Gan Shen Fu Fang ameliorates liver fibrosis in vitro and in vivo by inhibiting the inflammatory response and extracellular signalregulated kinase phosphorylation

BACKGROUND Liver fibrosis is a common health problem worldwide and there is still a lack of effective medicines.The Chinese herbal medicine,Gan Shen Fu Fang(GSFF)is composed of salvianolic acid B and diammonium glycyrrhizinate.In this study,we observed the effects of GSFF on liver fibrosis in vivo and in vitro in an attempt to provide some hope for the treatment.AIM To observe the effects of GSFF on liver fibrosis in vivo and in vitro and investigate the mechanism from the perspective of the inflammatory response and extracellular signal-regulated kinase(ERK)phosphorylation.METHODS Common bile duct-ligated rats were used for in vivo experiments.Hepatic stellate cells-T6(HSC-T6)cells were used for in vitro experiments.Hematoxylin and eosin staining and Masson staining,biochemical assays,hydroxyproline(Hyp)assays,enzyme-linked immunoasorbent assay and western blotting were performed to evaluate the degree of liver fibrosis,liver function,the inflammatory response and ERK phosphorylation.The CCK8 assay,immunofluorescence and western blotting were applied to test the effect of GSFF on HSC-T6 cell activation and determine whether GSFF had an effect on ERK phosphorylation in HSC-T6 cells.RESULTS GSFF improved liver function and inhibited liver fibrosis in common bile ductligated rats after 3 wk of treatment,as demonstrated by histological changes,hydroxyproline assays and collagen I concentrations.GSFF alleviated inflammatory cell infiltration and reduced the synthesis of pro-inflammatory cytokines[tumor necrosis factor-α(TNF-α)and interlukin-1β]and NF-κB.In addition,GSFF decreased ERK phosphorylation.In vitro,GSFF inhibited the viability of HSC-T6 cells with and without transforming growth factorβ1(TGF-β1)stimulation and decreased the synthesis of collagen I.GSFF had the greatest effect at a concentration of 0.5μmol/L.GSFF inhibited the expression ofα-smooth muscle actin(α-SMA),a marker of HSC activation,in HSC-T6 cells.Consistent with the in vivo results,GSFF also inhibited the phosphorylation of ERK and downregulated the expression of NF-κB.CONCLUSION GSFF inhibited liver fibrosis progression in vivo and HSC-T6 cell activation in vitro.These effects may be related to an alleviated inflammatory response and downregulated ERK phosphorylation.

The Protective Effect of Gan Shen Fu Fang on Liver Endothelial Cells in Common Bile Duct-ligated Rats

Objective: To elucidate the protective effect of Gan Shen Fu Fang(GSFF) on liver endothelial cells in common bile duct-ligated(CBDL) rats.Materials and Methods: Cirrhosis was induced by common bile duct ligation. The rats were divided into three groups: sham group, CBDL group, and GSFF group. After 2 weeks of ligation, rats in the GSFF group were administered GSFF. After 4 weeks, the hydroxyproline(Hyp)content of liver tissues was spectrophotometrically determined. The histological changes were evaluated by H and E and Masson staining.Transmission electron microscopy(TEM) and scanning electron microscopy(SEM) were used to observe the ultrastructural changes in the liver, especially in the liver sinusoidal endothelial cells(LSECs). Results: Hyp synthesis was significantly inhibited by GSFF, which agreed with the results from H and E and Masson staining for liver fibrosis. The TEM observations of CBDL rats revealed reduced hepatocyte microvilli and deposited fibrous tissue underneath LSECs. SEM confirmed the TEM findings and showed that the fenestrae of LSECs decreased and even disappeared in CBDL rats. The morphological results indicated hepatic sinusoid capillarization. GSFF promoted the restoration of fenestrae and reversed hepatic sinusoid capillarization. Conclusion: GSFF can inhibit Hyp synthesis, restore the fenestrae of LSECs, and reverse hepatic sinusoid capillarization in CBDL rats. These results provide a basis for future detailed investigations of the mechanism of action of GSFF in LSECs.

中药复方对DMN诱导的肝纤维化大鼠Ⅳ型胶原和TGF-β1蛋白表达的影响

目的:探讨中药复方对二甲基亚硝胺(DNM)诱导大鼠肝纤维化后Ⅳ型胶原和转化生长因子β1(TGF-β1)蛋白表达的影响,以了解其对肝纤维化的治疗作用和分子机制。方法:采用二甲基亚硝胺诱导大鼠肝纤维化模型,实验分为正常组、模型组和治疗组,观察肝组织羟脯氨酸含量,肝组织Ⅳ型胶原和转化生长因子β1(TGF-β1)蛋白表达水平及肝组织病理变化。结果:补肾柔肝方对肝纤维化有明显的治疗作用,治疗组羟脯氨酸含量要显著低于模型组,同时能使肝组织Ⅳ型胶原和转化生长因子β1(TGF-βl)蛋白表达水平下调。结论:补肾柔肝方可用于治疗肝纤维化。其机制可能与下调TGF-β1表达,抑制Ⅳ型胶原表达有关。

滋补肝肾方联合黄光对女性黄褐斑患者MASI评分的影响

目的 探讨滋补肝肾方联合黄光治疗女性黄褐斑患者的疗效。方法 选取2017年5月~2018年6月我院女性黄褐斑患者78例,按照随机数字表法分为研究组(n=39)和对照组(n=39)。对照组采用黄光治疗,研究组在对照组基础上采用滋补肝肾方治疗。比较两组疗效、治疗前后中医证候积分、黄褐斑面积及严重程度(MASI)评分、血清P物质(SP)、促黑激素(MSH)水平、皮肤病生活质量指数(DLQI)评分。结果 研究组总有效率92.31%(36/39)高于对照组71.79%(28/39,P<0.05);治疗3个月后研究组中医证候积分、MASI评分、DLQI评分低于对照组(P<0.05);研究组治疗3个月后血清SP、MSH水平低于对照组(P<0.05)。结论 滋补肝肾方联合黄光治疗女性黄褐斑患者效果显著,可改善症状,提高生活质量,降低血清SP、MSH水平。

甘肾方治疗腹痛并发症随机平行对照研究

[目的]观察甘肾方治疗腹痛并发症疗效。[方法]使用随机平行对照方法,将33例住院患者按病志号抽签简单随机分为两组。对照组33例东莨菪碱,0.5mg/次,1次/d,肌注;利多卡因,50~100mg/次,2次/d,静注;氯丙嗪,25~50mg/次,2次/d,静注。治疗组33例甘肾方（甘草、黄芪各25g,芍药、黄柏各5g,人参25g,升麻、葛根各15g,蔓荆子7.5g）,1剂/d,水煎200m L,早晚口服。连续治疗7d为1疗程。观测临床症状、腹痛、不良反应。治疗1疗程,判定疗效。[结果]治疗组痊愈17例,显效8例,有效6例,无效2例,总有效率93.94%。对照组痊愈16例,显效7例,有效4例,无效6例,总有效率81.82%。治疗组疗效优于对照组（P〈0.05）。[结论]甘肾方治疗腹痛并发症,疗效满意,无副作用,值得推广。