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三磷酸肌醇受体(IP3R)表达的影响,从Wnt/Ca2+信号通路角度探讨其作用机制。方法取2月龄雄性SD大鼠80只,随机分为正常组、模型组、桃红四物汤组和阻滞剂组,每组20只,除正常组外,其余各组均阻断血流制备大鼠右后肢缺血-再灌注损伤模型,桃红四物汤组、阻滞剂组造模前分别给予桃红四物汤灌胃、阻滞剂腹腔注射预处理,HE染色和DAPI荧光染色观察再灌后8 h腓肠肌细胞凋亡,免疫组化检测再灌后8 h腓肠肌IP3R蛋白的表达,RT-PCR检测再灌后0、2、4、8 h腓肠肌IP3R mRNA的表达。结果HE染色和DAPI荧光染色结果显示,与正常组比较,模型组大鼠腓肠肌细胞凋亡率明显升高(P<0.05);与模型组比较,桃红四物汤组和阻滞剂组大鼠腓肠肌细胞凋亡率明显降低(P<0.05),桃红四物汤组低于阻滞剂组(P<0.05)。免疫组化及RT-PCR检测结果显示,与正常组比较,模型组大鼠腓肠肌IP3R蛋白和m RNA表达明显上调(P<0.05);与模型组比较,桃红四物汤组、阻滞剂组大鼠腓肠肌IP3R蛋白和m RNA表达明显下调(P<0.05)。结论桃红四物汤通过下调Wnt5a/Ca2+信号通路IP3R的表达减轻肢体缺血-再灌注损伤。
Objective To observe the effects of Taohong Siwu Decoction on the expression of IP3R in skeletal muscle of rats after limb ischemia-reperfusion injury;To explore the mechanism of limb ischemia-reperfusion injury from the perspective of Wnt/Ca2+signaling pathway and the intervention effect of Taohong Siwu Decoction.Methods Totally 802-month-old male SD rats were randomly divided into normal group,model group,Taohong Siwu Decoction group and blocking agent group,with 20 rats in each group.Except for the normal group,the blood flow of other groups was blocked to establish the right hindlimb ischemia-reperfusion injury model.The Taohong Siwu Decoction group and the blocker group were given Taohong Siwu Decoction by gavage and intraperitoneal injection of blocker respectively.HE staining and DAPI staining were used to observe the apoptosis of gastrocnemius muscle cells at 8 h after reperfusion.Immunohistochemistry was used to observe the expression of IP3R in gastrocnemius muscle.RT-PCR was used to detect the expression of IP3R mRNA in gastrocnemius muscle at 0,2,4,8 h after reperfusion.Results HE staining and DAPI fluorescence staining results showed that,compared with the normal group,the apoptosis rate of model group was higher than that of normal group(P<0.05),the apoptosis rate of Taohong Siwu Decoction group and blocking agent group was lower than that of model group(P<0.05),and that of Taohong Siwu Decoction group was lower than that of blocker group(P<0.05).Results of immunohistochemistry and RT-PCR showed that,compared with the normal group,the expression of IP3R protein and mRNA in the model group were all significantly up-regulated(P<0.05).Compared with the model group,the expression of IP3R protein and m RNA in the Taohong Siwu Decoction group and the blocking agent group was significantly down-regulated(P<0.05).Conclusion Taohong Siwu Decoction can reduce the expression of IP3R by regulating Wnt5a/Ca2+signal pathway to alleviate limb ischemia-reperfusion injury.
作者
李武平
李婕
刘宗义
易南
黄思琦
朱付平
LI Wuping;LI Jie;LIU Zongyi;YI Nan;HUANG Siqi;ZHU Fuping(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中医药信息杂志》
CAS
CSCD
2021年第5期60-64,共5页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金(81674008)
湖南省自然科学基金(2020JJ4070)
湖南省教育厅科学研究基金项目(20C1426)。
作者简介
通讯作者:朱付平,E-mail:zhufp120@163.com。
