刺山柑果实提取物对CCl4诱导的小鼠肝纤维化的保护作用  

Protective effects of extract of Capparis spinosa L. fructus on CCl4-induced liver fibrosis in mice

作　　者：柴琼 柳雨婷 刘伟 瞿惠燕 周华 杨涛[2,3,4] CHAI Qiong;LIU Yuting;LIU Wei;QU Huiyan;ZHOU Hua;YANG Tao(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Cardiology,Institute of Cardiovascular Disease,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Liver Diseases,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海中医药大学附属曙光医院心内科、心血管病研究所,上海201203 [3]上海中医药大学附属曙光医院肝病研究所,上海201203 [4]上海市中医临床重点实验室,上海201203

出　　处：《上海中医药大学学报》2019年第5期48-53,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：上海市科委科研计划资助项目(18401900700);上海市申康医院发展中心新兴前沿技术联合攻关项目(SHDC120180125)

摘　　要：目的:研究刺山柑果实提取物对四氯化碳(CCl4)诱导的小鼠肝纤维化的保护作用及可能机制。方法:45只C57BL小鼠随机分为正常组、模型组、扶正化瘀方组(FZHY组,4 g/kg)、刺山柑低剂量组(0.93 g/kg)和高剂量组(2.80 g/kg),每组9只。除正常组外,其余各组小鼠腹腔注射10%CCl4,每次2 ml/kg,每周3次,连续8周,诱导小鼠实验性肝纤维化模型。造模第5周起,各药物组灌胃给予相应剂量的药液,连续4周。末次给药后,采集血清和肝脏样本。试剂盒检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)活性以及总胆红素(TBIL)、白蛋白(ALB)、总蛋白(TP)水平。HE染色和天狼猩红染色后,光镜下观察肝组织形态学变化。试剂盒检测肝组织羟脯氨酸(Hyp)水平以及超氧化物歧化酶(SOD)、丙二醛(MDA)活性。Western blot检测肝组织自噬相关蛋白LC3-Ⅰ和LC3-Ⅱ的表达水平。结果:①与模型组相比,FZHY组和刺山柑低、高剂量组小鼠的ALT、AST、ALP、TBIL水平显著降低(P<0.05,P<0.01),TP和ALB水平显著升高(P<0.01);且刺山柑高剂量组小鼠的ALT和AST水平明显低于低剂量组(P<0.05,P<0.01)。②HE染色和天狼猩红染色观察显示,模型组小鼠肝细胞可见大面积出血性坏死,纤维组织广泛增生形成间隔,并将肝组织包围形成假小叶;与模型组相比,FZHY组及刺山柑低、高剂量组小鼠的肝组织形态均有不同程度改善,肝细胞坏死明显减少,同时纤维增生有所减轻,假小叶逐渐消失;且刺山柑高剂量组的改善作用优于低剂量组。半定量分析结果表明,FZHY组和刺山柑高剂量组的胶原阳性染色面积占比较模型组显著降低(P<0.05)。③与模型组相比,FZHY组及刺山柑低、高剂量组小鼠肝组织Hyp含量显著降低(P<0.01),SOD水平显著升高(P<0.01),MDA水平显著降低(P<0.05,P<0.01)。④Western blot结果显示,刺山柑低、高剂量组小鼠肝组织LC3-Ⅱ蛋白表达较Objective: To study the protective effects and possible mechanism of extract of Capparis spinosa L. fructus on carbon tetrachloride(CCl4)-induced liver fibrosis in mice. Methods: Forty-five C57 BL mice were randomly divided into the normal group, model group, Fuzheng Huayu(FZHY, 4 g/kg) group, Capparis spinosa L. groups with low-(0.93 g/kg) and high-(2.80 g/kg) dose, 9 mice in each group. Except the normal group, the other groups were treated with 10% CCl4 by intraperitoneal injection at dose of 2 ml/kg, 3 times a week for 8 weeks, in order to induce the liver fibrosis model in mice. From the 5 th week of modeling, the drug treatment groups were treated with the corresponding drugs by intragastric administration for 4 weeks. After the last administration, the serum and liver samples were collected. The serum levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP), total bilirubin(TBIL), albumin(ALB) and total protein(TP) were detected by kit. After HE staining and Sirius red staining, the morphological changes of liver tissue were observed under light microscopy. The levels of hydroxyproline(Hyp) and the activities of superoxide dismutase(SOD) and malondialdehyde(MDA) in liver tissue were detected by kit. The expressions of autophagy related proteins LC3-Ⅰ and LC3-Ⅱ in liver tissue were detected by Western blot. Results: ①Compared with the model group, the serum levels of ALT,AST, ALP and TBIL were significantly decreased in the FZHY group and Capparis spinosa L. groups with low-and high-dose(P<0.05, P<0.01), and the levels of TP and ALB were significantly increased(P<0.01). The levels of ALT and AST in the Capparis spinosa L. group with high-dose were significantly lower than those in the low-dose group(P<0.05, P<0.01). ②HE staining and Sirius red staining showed that massive hemorrhagic necrosis of hepatocytes was observed in the model group, as well as the extensive proliferation of fibrous tissue with spaces, and the proliferated fibers surrounded the liver

关 键 词：刺山柑 肝纤维化 CCL4 自噬 小鼠 

分 类 号：R285.5[医药卫生—中药学]