中药成分复方JY5通过抑制胆管反应抗胆汁性肝纤维化的研究  

Study of the Chinese Herbal Compound JY5 Against Biliary Hepatic Fibrosis through Inhibition of Ductular Reaction

作　　者：付亚东 胡永红 肖准 陈龙 刘伟[1] 慕永平[1] 陈佳美[1] 刘平[1,2] FU Yadong;HU Yonghong;XIAO Zhun;CHEN Long;LIU Wei;MU Yongping;CHEN Jiamei;LIU Ping(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Institute of Liver Diseases,Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China;Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Mediceine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海市中医药研究院肝病研究所,肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203 [2]上海中医药大学交叉科学研究院,上海201203

出　　处：《中药新药与临床药理》2022年第10期1298-1306,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金重点项目(81530101);上海市青年科技启明星计划项目(19QA1408900);中国博士后科学基金面上资助项目(2021M702218)。

摘　　要：目的探讨源自扶正化瘀方的中药成分复方JY5(丹酚酸B、苦杏仁苷和五味子酯甲)对胆管结扎诱导的大鼠肝纤维化的作用及机制。方法将32只SD大鼠随机分为假手术组、模型组、JY5组和DAPT组,每组8只。采用胆管结扎术制备大鼠胆汁性肝纤维化模型。胆管结扎术后1周开始给药,JY5组给予JY5混悬液(丹酚酸B 16 mg·kg^(-1)+苦杏仁苷0.5 mg·kg^(-1)+五味子酯甲2 mg·kg^(-1))10 mL·kg^(-1)灌胃给药;DAPT组按照30 mg·kg^(-1)腹腔注射给药;假手术组、模型组灌胃给予0.3%CMC-Na;每日1次,连续给药3周。检测大鼠血清肝功能相关生化指标;采用苏木素-伊红(HE)及天狼星红(SR)染色法观察肝组织炎性损伤和胶原沉积情况;碱水解法测定肝组织羟脯氨酸(Hyp)含量;qPCR、Western Blot法检测肝组织CK19、CK7、EpCAM mRNA及蛋白表达;免疫组织化学法检测肝组织CK19、CK7、EpCAM、OV6蛋白表达情况;免疫荧光染色法观察肝组织中CK19、OV6共定位情况。结果与假手术组比较,模型组大鼠血清中的丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(GGT)活性及总胆汁酸(TBA)、总胆红素(TBIL)、直接胆红素(DBIL)含量明显升高(P<0.01),白蛋白(ALB)含量明显降低(P<0.01);肝组织的炎性损伤和胶原沉积明显加重,肝脏SR染色阳性面积比和Hyp含量均明显升高(P<0.01);大鼠肝组织中CK19、CK7、EpCAM mRNA及蛋白表达量显著升高(P<0.01),OV6蛋白表达量显著升高(P<0.01);肝组织中OV6和CK19共表达的阳性细胞数显著增加。与模型组比较,JY5组大鼠血清的ALT、AST、ALP、GGT活性及TBA、TBIL、DBIL含量明显降低(P<0.01),ALB含量明显升高(P<0.01);肝组织的炎性损伤和胶原沉积明显减轻,肝脏SR染色阳性面积比和Hyp含量均明显降低(P<0.01);大鼠肝组织中CK19、CK7、EpCAM mRNA及蛋白表达量显著降低(P<0.01),OV6蛋白表达量显著降低(P<0.01);肝组织中OV6和CK1Objective To investigate the effect and mechanism of the Chinese herbal compound JY5(salvianolic acid B,amygdalin and Schisantherin A)on bile duct ligation(BDL)-induced liver fibrosis in rats. Methods Thirty-two SD rats were randomly divided into sham-operation group,model group,JY5 group and DAPT group,with 8 rats in each group. BDL was used to prepare the rat model of biliary hepatic fibrosis. The JY5 group was administrated1 week after BDL with JY5 suspension(salvianolic acid B 16 mg·kg^(-1)+ amygdalin 0.5 mg·kg^(-1)+ Schisantherin A2 mg·kg^(-1))at 10 mL·kg^(-1)by gavage;DAPT group was administrated at 30 mg·kg^(-1)by intraperitoneal injection;the sham-operation group and model group were given 0.3% CMC-Na by gavage;the treatments in all the groups were administrated once per day for consecutive 3 weeks. The rats were tested for biochemical parameters related to serum liver function;the inflammatory damage and collagen deposition in liver tissue were observed by hematoxylin-eosin(HE) and Sirius red(SR) staining;the hydroxyproline(Hyp) content of liver tissue was detected by alkaline hydrolysis;the m RNA and protein expressions of CK19,CK7 and EpCAM in liver tissue were detected by qPCR and Western Blot;the protein expressions of CK19, CK7, EpCAM and OV6 in liver tissue were detected by immunohistochemistry;the co-localization of CK19 and OV6 in liver tissue was observed by immunofluorescence staining. Results Compared with the sham-operation group,the serum levels of ALT,AST,ALP,GGT and the levels of TBA, TBIL and DBIL were significantly increased(P<0.01) and the level of ALB was significantly decreased(P<0.01) in the model group;the inflammatory damage and collagen deposition in liver tissue were significantly increased, and the ratio of positive area of SR staining and Hyp content in liver were significantly increased(P<0.01);the mRNA and protein expressions of CK19, CK7, EpCAM in rat liver tissues were significantly increased(P<0.01)and OV6 protein expression was significantly increased(P<0.01);the num

关 键 词：中药成分复方JY5 丹酚酸B 苦杏仁苷 五味子酯甲 胆管结扎 肝祖细胞 胆管反应 胆汁性肝纤维化 大鼠 

分 类 号：R285.5[医药卫生—中药学]