers independently conducted literature screening,data extraction,and bias risk assessment.Rev Man 5.4 software was used for Meta-analysis.RESULTS:Eight studies involving 990 patients met the inclusion criteria in the current Meta-analysis.Levels of alanine transaminase,aspartate aminotransferase,total bilirubin,hyaluronic acid,typeⅢprocollagen,laminin,and type IV collagen after combination therapy were significantly lower than those after TDF therapy alone.However,albumin levels did not differ significantly between the two regimens.Subgroup analysis based on disease progression suggested that the combination therapy improved albumin levels in patients with chronic hepatitis B but not in patients with hepatitis B-related cirrhosis.Moreover,subgroup analysis based on treatment duration suggested that the albumin levels were increased and the typeⅢprocollagen levels were decreased with the>24-week combination therapy but not with the≤24-week combination therapy.CONCLUSIONS:A combination regimen of TDF and FZHY is more effective in treating hepatitis B than TDF alone.The combination therapy can effectively alleviate hepatic fibrosis and improve liver function.However,more standardized,highquality studies with larger sample sizes are warranted to validate the study results.

Fuzheng Huayu Capsule(扶正化瘀胶囊)in the Treatment of Liver Fibrosis:Clinical Evidence and Mechanism of Action

Liver fibrosis represents the wound healing response to liver injury from a wide variety of etiologies. Remarkable progresses have been shown in the field of liver fibrosis in a range of areas in the past years. In particular, the reversibility of liver fibrosis has been well documented in both patients and animal models. Great progresses have been made in the treatment of liver fibrosis with Chinese medicine. This review summarizes the effects of Fuzheng Huayu Capsule （扶正化瘀胶囊, FZHYC） in treating liver fibrosis and inflammation induced by chronic hepatitis B in clinical trials and the mechanism of action of FZHYC in reversing liver fibrosis in vivo and in vitro experiments.

Treatment of Posthepatitic Cirrhosis by Fuzheng Huayu Tablet(扶正化瘀片) for Reinforcing Qi and Resolving Stasis

Objective： To investigate the efficacy and safety of the Fuzheng Huayu Tablet （扶正化瘀片, FZHYT）, which is used to reinforce qi and resolve stasis in patients with posthepatitic cirrhosis （PHC）. Methods： A multicenter, randomized, controlled clinical trial was conducted in 180 patients with PHC. The patients were randomly assigned using random numbers to a treatment group treated with FZHYT and a placebo group; the treatment course was 6 months for both groups. Overall response, adverse events （AEs）, and the 2-year survival rate were assessed after treatment. Evaluations were made on changes in liver function, liver fibrosis, coagulation, hemodynamics, degrees of esophagogastric varices, ascites, quality of life （QOL）, and scores of main symptoms. Results： The overall response was significantly higher in the treatment group than the placebo group （86.7% vs. 62.2%, P〈0.01）. Patients in both groups had significant improvements in liver function [total bilirubin （TBIL）, albumin （ALB）], liver fibrosis [hyaluronic acid （HA）, type 1V collagen （CIV）], coagulation [prothrombin time （PT）, activated partial thromboplastin time （APTT）, fibrinogen （FIB）, and thrombin time （TT）], hemodynamics portal venous flow （PVF）, and splenic vein flow （SVF） after treatment. Between-group comparisons showed that compared with the placebo group patients in the treatment group achieved significantly greater improvements in TBIL, ALB, HA, C IV, PT, AP＇I-r, PVF, SVF, time to ascites resolution, 2-year survival, QOL, and symptom scores （P〈0.05 or P〈0.01）. There were no significant AEs during the treatment. Conclusion： FZHYT is effective and safe for the treatment of hepatic cirrhosis as it is associated with improved liver function, liver fibrosis, coagulation, portal hypertension state, QOL, 2-year survival rate, and fewer AEs.

Fuzheng Huayu Formula(扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）activity.Methods：Thirty-two Sprague-Dawley rats were randomly divided into 3 groups：normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline（Hyp）and collagen deposition in kidney were observed.The activities of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）,the content of glutathione（GSH）and malondialdehyde（MDA）of kidney were tested.The expressions of inhibitor-κappa B（IκB）,phospho-IκB（p-IκB）,tumor necrosis factor-α（TNF-α）,matrix metalloproteinase-2（MMP-2）andα-smooth muscle actin（α-SMA）were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results：Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney（P〈0.01）.FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats（P〈0.01）.GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats（P〈0.01）.FZHY significantly increased GSH content and decreased MDA content in the model rats（P〈0.01）.The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression（P〈0.01）.The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group（P〈0.01）,FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions（P〈0.01）.Conclusions：FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.

Histological Outcome of Fuzheng Huayu plus Entecavir Combination Therapy in Chronic Hepatitis B Patients with Significant Liver Fibrosis

Publications>Journals>Journal of Clinical and Translational Hepatology>Article Full Text ORIGINAL ARTICLE OPEN ACCESS Histological Outcome of Fuzheng Huayu plus Entecavir Combination Therapy in Chronic Hepatitis B Patients with Significant Liver Fibrosis Hong-Lian Gui#,1,Chang-Qing Zhao#,2,Yan Wang3,Hong-Tu Gu2,Wei-Jing Wang1,4,Wei Cai1,Qing Guo1,Shi-San Bao5,Lie-Ming Xu*,2 and Qing Xie*,1 Author information Journal of Clinical and Translational Hepatology 2020;8(3):277-284DOI:10.14218/JCTH.2020.00004 Abstract Background and Aims:To evaluate the efficacy of Fuzheng Huayu(FZHY),a Chinese herbal formula,plus entecavir(ETV)in regression of liver fibrosis in chronic hepatitis B(CHB)patients with significant fibrosis/cirrhosis.Methods:The current study was a two-center,randomized,double-blind and placebo-controlled pilot study.Fifty-two currently untreated chronic hepatitis B patients with Ishak fibrosis score≥3 points were identified and 1:1 randomized into FZHY plus ETV combination and placebo plus ETV groups.The second liver biopsy was performed after 48-week treatment.Necroinflammatory improvement and regression of fibrosis were assessed.Fine changes in different collagen features in paired liver biopsies were evaluated by dual-photon microscopy for both groups.Results:Forty-nine patients completed the full course of treatment;forty-six of them underwent second liver biopsy(for which twenty-two were in the combination group and twenty-four were in the control group).Compared to those in the control group,patients in the combination group had significantly higher rate of fibrosis regression(82%vs.54%)(p<0.05).Furthermore,the necroinflammatory improvement was greater in the combination group than in the control group(59%vs.25%,p<0.05).Among the more than 80 collagen parameters in the dual-photon analysis,5 decreased significantly in the combination group compared to the control group(p<0.05).However,no significant improvement was detected in either biochemical,virologic or serologic responses between these two groups at week 48.Conclusions:The combination therapy of FZHY plus ETV for 48 weeks resulted in a higher rate of necroinflammatory improvement and fibrosis regression than ETV alone in chronic hepatitis B patients with significant fibrosis/cirrhosis.The clinical trial number is ChiCTR-TRC-11001377.

Fuzheng Huayu recipe, a traditional Chinese compound herbal medicine, attenuates renal interstitial fibrosis via targeting the miR-21/PTEN/AKT axis

Objective: MicroRNAs(miRNAs) may be viable targets for treating renal interstitial fibrosis(RIF). Fuzheng Huayu recipe(FZHY), a traditional Chinese compound herbal medicine, is often used in China to treat fibrosis. This study sought to assess the mechanisms through which FZHY influences miRNAs to treat RIF.Methods: RIF was induced in rats by mercury chloride and treated with FZHY. Hydroxyproline content,Masson’s staining and type I collagen expression were used to evaluate renal collagen deposition.Renal miRNA profiles were evaluated using a miRNA microarray. Those miRNAs that were differentially expressed following FZHY treatment were identified and subjected to bioinformatic analyses. The miR-21 target gene phosphatase and tensin homolog(PTEN) expression and AKT phosphorylation in kidney tissues were assessed via Western blotting. In addition, HK-2 human proximal tubule epithelial cells were treated using angiotensin II(Ang-II) to induce epithelial-to-mesenchymal transition(EMT), followed by FZHY exposure. miR-21 and PTEN expressions were evaluated via quantitative reverse transcriptionpolymerase chain reaction(qRT-PCR), while E-cadherin and a-smooth muscle actin(a-SMA) expressions were assessed by immunofluorescent staining and qRT-PCR. Western blotting was used to assess PTEN and AKT phosphorylation.Results: FZHY significantly decreased kidney collagen deposition, hydroxyproline content and type I collagen level. The miRNA microarray identified 20 miRNAs that were differentially expressed in response to FZHY treatment. Subsequent bioinformatic analyses found that miR-21 was the key fibrosis-related miRNA regulated by FZHY. FZHY also decreased PTEN expression and AKT phosphorylation in fibrotic kidneys. Results from in vitro tests also suggested that FZHY promoted E-cadherin upregulation and inhibited a-SMA expression in Ang-II-treated HK-2 cells, effectively reversing Ang-II-mediated EMT. We also determined that FZHY reduced miR-21 expression, increased PTEN expression and decreased AKT phosphorylation in these cells.Conclusion: miR-21 is the key fibrosis-related miRNA regulated by FZHY. The ability of FZHY to modulate miR-21/PTEN/AKT signaling may be a viable approach for treating RIF.

Treatment of HBV Cirrhosis with Fuzheng Huayu Tablet(扶正化瘀片)and Entecavir:Design of a Randomized,Double-Blind,Parallel and Multicenter Clinical Trial

Background Antiviral therapy can lead to regression of fibrosis in chronic hepatitis B(CHB),but it has a limited effect on cirrhosis.Chinese medicines(CMs),particularly Fuzheng Huayu Tablet(扶正化瘀片,FZHY),have an antifibrotic effect in patients with CHB.Objective To observe the safety and efficacy of adjunctive FZHY in patients with hepatitis B virus(HBV)cirrhosis,this study was designed as a randomized,placebo-controlled,double-blind,parallel assignment,multicenter trial at 20 centers in China.The total 700 naive patients will be enrolled with compensate cirrhosis due to HBV,and randomly assigned into 2 groups,receiving entecavir(0.5 mg,daily)and FZHY placebo(1.6 g,3 times a day),or entecavir(0.5 mg,daily)and FZHY(1.6 g,3 times a day),respectively.The primary endpoint was histological improvement at week 48.The secondary outcome is the decline values of liver fibrosis using the noninvasive methods from baseline to week 48 in each arm of the study.Adverse events such as stomach upset,headache,fatigue,dizziness,nausea will be strictly recorded.Discussion Through this study,we hope to generate a solid evidence for the therapeutic strategy of HBV cirrhosis with a combination of anti-viral such as ETV and anti-fibrotic herbal product such as FZHY.Protocol version:Version 1.3,Date:2014.12.4.Trial registration number:NCT02241590.

Influence of Fuzheng Huayu Tablet(扶正化瘀片) on Mental State and Social Function of Patients with Post-Hepatitis B Liver Cirrhosis

Objective： To observe the influence of Fuzheng Huayu Tablet （扶正化瘀片, FZHYT） on mental state and social activity of patients with post-hepatitis B liver cirrhosis （LC-HB）. Methods： Adopting grouped randomized double-blinded control method, 180 LC-HB patients in 3 research centers were distributed to 2 groups, the treated group and the control group, 90 in each group. Patients in the treated group were administered with FZHYT; while those in the control group treated with conventional therapy combined with placebo, the course for all patients were 6 months. Their mental state and social activity were evaluated before treatment, after 3 months＇ treatment and at terminal of the 6-month therapeutic course by estimating with Zung self-rating anxiety scale （SAS）, self-rating depression scale （SDS） and social deficit screening scale （SDSS）. Additionally, the basic demographic materials, liver function, cirrhosis index, hepatic and splenic images, blood coagulation function, etc. in the patients were tested and compared as well. Results： As compared with before treatment, the normal rate of SAS and SDS scores increased and the social deficit rate decreased in the treated group significantly after treatment, showing statistical significance （P〈0.05 or P〈0.01）; while in the control group, change was only shown in the social deficit （P〈0.01）, inter-group comparisons after treatment showed significant differences in all the three indexes （P〈0.05 or P〈0.01）. Additionally, after treatment, levels of liver function, cirrhosis, blood coagulation function and splenomegaly in the treated group were all improved significantly （P〈0.05 or P〈0.01）, and the improvements were better than those in the control group （P〈0.01） in levels of total bilirubin （TBIL）, albumin （ALB）, type 1V collagen （1V-C）, prothrombin time （PT）, prothrombin activity （PTA）. Conclusion： Most patients of LC-HB have mental disturbance and social activity deficit, which could definitely be improved by intervention with Chinese FZHYT.

Active Components Formulation Developed from Fuzheng Huayu Recipe for Anti-Liver Fibrosis

Objective:To screen the active components from Fuzheng Huayu Recipe(FZHY)and redesign a new recipe composed of the active components,and validate the effect of active components formulation from FZHY against liver fibrosis.Methods:Thirty-two components from FZHY were evaluated for their activities against liver fibrosis respectively,with 6 kinds of cell models in vitro,including oxidative stressed hepatocyte in L-02,hypoxia injured/proliferative hepatic sinusoidal endothelial cells in SK-HEP-1 and human hepatic sinusoidal endothelial cells(HHSEC),and activated hepatic stellate cell in LX-2.The comprehensive activity of each component against liver fibrosis was scored according to the role of original herbs in FZHY and cell functions in fibrogenesis.Totally 7 active components were selected and combined with equal proportion to form a novel active components formulation(ACF).The efficacy of ACF on liver fibrosis were evaluated on activation of LX-2 and proliferation of HHSEC in vitro and in liver fibrosis model mice induced by dimethylnitrosamine(DMN).Totally 72 mice were divided into6 groups using a random number table,including normal,high-dose ACF control(20μmol/L×7 components/kg body weight),model,low-,medium-,high-dose ACF groups(5,10,20μmol/L×7 components/kg body weight,respectively).Hematoxylin eosin and Sirius red stainings were used to observe inflammation and fibrosis change of liver tissue;scanning electron microscopy(SEM)and transmission electron microscopy(TEM)were utilized to observe the effect of ACF on ultrastructure of hepatic sinusoids.Results:Fifteen components from FZHY showed higher scores for their activity on against liver fibrosis.Among them,7 components including tanshinoneⅡA,salvianolic acid B,cordycepin,amygdalin,quercetin,protopanaxatriol,and schizandrin B were recombined with equal proportions to form ACF.ACF at 1,2,4μmol/L showed strong inhibitory effects on activation of LX-2 and proliferation of HHSEC in vitro(all P<0.01).Compared with the model group,ACF attenuated liver collagen deposition,improved sinusoidal capillarization in a dose-dependent manner(all P<0.05).Conclusions:ACF exerts a satisfactory effect against experimental liver fibrosis and attenuates sinusoidal capillarization,which warrant a further research and development for herbal components formulation on liver fibrosis.

Serum Pharmacological Effects of Fuzheng Huayu Decoction(扶正化瘀方) on lto Cell Proliferation and Collagen Synthesis in Rats

Objective: To explore mechanism of the effect of Fuzheng Huayu Decoction on Ito cell proliferation and collagen synthesis. Methods: Biological effects of the drug serum taken from rats fed with Fuzheng Huayu decoction （FZHYD） on cell proliferation and collagen synthesis of Ito cell line were observed. Serum collected from rats at various times （1, 2 or 3 hours） after animals were fed with FZHYD once （It） or twice （2t,2nd time repeated with the same dose）, was incubated with Ito cell. Results: The 1t-2h serum in O.46 g/kg FZHYD group, it seems to prevent cell proliferation, but had no significant difference. While all 2t drug serum could inhibit cell proliferation. The 2t-1h drug serum had no obvious effects on Ito cell morphology, but could improve cell viability, its effect on cell proliferation was dose and serum concentration dependent. Also the 2t-1h drug serum in all doses could inhibit Ito cells both intracellular and extracellular collagen production, the drug serum of 0.46 g/kg group had inhibitory effect in serum concentration dependent manner and similar effect as colchicine. Conclusion: Inhibition of Ito cells proliferation and collagen synthesis is perhaps One of main mechanism of FZHYD in antifibrotic action.

Effects of Fuzheng Huayu Decoction on plasma proteome in cirrhosis:preliminary experimental study with rats

The aim of this paper is to study the effects of Fuzheng Huayu Decoction on the plasma proteome in cirrhotic rats.Twenty-six male Sprague-Dawley(SD)rats were randomly divided into three groups:cirrhotic model group(n510),treated with CCl_(4)(CCl_(4)/olive oil:v/v51:1);Fuzheng Huayu Decoction intervention group(n510),treated with CCl_(4)+Fuzheng Huayu Decoction;and normal control group(n56),treated with olive oil only.After 8 weeks,blood samples were collected from the inferior vena cava to undergo bi-dimensional electrophoresis(2DE)and analysis by PDQuest 7.3 software.Differential protein spots were cut,enzyme hydrolysis was conducted,and peptide fragments extracted from the mixture underwent mass spectrometry(MS)with MALDI-TOF-TOF-MS.The liver fibrogenesis was assessed using a digital image analysis instrument of Masson’s trichrome stained sections.The fibrosis area of the Fuzheng Huayu Decoction was(8.9±3.7)%,significantly smaller than that of the cirrhotic model group[(12.4±4.7)%,P<0.05].Ten markedly changed protein spots were identified by MALDI-TOFTOF-MS.Eight of the 10 proteins,including plasma glutathione peroxidase,plasma glutathione peroxidase precursor,prealbumin,haptoglobin,apolipoprotein A-IV precursor,complement C4,inter-alpha-inhibitor H4 heavy chain,and serine/threonine-protein kinase microtubuleaffinity regulating kinase 1(MARK1)were expressed very lowly in the cirrhotic model group while they were expressed highly in the Fuzheng Huayu Decoction group.The expression of liver regeneration-related protein LRRG03 and vimentin increased in the cirrhotic model group,and reduced in the FuzhengHuayu Decoction group.Some proteins related to oxidative stress,cell proliferation and transformation have changed in the plasma of cirrhosis induced by CCl_(4).Fuzheng Huayu Decoction promotes protein synthesis and plays an anti-fibrotic role by antioxidation and accommodation of cell proliferation and transformation.

Seropharmalogical Effects of Fuzheng Huayu decoction on rat Ito cell morphology and functions in culture

IM To investigate the mechanisms of antiliver fibrosis actions of Fuzheng Huayu (FZHY, strengthen the body resistance and promote blood circulation) decoction.METHODS Ito cells were isolated from rats and cultured, the sera were collected from the normal rats after administration of FZHY decoction and added to the subcultured cells. Ito cell morphology was observed under the contrast microscopy, cells viability was measured by Pro incorporation, proliferation by TdR incorporation and MTT colorimetric assay, collagen synthesis rate by Pro incorporation and collagenase digestion. The effects of FZHY decoction on Ito cell morphology and functions were observed.RESULTS The rat serum containing FZHY decoction had no influence on Ito cell morphology, but could improve the cell viability. It could markedly inhibit Ito cell proliferation and collagen synthesis, these effects tend to be drug dose and drug sera concentration dependent.CONCLUSION Seropharmalogical method can be efficiently used to investigate the pharmacological mechanism of antifibrotic traditional Chinese herbs and formulae. The inhibition of Ito cell proliferation and collagen synthesis may be one of the major mechanisms of the antifibrosis actions in FZHY decoction.

扶正化瘀胶囊对肝纤维化发展的影响

目的探讨扶正化瘀胶囊(FZHY)对肝纤维化发展的影响。方法CCK-8法检测FZHY对LX-2细胞活性的影响。RT-qPCR和Western blot法测定LX-2转染TGF-β或/和给予FZHY后α-SMA水平。选取2022年1月至2022年12月在上海第九人民医院接受抗病毒治疗的慢性乙型肝炎患者80例作为研究对象,分为对照组和治疗组,治疗组每日口服扶正化瘀胶囊1.5 g,一日3次,瞬时弹性成像检查各组治疗前后瞬时肝弹性F值。结果FZHY明显抑制LX-2生长,并与剂量呈正相关(P<0.05)。过表达TGF-β后α-SMA mRNA明显增加(57.14±4.81),与对照组相比差异显著(P<0.001)。同时予10%或20%FZHY时,α-SMA mRNA不同程度降低(37.42±3.65及21.79±0.496),与TGF-β组相比,α-SMA mRNA降低差异有统计学意义(P<0.05,P<0.001)。Western blot显示,FZHY能部分抵消TGF-β引起的α-SMA表达上调。治疗组瞬时肝弹性F值较干预前有所下降,但差异无统计学意义(P>0.05)。结论FZHY可通过影响LX-2细胞增殖、调节α-SMA表达来实现抗纤维化作用。

扶正化瘀散结法联合西药治疗厌食-恶病质综合征有效性的Meta分析

目的系统评价扶正化瘀散结法联合西药治疗厌食-恶病质综合征的有效性。方法系统检索维普(VIP)、中国知网(CNKI)、万方数据知识服务平台(Wan Fang)、中国生物医学文献数据库(CBM)、PubMed、Web of Science、Embase数据库、Cochrane Library数据库,利用Jadad、Cochrane协作网RCT质量评价标准对纳入文献的研究质量进行评价,Meta分析提取数据。结果1)共纳入17个随机对照研究(RCT),患者1423例,纳入研究质量一般;2)Meta分析合并RR值包括体重变化[RR=1.36,95%CI(1.08,1.71),P=0.009]、KPS评分变化[RR=1.44,95%CI(1.29,1.60),P<0.00001]、进食量变化[RR=1.33,95%CI(1.15,1.55),P=0.0002]、白蛋白变化[MD=3.01,95%CI(2.10,3.93),P<0.00001]以及血红蛋白变化[MD=10.74,95%CI(1.91,19.58),P=0.02]。结论本研究证实扶正化瘀散结法在厌食-恶病质综合征应用的有效性优于单纯西药治疗或结合营养支持治疗。

扶正化瘀片联合多烯磷脂酰胆碱治疗慢性乙肝肝硬化患者的疗效

目的:探讨扶正化瘀片联合多烯磷脂酰胆碱治疗慢性乙肝肝硬化患者的疗效及其对肝功能、肝纤维化和血清指标的影响。方法:选取106例慢性乙肝肝硬化患者,按照治疗方式不同分为对照组和研究组,每组各53例。对照组给予常规治疗和多烯磷脂酰胆碱治疗;研究组在对照组基础上给予扶正化瘀片治疗,两组均治疗24周。比较两组患者的疗效、肝功能、肝纤维化指标、血清指标水平及不良反应发生情况。结果:研究组的临床总有效率为96.23%,高于对照组的83.02%(P<0.05)。治疗后,两组患者的总胆红素(TBIL)、丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、透明质酸酶(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(IV-C)、层粘连蛋白(LN)、D二聚体(D-D)、总胆汁酸(TBA)水平均降低(P<0.05),且研究组均低于对照组(P<0.05);两组患者的白蛋白(ALB)、胆碱酯酶(CHE)水平均升高(P<0.05),且研究组高于对照组(P<0.05),两组不良反应总发生率无统计学差异(P>0.05)。结论:扶正化瘀片联合多烯磷脂酰胆碱治疗慢性乙肝肝硬化患者疗效显著,可有效改善患者肝功能,减轻肝纤维化,调节D-D、TBA及CHE的表达水平,并具有较好的安全性。