摘要
目的探究竹沥对肝纤维化小鼠Ras同源基因-Rho相关螺旋卷曲蛋白激酶(Ras homologousgene-Rho-associatedcoiled-coilcontainingkinases,Rho-ROCK)信号转导通路的作用,分析竹沥抗肝纤维化的分子机制。方法将40只KM小鼠随机分为空白组、模型组、竹沥组和丹参酚酸B组,每组10只,模型组、竹沥组和丹参酚酸B组小鼠给予40%CCl4花生油混合液(剂量2μl/g),空白组给予等量生理盐水,腹腔注射,2次/周,连续8周。模型构建成功后,竹沥组以竹沥灌胃,每日1次,每次30 ml/100 g;丹参酚酸B组以丹参酚酸B溶液灌胃,每日1次,每次用量0.25 mg/100 g;其余2组给予等量生理盐水灌胃,持续4周。末次给药后对小鼠眼球取血并处死,采用HE染色观察肝组织病理并进行Ishak评分,采用γ放射免疫全自动双探头计数器检测肝纤维化指标,包括透明质酸、层粘连蛋白、Ⅲ型前胶原和Ⅳ型胶原,采用实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction,q RT-PCR)检测Rho A和ROCK2m RNA的相对表达量,采用Western blot检测Rho A和ROCK2蛋白的相对表达量。结果空白组、模型组、竹沥组和丹参酚酸B组小鼠Ishak评分分别为(0.40±0.05)分、(4.80±0.84)分、(3.20±0.45)分和(2.80±0.83)分,差异有统计学意义(F=34.807,P<0.001),其中空白组显著低于模型组、竹沥组和丹参酚酸B组(t=8.690,P<0.001;t=5.674,P<0.001;t=4.768,P=0.001),模型组显著高于竹沥组和丹参酚酸B组(t=3.016,P=0.017;t=3.922,P=0.004),竹沥组和丹参酚酸B组差异无统计学意义(t=0.809,P=0.464)。空白组小鼠肝纤维化指标均显著低于模型组、竹沥组和丹参酚酸B组[透明质酸:(26.14±2.69)μg/L vs(53.69±5.87)μg/L vs(37.21±3.04)μg/L vs(32.88±3.60)μg/L,层粘连蛋白:(120.37±15.57)μg/L vs(214.48±21.39)μg/L vs(162.54±19.22)μg/L vs(161.66±14.23)μg/L,Ⅲ型前胶原:(4.04±1.01)μg/L vs(11.70±3.09)μg/L vs(8.12±1.87)μg/L vs(7.80±1.72)μg/L,Ⅳ型胶原:(12.96±2.82)μg/L vs
Objective To investigate the effects of Zhuli on the Ras homologous gene-Rhoassociated coiled-coil containing kinases(Rho-ROCK)signaling transduction pathway in mice with liver fibrosis and to analyze the molecular mechanism of Zhuli against liver fibrosis.Methods A total of forty KM mice were divided into blank group,model group,Zhuli group and salvianolic acid B group,10 mice in each group.Mice in model group,Zhuli group and salvianolic acid B group were given 40%CCl4 peanut oil mixture(2μl/g)and mice in blank group were given equal amounts of saline by intraperitoneal injection for 8 weeks.After the successful model construction,mice in Zhuli group were gavaged with Zhuli once a day(30 ml/100 g),mice in salvianolic acid B group were gavaged with salvianolic acid B once a day(0.25 mg/100 g)and mice in the other two groups were gavaged with equal amounts of saline for 4 weeks.After the last administration,blood was taken from the eyeballs and the mice were sacrificed.Liver histopathology was visualized by HE staining and Ishak score was calculated.Liver fibrosis indexes were detected by a fully automated double probe counter,including hyaluronic acid,laminin,procollagenⅢandⅣcollagen.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the m RNA relative expression of Rho A and ROCK2.Western blot was used to detect the protein relative expression of Rho A and ROCK2.Results The Ishak score of mice in blank group,model group,Zhuli group and salvianolic acid B group were(0.40±0.05)points,(4.80±0.84)points,(3.20±0.45)points and(2.80±0.83)points,respectively,the difference was statistically significant(F=34.807,P<0.001),the Ishak score of mice in blank group was significantly lower than that in model group,Zhuli group and salvianolic acid B group(t=8.690,P<0.001;t=5.674,P<0.001;t=4.768,P=0.001),the Ishak score of mice in model group was significantly higher than that in Zhuli group and salvianolic acid B group(t=3.016,P=0.017;t=3.922,P=0.004),and there was no significant difference of I
作者
王玖恒
徐成军
王晓宏
张小冰
刘青松
王云中
郑洋
赵铁建
段雪琳
Wang Jiuheng;Xu Chengjun;Wang Xiaohong;Zhang Xiaobing;Liu Qingsong;Wang Yunzhong;ZHENG Yang;Zhao Tiejian;Duan Xuelin(Department of Gastroenterology,Kuancheng Manchu Autonomous County Hospital,Chengde 067600,Hebei Province,China;Faculty of Chinese Medicine Science,Guangxi University of Chinese Medicine,Nanning 530222,China;Department of Physiology,College of Basic Medicine,Guangxi University of Chinese Medicine,Nanning 530222,China;Zhuang Medical College,Guangxi University of Chinese Medicine,Nanning 530222,China)
出处
《中国肝脏病杂志（电子版）》
CAS
2021年第4期38-45,共8页
Chinese Journal of Liver Diseases：Electronic Version
基金
国家自然科学基金项目(81960751)
广西中青年教师科研基础能力提升项目(2020KY59009)
广西壮瑶药重点实验室项目(GXZYZZ2020-07)
广西中医药大学一流学科项目(2019XK141)。
