文献类型：期刊文献

中文题名：舒肝和络醒脾方对肝纤维化模型大鼠肝组织Wnt/β-Catenin信号通路的影响

英文题名：Shugan Heluo Xingpi Decoction inhibits liver fibrosis through regulation of Wnt/β-Catenin signaling pathway

作者：王奎淞[1];张秋菊[2];魏书瑶[3];殷世鹏[4];李婕妠[1];陈世玉[1];郭嘉琪[1];赵鲲鹏[1]

第一作者：王奎淞

机构：[1]甘肃中医药大学中医临床学院,兰州730000;[2]甘肃中医药大学基础医学院,兰州730000;[3]甘肃中医药大学公共卫生学院,兰州730000;[4]甘肃中医药大学图书馆古籍部,兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2022

卷号：38

期号：10

起止页码：2265

中文期刊名：临床肝胆病杂志

外文期刊名：Journal of Clinical Hepatology

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；

基金：国家自然科学基金(81660772);甘肃省高等学校创新基金项目(2020A-074);甘肃中医药大学科研创新基金(2018-03)。

语种：中文

中文关键词：肝硬化;Wnt信号通路;舒肝和络醒脾方

外文关键词：Liver Cirrhosis;Wnt Signaling Pathway;Shugan Heluo Xingpi Decoction

摘要：目的观察舒肝和络醒脾方对CCl_(4)诱导的肝纤维化模型大鼠的治疗作用,并基于Wnt/β-Catenin信号通路探讨其作用机制。方法将60只雄性SPF级Wistar大鼠随机分为空白对照组,模型组,阳性对照组,舒肝和络醒脾方高、中、低剂量组,每组10只,模型和用药大鼠给予腹腔注射40%CCl_(4)油溶液诱导肝纤维化模型。空白对照组和模型组给予10 mL/kg生理盐水灌胃,阳性对照组用水飞蓟宾葡甲胺溶液50 mg/kg灌胃,舒肝和络醒脾方高、中、低剂量组分别按12.42 g/kg、6.21 g/kg和3.11 g/kg(生药/体质量)灌胃,每日1次,持续8周。给药8周后处死,期间动态监测各组大鼠的生理状况。处死后取血清于碱水解比色法检测羟脯氨酸,于全自动生化分析仪检测AST、ALT、总蛋白和Alb水平;HE染色及Masson染色观察肝脏的病理学形态改变;RT-qPCR法、Western Blot法检测Wnt1、β-Catenin和Cyclin D1的mRNA和蛋白水平的表达。计量资料多组间比较采用one-way ANOVA分析,进一步两两比较采用LSD-t检验。结果与模型组比较,飞蓟宾葡甲胺溶液和舒肝和络醒脾方用药干预后的大鼠生理状况改善明显;血液学指标方面,与模型组比较,阳性对照组和舒肝和络醒脾方干预后大鼠血清中HYP、ALT、AST和Glo水平显著降低(P值均<0.05),总蛋白、Alb和A/G水平明显升高(P值均<0.05);与阳性对照组比较,舒肝和络醒脾方低剂量组的ALT、AST水平显著升高(P值均<0.05),总蛋白、Alb和A/G水平明显降低(P值均<0.05);肝组织学方面,HE染色显示阳性对照组可见门静脉轻度纤维化伴少量纤维间隔,肝细胞轻微脂肪变;舒肝和络醒脾方低剂量组可见明显门静脉纤维化伴少量纤维间隔;舒肝和络醒脾方中剂量组可见少量门静脉轻微纤维化;舒肝和络醒脾方高剂量组未观察到明显异常。Masson染色显示,舒肝和洛醒脾方高剂量组在胶原沉积的治疗效果方面优于水飞蓟宾葡甲胺组和舒肝和络醒脾方中、低剂量组(P值均<0.05),与舒肝和络醒脾方高剂量组基本等效。水飞蓟宾葡甲胺和舒肝和络醒脾方高、中剂量干预对Wnt1、β-Catenin和Cyclin D1的mRNA水平和蛋白表达的抑制作用更明显(P值均<0.05)。结论舒肝和络醒脾方能够发挥稳定的抗肝纤维化作用,高剂量作用效果较为显著,调节Wnt/β-Catenin信号通路可能为其抗肝纤维化的信号途径之一。
Objective To investigate the therapeutic effect of Shugan Heluo Xingpi Decoction on CCl_(4)-induced liver fibrosis in rats,and the underlying molecular mechanism.Methods Sixty male SPF Wistar rats were randomly assigned to six groups:blank control,model,positive control,high,medium or low dose groups of Shugan Heluo Xingpi Decoction(n=10 per group).The liver fibrosis rat model was induced by an intraperitoneal injection of 40% CCl_(4) oil solution.Rats in the blank control and model groups were administered 10 mL/kg normal saline by gavage,rats in the positive control group were administered 50 mg/kg silibinin meglumine by gavage,while rats in high-dose,medium-dose and low-dose groups of Shugan Heluo Xingpi Decoction were administered 12.42 g/kg,6.21 g/kg and 3.11 g/kg(crude drug/body weight)by gavage,respectively,daily for 8 weeks.Rats was sacrificed after 8 weeks,during which the physiological status of rats in each group was dynamically monitored.Following sacrifice,serum was collected to detect HYP using alkaline hydrolysis colorimetry and the expression levels of AST,ALT,total protein(TP),and Alb using an automatic biochemical analyzer.The pathological morphological changes in the liver were detected by H&E staining and Masson staining,and the mRNA and protein levels of Wnt1,β-Catenin and Cyclin D1 were detected by RT-qPCR and Western Blot.Measurement data were compared across groups using one-way ANOVA with post-hoc LSD-t test.Results Compared with the model group,after silibinin meglumine and Shugan Heluo Xingpi Decoction intervention,the physiological status of rats was significantly improved;serum levels of HYP,ALT,AST and Glo were significantly decreased,while serum levels of TP,Alb and A/G were significantly increased(all P<0.05).Compared with the positive control group,serum levels of ALT and AST were significantly increased(all P<0.05),while the levels of TP,Alb and A/G were significantly decreased(all P<0.05)in low-dose group of Shugan Heluo Xingpi Decoction.H&E staining showed mild portal vein fibrosis with a few fibrous septa and mild steatosis of hepatocytes in the positive control group,obvious portal vein fibrosis with a few fiber septum in the low dose group,a few portal vein fibrosis in the medium dose group,while no obvious abnormality in the high dose group of Shugan Heluo Xingpi Decoction.Masson staining revealed that the therapeutic effect of high dose group of Shugan Heluo Xingpi Decoction on collagen deposition was superior to silibinin meglumine and medium and low dose of Shugan Heluo Xingpi Decoction(all P<0.05),and was generally equivalent to high dose of Shugan Heluo Xingpi Decoction.Silibinin meglumine and medium and high doses of Shugan Heluo Xingpi Decoction inhibited more significantly the mRNA and protein expression of Wnt1,β-catenin and Cyclin D1(all P<0.05).Conclusion Shugan Heluo Xingpi Decoction shows anti-hepatic fibrosis effect,with a greater effect at higher doses.Regulating Wnt/β-Catenin signaling pathway may be one of the underlying molecular mechanisms.