基于调控外泌体释放的电针对慢传输型便秘大鼠结肠组织TRPM7-PI3K/Akt通路的影响

Effects of Electroacupuncture on TRPM7-PI3K/Akt Signaling Pathway in Colonic Tissue of Rats with Slow Transit Constipation Based on Modulating the Release of Exosomes

目的观察抑制血清外泌体释放对电针调节慢传输型便秘(STC)大鼠结肠组织TRPM7-PI3K/Akt信号通路的影响,探讨电针治疗STC的作用机制。方法50只SD大鼠随机分为对照组、模型组、电针组、药物组和电针+GW4869组,每组10只。除对照组外,采用盐酸洛哌丁胺混悬液灌胃7 d制备STC大鼠模型,成模后分别予电针或药物干预2周,电针+GW4869组在电针基础上隔日腹腔注射GW4869混悬液。采用超速离心法提取血清外泌体,电镜观察外泌体形态;检测大鼠24 h排便量;Western blot检测结肠组织瞬时受体电位M7(TRPM7)、磷酸肌醇-3-激酶(PI3K)、蛋白激酶B(Akt)、p-Akt蛋白表达;RT-PCR检测结肠组织TRPM7、PI3K、Akt mRNA表达。结果与对照组比较,模型组大鼠第14、21日24 h排便量明显减少(P<0.01),结肠组织TRPM7、PI3K、Akt蛋白和mRNA表达及p-Akt/Akt比值明显降低(P<0.01);与模型组比较,电针组和药物组大鼠第14、21日24 h排便量明显增加(P<0.01),结肠组织TRPM7、PI3K、Akt蛋白和mRNA表达及p-Akt/Akt比值明显升高(P<0.01);与电针组比较,电针+GW4869组大鼠第14、21日24 h排便量明显减少(P<0.01),结肠组织TRPM7、PI3K、Akt蛋白和mRNA表达及p-Akt/Akt比值明显降低(P<0.05,P<0.01)。结论电针可通过调节外泌体释放激活TRPM7-PI3K/Akt信号通路,改善STC大鼠排便情况。

Objective To observe the effect of blocking serum exosomes on regulating TRPM7-PI3K/Akt signaling pathway in colonic tissue of rats with slow transit constipation(STC)by electroacupuncture;To explore the mechanism of electroacupuncture in the treatment of STC.Methods Totally 50 SD rats were randomly divided into control group,model group,electroacupuncture group,drug group,and electroacupuncture+GW4869 group,with 10 rats in each group.Except the control group,STC rat model was established by intragastric administration of loperamide hydrochloride suspension for 7 days.After modeling,the intervention was given electroacupuncture or drug treatments for 2 weeks respectively.The electroacupuncture+GW4869 group was intraperitoneal injection of GW4869 to inhibit the release of exosomes.The exosomes were extracted from serum by supercentrifugation and the morphological characteristics were observed by electron microscopy,the weight of 24-hour defecation was detected,the expressions of TRPM7,PI3K,Akt,and p-Akt protein in colonic tissue were detected by Western blot,the expressions of TRPM7,PI3K,and Akt mRNA in colonic tissue were detected by RT-PCR.Results Compared with the control group,the weight of 24-hour defecation on the 14th and 21st days in model group reduced significantly(P<0.01),the expressions of TRPM7,PI3K,Akt protein and mRNA,and the ratio of p-Akt/Akt in colonic tissue in model group decreased significantly(P<0.01).Compared with the model group,the weight of 24-hour defecation on the 14th and 21st days in electroacupuncture group and drug group increased significantly(P<0.01),the expressions of TRPM7,PI3K,Akt protein and mRNA,and the ratio of p-Akt/Akt in colonic tissue increased significantly(P<0.01).Compared with the electroacupuncture group,the weight of 24-hour defecation on the 14th and 21st days in electroacupuncture+GW4869 group reduced significantly(P<0.01),the expressions of TRPM7,PI3K,Akt protein and mRNA,and the ratio of p-Akt/Akt in colonic tissue decreased(P<0.05,P<0.01).Conclusion Electroacupuncture can activate the TRPM7-PI3K/Akt signaling pathway by regulating the release of exosomes and improve the defecation of STC rats.