安络化纤丸对肝纤维化大鼠转化生长因子β1及相应信号通
路的影响
卢玮;高玉华;王珍子;蔡玉石;杨宇晴;苗玉麒;裴斐;刘学恩;庄辉
【期刊名称】《中华肝脏病杂志》 【年(卷),期】2017(025)004
【摘要】目的 中成药安络化纤丸已应用于临床治疗慢性乙型肝炎导致的肝纤维化,但其逆转肝纤维化的机制未明确.本研究利用四氯化碳(CCl4)诱导的大鼠肝纤维化模型,探索安络化纤丸抗纤维化的可能机制. 方法 27只雄性wistar大鼠随机分为对照组、模型组和治疗组(n=9),其中模型组和治疗组大鼠腹腔注射40％ CCl42rml/kg、对照组大鼠腹腔注射等渗盐水,每周2次、共6周.从建模第4周开始,治疗组大鼠每日灌胃安络化纤丸溶液(浓度为0.15 g/ml),其他两组大鼠灌胃等渗盐水,共6周.治疗3周和6周取血检测丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST).治疗6周结束后,取大鼠肝组织进行病理组织学评价,同时用荧光定量PCR法检测肝组织转化生长因子β1 (TGF β1)和Smads基因mRNA表达水平,用蛋白印迹法检测TGF β 1蛋白表达水平.计量资料是正态分布且满足方差齐性检验,多组间比较采用单因素方差分析;否则多组间比较采用非参数检验.等级资料多组间比较,采用非参数检验.结果 安络化纤丸治疗3周后,肝细胞损害减轻,治疗组大鼠ALT水平较模型组显著降低,(162.98±73.14) U/L对比(322.52±131.76)
U/L,P=0.047;AST
水平较模型组下降
39.8％,(537.56±306.06) U/L对比(892.98±358.19) U/L,P=0.053.安络化纤丸治疗6周后,治疗组大鼠肝纤维化程度较模型组有显著改善(P=0.002).治疗组大鼠TGF p 1基因mRNA和蛋白相对表达水平显著低于模型组(1.34±0.31对比
1.78±0.45,P=0.025;0.39±0.02对比0.57±0.04,P=0.003),差异均有统计学意义.结论 安络化纤丸能够逆转CCl4诱导的大鼠肝纤维化,其作用机制可能通过影响TGF β1产生,从而抑制肝星状细胞激活而发挥抗肝纤维化作用.%Objective The traditional Chinese medicine Anluohuaxianwan (ALHXW) has been used to treat liver fibrosis induced by chronic hepatitis B virus (HBV) infection.However,the anti-fibrosis mechanisms of ALHXW remain to be investigated.This study used a rat model of carbon tetrachloride (CCl4)-induced liver fibrosis to explore the potential antifibrogenic mechanisms of ALHXW.Methods Twentyseven male Wistar rats were randomly assigned to control group,model group,and treatment group (n =9 per group).Rats in the model and treatment group were injected intraperitoneally with 40％ CC14(2 ml/kg),and rats in the control group were administered saline twice a week for 6 weeks.Starting at week 4 following model construction,rats in the treatment group received daily gavages with ALHXW solution (concentration 0.15 g/ml) daily,while rats in the control and model groups were given saline for a total of 6 weeks.Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured from blood samples collected at the end of weeks 3,6 and 9.Histopathological examination of liver tissue was performed to evaluate liver fibrosis at week 9.At the same time,the mRNA expression of TGF-β1 and Smads in liver tissues was quantified by real-time reverse transcription polymerase chain reaction (RT-PCR),and TGF-
β1 protein level in the liver was measured by Western blot.Inter-group comparison was performed using analysis of variance (ANOVA) when the continuous data were normally distributed and satisfied the homogeneity
of
variance;otherwise,nonparametric
tests
were
used.Categorical data were compared between groups using nonparametric tests.Results ALHXW markedly alleviated liver injury in the treatment group after 3 weeks of therapy as indicated by a significantly reduced level of ALT compared with the model group [(162.98 ±73.14)U/L vs (322.52 ± 131.76)U/L,P =0.047],and a 39.8％ reduction in AST level compared with the model group[(537.56 ± 306.06)U/L vs (892.98 ± 358.19)U/L,P =0.053].Moreover,at the end of the 6-week therapy,histopathological diagnosis showed that liver fibrosis was significantly reduced in the ALHXWtreated group compared with that in the model group (P =0.002).The relative expression of TGF-β1 mRNA and protein in the liver were significantly lower in ALHXW-treated rats than that in model rats (1.34 ± 0.31 vs 1.78 ± 0.45,P =0.025;0.39 ± 0.02 vs 0.57 ± 0.04,P =0.003).Conclusion ALHXW treatment can reverse CCl4-induced liver fibrosis in rats.Its mechanisms of anti-fibrosis may occur through the inhibition of TGF-β1synthesis and TGF-β1/Smads signaling pathway,which in turn suppress the activation of hepatic stellate cells and thereby reverses fibrosis. 【总页数】6页(257-262)
安络化纤丸对肝纤维化大鼠转化生长因子β1及相应信号通路的影响
