川芎嗪联合丹皮酚对CCl_4致大鼠肝纤维化的抑制作用及其机制研究  

The inhibitory effect of ligustrazine combined with paeonol on rat liver fibrosis induced by CCl_4 and mechanisms

作　　者：孔德松[1] 张自力[2,3] 张峰[2,3] 邱萍[4] 郑仕中[2,3] 

机构地区：[1]南京中医药大学第三附属医院,江苏南京210001 [2]南京中医药大学药学院,江苏南京210023 [3]江苏省中药药效与安全性评价重点实验室,江苏南京210023 [4]浙江中医药大学药学院,浙江杭州310053

出　　处：《中国药理学通报》2016年第12期1741-1746,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81270514;31401210;31571455;81600483);江苏高校优势学科建设工程资助项目(No ysxk-2010);江苏省自然科学基金青年基金资助项目(No BK20140955);江苏省高校自然科学研究面上项目(No 14KJB310011);南京市医学科技发展基金项目(No YKK14143)

摘　　要：目的探索川芎嗪联合丹皮酚对CCl_4致大鼠肝纤维化的抑制作用及其机制,为临床肝纤维化提供新的治疗策略。方法清洁级♂SD大鼠,随机分为空白对照组、模型组(CCl_4)、川芎嗪组、丹皮酚组以及联合用药组(川芎嗪+丹皮酚)。HE染色观察病理组织学改变,Masson染色及天狼星红染色观察胶原沉积情况,全自动生化仪检测血清中ALT、AST、ALP和羟脯氨酸,Western blot检测肝纤维化标志物。HSC-T6细胞分为模型组、川芎嗪组、丹皮酚组以及联合用药组,Western blot与Real time-PCR检测炎症及凋亡蛋白与基因的表达情况。结果川芎嗪与丹皮酚联合应用能明显改善CCl_4所致肝组织病理改变,降低血清ALT、AST、ALP及羟脯氨酸水平,抑制肝纤维化标志蛋白表达,进而减少胶原纤维的沉积,效果明显优于单独干预组。联合用药组能抑制HSC中炎症通路相关蛋白表达,促进HSC的凋亡。结论川芎嗪与丹皮酚联合应用具有明显的抗肝纤维化效应,且效果优于两者单独干预,这种效果可能是由于干扰了HSC中TNF-α/NF-κB通路,进而促进细胞凋亡,抑制了细胞外基质的生成。Aim To explore the inhibitory effect of li- gustrazine combined with paeonol on rat liver fibrosisinduced by CCl4 and its mechanisms,so as to provide new treatment strategies for liver fibrosis in clinical.Methods Cleaning laboratory male SD rats were randomly divided into blank control group,model group（ CCl4）,ligustrazine group,paeonol group and combination group（ ligustrazine ＋ paeonol）. HE staining was used to observe the pathological change. Masson staining and Sirius red staining was used to observe the collagen deposition. The levels of serum ALT,AST,ALP and hydroxyproline were detected by automatic biochemistry analyzer. Western blot detected the markers of liver fibrosis. HSC-T6 cell was divided into model group,ligustrazine group,paeonol group and combination group. The protein and gene expression of inflammation and apoptosis pathway was analyzed by Western blot and real time-PCR. Results Ligustrazine combined with paeonol could significantly improve liver tis-sue pathology changes caused by CCl4. It could reduce serum ALT, AST, ALP and hydroxyproline levels.Moreover,it could also inhibit liver fibrosis marker protein expression,and thus reduce the deposition of collagen fibers. The effect was better than that in single intervention group. Combination group could inhibit the inflammatory pathways related protein expression in HSC cells and promote the apoptosis of HSC cells.Conclusion Ligustrazine in combination with paeonol has significant anti-fibrosis effect,and the effect is better than both single intervention. The effect may be due to the interference with TNF-α / NF-κB pathway in the HSC cells,which promotes its apoptosis and inhibits the generation of extracellular matrix.

关 键 词：川芎嗪 丹皮酚 肝纤维化 炎症 凋亡 细胞外基质 

分 类 号：R-332[医药卫生] R284.1