金银花-连翘提取物组合对CCl4诱导小鼠肝纤维化的改善作用研究  

Protective effects of extract combination of Lonicerae Japonicae Flos and Forsythiae Fructus against CCl-induced liver fibrosis in mice

作　　者：阎仕豪 苗蕙 黄镇林 季莉莉[1] YAN Shihao;MIAO Hui;HUANG Zhenlin;JI Lili(MOE Key Laboratory for Standardization of Chinese Medicine,Shanghai Key Laboratory of Compound Chinese Medicines,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学,中药研究所中药标准化教育部重点实验室,上海市复方中药重点实验室,上海201203

出　　处：《上海中医药大学学报》2021年第2期50-56,共7页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家重点研发计划“中医药现代化研究”重点专项项目(2018YFC1707302)。

摘　　要：目的:探讨金银花-连翘提取物组合对四氯化碳(CCl4)诱导的小鼠肝纤维化的改善作用。方法:分别制备金银花和连翘提取物,混合后制得组合物(药材配比1∶2)。取雄性C57小鼠随机分为对照组、CCl4组、组合低剂量(0.5 g/kg)+CCl4组、组合高剂量(1.0 g/kg)+CCl4组、组合高剂量(1.0 g/kg)组,每组5只。除对照组和组合高剂量组外,其他各组小鼠腹腔注射CCl4诱导肝纤维化,每周2次,共4周。同时,各组灌胃给予相应剂量的组合物,连续4周。末次给药后,取血,收集肝脏组织。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)水平以及肝脏羟脯氨酸(Hyp)、Ⅳ型胶原(ColⅣ)含量。HE染色观察肝组织形态学变化,天狼猩红染色和Masson染色观察肝脏胶原沉积。PCR检测肝组织胶原1a1(Col1a1)、胶原3a1(Col3a1)、纤维连接蛋白(FN)、转化生长因子β(TGF-β)及α-平滑肌肌动蛋白(α-SMA)的mRNA表达,Western blot检测肝组织α-SMA蛋白表达。结果:(1)与CCl4组相比,组合高剂量+CCl4组血清ALT、AST水平显著降低(P<0.01),肝组织炎症反应和肝细胞坏死程度明显减轻。(2)与CCl4组相比,组合高剂量+CCl4组肝脏Hyp和ColⅣ含量显著降低(P<0.05,P<0.01),组合低剂量+CCl4组ColⅣ含量亦显著降低(P<0.05),且两组肝组织胶原沉积程度明显减轻。(3)不同剂量的金银花-连翘组合物干预后,可明显下调模型小鼠肝脏Col1a1、Col3a1、FN和TGF-β的mRNA表达(P<0.05,P<0.01),α-SMA的mRNA和蛋白表达亦明显降低(P<0.05,P<0.01)。(4)金银花-连翘组合物对正常小鼠肝组织形态及以上指标无明显影响。结论:金银花-连翘提取物组合可能通过抑制肝星状细胞活化,减少胶原沉积,从而改善肝纤维化。Objective: To investigate the effects of extract combination of Lonicerae Japonicae Flos and Forsythiae Fructus on liver fibrosis induced by carbon tetrachloride(CCl4) in mice. Methods: The extracts of Lonicerae Japonicae Flos and Forsythiae Fructus were prepared respectively. After mixing, the combination was obtained(the ratio of medicinal materials as 1∶2). Male C57 mice were randomly divided into the control group, CCl4 group, combination low-dose(0.5 g/kg) + CCl4 group, combination high-dose(1.0 g/kg) + CCl4 group and combination high-dose(1.0 g/kg) group, 5 mice in each group. Except the control group and combination high-dose group, the mice in the other groups were intraperitoneally injected with CCl4 to induce liver fibrosis, twice a week for 4 weeks. Meanwhile,each group was treated with the corresponding dose of combination by intragastric administration for 4 weeks. After the last administration,the blood was taken and the liver tissue was collected. The levels of alanine aminotransferase( ALT) and aspartate aminotransferase( AST) in serum and the contents of hydroxyproline( Hyp) and collagen Ⅳ( Col Ⅳ) in liver were detected by kits. The morphological changes of liver tissue were observed after HE staining,and the deposition of liver collagen was observed after Sirius red staining and Masson staining. The mR NA expressions of collagen 1 a1( Col1 a1),collagen 3 a1( Col3 a1),fibronectin( FN),transforming growth factor-β( TGF-β) and α-smooth muscle actin( α-SMA) were detected by PCR,and the protein expression of α-SMA was detected by Western blot. Results:(1)Compared with the CCl4 group,the levels of ALT and AST in the combination high-dose + CCl4 group were significantly decreased( P<0.01),and the degree of liver inflammation and hepatocyte necrosis was significantly reduced.(2)Compared with the CCl4 group,the contents of Hyp and Col Ⅳ in liver tissue were significantly decreased in the combination high-dose + CCl4 group( P<0.05,P<0.01),and the content of Col Ⅳ was also significantly redu

关 键 词：金银花-连翘 组合 肝纤维化 胶原沉积 肝星状细胞 小鼠 

分 类 号：R285.5[医药卫生—中药学]