摘要
目的:从血管新生角度探讨补肾柔肝方抗小鼠肝纤维化作用机制。方法:雄性C57小鼠按体质量分层随机分为正常组、模型组、补肾柔肝方组、索拉非尼(Sorafenib)组。模型组、补肾柔肝方组、索拉非尼组以0.4%二甲基亚硝胺(DMN)腹腔注射,1次/d,每周3次,共8周,诱导肝纤维化模型。造模第5周开始给药,8周后处死留取血清、肝组织样本。记录小鼠体质量变化及死亡情况;取材后检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及白蛋白(ALB)含量。以天狼猩红胶原染色和肝组织羟脯氨酸(Hyp)含量评估肝纤维化程度;以肝组织血小板内皮细胞粘附分子-1(CD31)和血管内皮生长因子受体2(VEGFR2)蛋白表达情况评价肝脏血管新生情况。结果:模型组小鼠死亡率为15.4%(2/13),补肾柔肝方组无小鼠死亡,索拉非尼组小鼠死亡率7.7%(1/13)。补肾柔肝方组及索拉非尼组小鼠血清ALT、AST含量较模型组降低(P<0.05),血清ALB含量升高(P<0.05);补肾柔肝方组和索拉非尼组小鼠肝组织Hyp含量较模型组显著降低(P<0.05)。天狼星红染色显示,补肾柔肝方组和索拉非尼组纤维化程度较模型组明显减轻;Western blot及肝组织免疫组化实验显示,补肾柔肝方组和索拉菲尼组小鼠肝组织CD31和VEGFR2蛋白表达较模型组均显著下调(P<0.05)。结论:补肾柔肝方可有效改善DMN诱导小鼠模型肝纤维化,其作用机制可能与抑制血管新生有关。
Objective:To investigate the mechanism of Bushen Rougan Formula against hepatic fibrosis from the perspective of angiogenesis.Methods:Male C57 mice were stratified randomly divided into normal group,model group,Bushen Rougan Formula group,Sorafenib group according to body weight.In order to induce liver fibrosis model,the model group,Bushen Rougan Formula group and Sorafenib group were intraperitoneally injected with 0.4%dimethylnitrosamine(DMN)once a day,three times a week for 8 weeks.The drug was administered at the 5th week of modeling.After 8 weeks,the mice were sacrificed,and serum and liver tissue samples were collected.The changes of body weight and death of mice were recorded.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and albumin(ALB)contents were detected after sampling.The degree of hepatic fibrosis was evaluated by sirius red collagen staining and liver hydroxyproline(Hyp)contents.The expression of platelet endothelial cell adhesion molecule-1(CD31)and vascular endothelial growth factor receptor 2(VEGFR2)protein in liver tissue were used to evaluate hepatic angiogenesis.Results:The mortality rate of mice in the model group,Bushen Rougan Formula group and the Sorafenib group was 15.4%(2/13),0 and 7.7%(1/13),respectively.The contents of serum ALT and AST in Bushen Rougan Formula group and the Sorafenib group were significantly decreased(P<0.05),while the content of serum ALB was significantly increased(P<0.05).Sirius red staining showed that the degree of fibrosis in Bushen Rougan Formula group and Sorafenib group was significantly reduced compared with the model group.Western blot and liver immunohistochemistry showed that CD31 and VEGFR2 protein expression were significantly down-regulated in Bushen Rougan Formula group and the Sorafenib group compared with the model group(P<0.05).Conclusion:Bushen Rougan Formula can effectively improve DMN-induced hepatic fibrosis in mouse model,and its mechanism may be related to inhibition of angiogenesis.
作者
王欣
安梓铭
董慧琳
聂红明
WANG Xin;AN Ziming;DONG Huilin;NIE Hongming(Institute of Hepatology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Hepatology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《上海中医药大学学报》
CAS
2022年第S01期167-173,共7页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81373618)
上海中医药大学附属曙光医院四明青年基金项目(SGKJ-201703)
关键词
补肾柔肝方
二甲基亚硝胺
肝纤维化
血管新生
Bushen Rougan Formula
dimethylnitrosamine
hepatic fibrosis
angiogenesis
