摘要
目的观察疏肝健脾方对肝纤维化大鼠自噬蛋白Beclin-1和LC3-Ⅱ的影响。方法将60只SD大鼠,随机分为正常组、模型组、疏肝健脾方(1.5、3.0、6.0 g/kg)组和阳性药秋水仙碱(0.1×10-3g/kg)组,每组10只。除正常组外,其余各组采用50%四氯化碳(CCl4)每周2次,连续12周皮下注射的方法,诱导肝纤维化大鼠模型。造模后第7周,灌胃给予不同剂量的疏肝健脾方和秋水仙碱,正常组和模型组给予等量溶媒,每天1次,连续6周。HE和Masson染色观察肝脏病理组织学损伤程度,免疫组化法检测肝组织中Ⅰ型胶原(CollagenⅠ)、α平滑肌肌动蛋白(α-SMA)、Beclin-1、LC3-Ⅱ蛋白的表达。结果与正常组比较,模型组大鼠肝脏病理组织学损伤程度明显加重,肝组织中CollagenⅠ、α-SMA、Beclin-1和LC3-Ⅱ蛋白表达水平显著升高(P<0.01);与模型组比较,疏肝健脾方(3.0、6.0 g/kg)组和秋水仙碱组肝组织损伤减轻,胶原纤维沉积减少(P<0.05,P<0.01),CollagenⅠ、α-SMA、Beclin-1、LC3-Ⅱ蛋白表达水平显著降低(P<0.05,P<0.01)。结论疏肝健脾方具有抗肝纤维化作用,其机制与减少细胞自噬,降低能量的生成,从而抑制肝星状细胞活化有关。
Objective To observe the effect of Shugan Jianpi Recipe（ SGJPR） on autophagy protein Beclin-1 and LC3-Ⅱ in liver fibrosis rats. Methods Sixty Sprague Dawley（ SD） rats were randomly divided into six groups,the normal group,the model group,high,middle,and low dose SGJPF groups（ 1. 5,3. 0,6. 0 g/kg）,and the colchicines group（ 0. 1× 10^（-3）g/kg）,10 in each group. Except those in the normal group,liver fibrosis model was induced by subcutaneous injection of CCl4 to rats in the rest groups,twice per week for 12 consecutive weeks. At week 7 after modeling,different doses of SGJPF and colchicines were respectively administrated to rats in corresponding groups by gastrogavage,once per day for 6 weeks. Equal volume of saline solution was administrated to rats in the normal group and the model group,once per day for 6 weeks. The pathological injury degrees of liver were observed by HE staining and Masson dyeing. Expressions of Collagen Ⅰ,α-SMA,Beclin-1,and LC3-Ⅱ protein were detected by immunohistochemistry. Results Compared with the normal group,the histopathological injury degree of the liver was obviously aggravated,the expressions of Collagen Ⅰ,α-SMA,Beclin-1,and LC3-Ⅱ protein in liver tissue were significantly elevated in the model group（P〈0. 01）. Compared with the model group,liver injury was attenuated and collagen deposition was reduced（P〈0. 05,P〈0. 01）,expressions of Collagen Ⅰ,α-SMA,Beclin-1,and LC3-Ⅱ protein in liver tissue were significantly decreased（P〈0. 05,P〈0. 01） in middle and low dose SGJPF groups and the colchicines group. Conclusions SGJPF could fight against liver fibrosis. Its mechanism might be associated with reducing autophagy and lowering energy generation,thus inhibiting activation of hepatic stellate cells.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2017年第8期955-960,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81102874)
安徽省卫生计生委中医药科研资助项目(No.2014zy09)
安徽省高校领军人才引进与培育计划资助项目(No.gxfx ZD2016118)
关键词
疏肝健脾方
肝纤维化
肝星状细胞
自噬
Shugan Jianpi Recipe
liver fibrosis
hepatic stellate cell
autophagy
