桃红四物汤对大鼠肢体缺血-再灌注损伤骨骼肌Wnt5a信号表达的影响  

Effect of Taohong Siwu Decoction(桃红四物汤) on Wnt5a Signal Expression in Skeletal Muscle of Rats with Limb Ischemia-Reperfusion Injury

作　　者：李武平[1] 李婕[2] 易南 黄思琦 刘宗义 朱付平[1] LI Wu-ping;LI Jie;YI Nan;HUANG Si-qi;LIU Zong-yi;ZHU Fu-ping(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China;Hunan University of Chinese Medicine,Changsha Hunan 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,长沙湖南410007 [2]湖南中医药大学,长沙湖南410208

出　　处：《中医药导报》2021年第5期23-28,39,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(81674008);湖南省自然科学基金项目(2020JJ4070);湖南省教育厅科学研究基金项目(20C1426)。

摘　　要：目的:观察桃红四物汤对肢体缺血-再灌注损伤大鼠骨骼肌Wnt5a信号表达的影响,从Wnt/Ca^(2+)信号通路角度探讨其作用机制。方法:100只8周龄雄性SD大鼠随机分为正常组、模型组、桃红四物汤低剂量组、桃红四物汤高剂量组、阻滞剂组,每组20只。除正常组外,其他各组均阻断血流复制右后肢缺血-再灌注损伤模型,各药物组大鼠在造模前分别以相应药物预处理,ELISA检测复灌后0、2、4、8 h大鼠腓肠肌中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的水平,RT-qPCR检测复灌后0、2、4、8 h腓肠肌Wnt5a mRNA的表达,免疫组化检测复灌后8 h腓肠肌中Wnt5a蛋白的阳性表达。结果:HE染色显示缺血-再灌注8 h后,桃红四物汤低剂量组、桃红四物汤高剂量组及阻滞剂组病理形态学优于模型组。不同再灌注时间大鼠腓肠肌IL-1β表达水平比较,差异无统计学意义(P>0.05),不存在时间效应;不同再灌注时间大鼠腓肠肌TNF-α、Wnt5a mRNA表达水平比较,差异均有统计学意义(P<0.05),存在时间效应。5组大鼠腓肠肌IL-1β、TNF-α及Wnt5a mRNA表达水平总体比较,差异均有统计学意义(P<0.05),存在分组效应。时间与组别对大鼠腓肠肌IL-1β、TNF-α及Wnt5a mRNA表达水平的影响存在交互作用(P<0.05)。与正常组比较,模型组大鼠腓肠肌IL-1β、TNF-α表达升高(P<0.05),Wnt5a蛋白和Wnt5a mRNA表达一致性上调(P<0.05);与模型组比较,阻滞剂组、桃红四物汤低剂量组和桃红四物汤高剂量组大鼠腓肠肌IL-1β、TNF-α表达降低(P<0.05),Wnt5a蛋白和Wnt5a mRNA表达一致性下调(P<0.05),不同再灌注时间(0、2、4、8 h)的结果显示相同的倾向性。结论:Wnt5a信号为大鼠肢体缺血-再灌注骨骼肌损伤的重要信号分子,桃红四物汤干预可在一定程度上下调Wnt5a信号的表达来减轻肢体缺血-再灌注损伤。Objective:To observe the effect of Taohong Siwu decoction on Wnt5 a signal expression in skeletal muscle of rats with limb ischemia-reperfusion injury,and to explore its mechanism from the perspective of Wnt/Ca^(2+) signaling pathway.Methods:A total of 1008-week-old male SD rats were randomly divided into normal group,model group,Taohong Siwu decoction low-dose group,Taohong Siwu decoction high-dose group,and blocker group,with 20 rats in each group.Except for the normal group,the right hind limb ischemia-reperfusion injury model was reproduced by blocking blood flow in other groups.The rats in each drug group were pretreated with corresponding drugs before modeling.The levels of IL-1β and TNF-α in gastrocnemius muscle were detected by ELISA at 0,2,4 and 8 h after reperfusion.RT qPCR was used to detect the expression of Wnt5 a mRNA in gastrocnemius at 0,2,4 and 8 h after reperfusion,and immunohistochemistry was used to detect the positive expression of Wnt5 a protein in gastrocnemius at 8 h after reperfusion.Results:HE staining showed that after 8 hours of ischemia-reperfusion,the pathomorphology of Taohong Siwu decoction low-dose,Taohong Siwu decoction high-dose and blocker groups was better than that of model group.There was no significant difference in the expression of IL-1β in gastrocnemius muscle at different time points after reperfusion(P>0.05),there was no time effect.There were significant differences in the expression of TNF-αand Wnt5 a mRNA in gastrocnemius muscle at different time points after reperfusion(P<0.05),there was time effect.There were significant differences in the expression of IL-1β,TNF-α and Wnt5 a mRNA among the five groups(P<0.05),there was group effect.The interaction of time and group had significant effect on the expression of IL-1β,TNF-α and Wnt5 a mRNA in gastrocnemius(P<0.05).Compared with the normal group,the levels of IL-1β and TNF-α in gastrocnemius muscle of model group were increased(P<0.05),and the expression of Wnt5 a protein and mRNA were up-regulated(P<0.05).C

关 键 词：肢体缺血再灌注损伤 桃红四物汤 炎症因子 Wnt5a信号 大鼠 

分 类 号：R285.5[医药卫生—中药学]