目的 观察半夏泻心汤血浆源性外泌体对胃电节律失常模型大鼠胃窦组织Cajal间质细胞(ICCs)网络结构的影响,探索其调节胃运动的可能作用机制.方法 共52只大鼠,选择20只完全随机分为中药组及阴性对照组,采用"隔日进食+饮稀盐酸"法建立胃电节律失常大鼠模型,半夏泻心汤及蒸馏水干预2周,分离血浆外泌体并鉴定;其余大鼠随机分为对照组、模型组、中药组及阴性对照组,每组8只.造模成功后,中药组及阴性对照组大鼠尾静脉注射相应外泌体(0.3 mg/kg),对照组及模型组注射等量磷酸盐(PBS),隔日1次,连续2周.生物机能实验系统记录大鼠胃电活动,ELISA检测大鼠血浆胃动素(MLT)含量,Western Blot法检测大鼠胃组织缝隙连接蛋白(CX43)蛋白表达,免疫荧光双染法检测酪氨酸激酶受体/平滑肌肌球蛋白重链(c-kit/SMMHC)、酪氨酸激酶受体/α平滑肌肌动蛋白(c-kit/α-SMA)共表达.结果 与对照组比较,模型组胃电慢波频率、主频率减低(P＜0.01),血浆中MLT含量减少(P＜0.01),胃窦组织中CX43、c-kit、α-SMA蛋白表达减低(P＜0.01,P＜0.05),SMMHC蛋白表达增高(P＜0.01);与模型组比较,中药组大鼠胃电慢波频率、主频率增高(P＜0.01),血浆中MLT含量增多(P＜0.01),胃窦组织中CX43、c-kit蛋白表达增高(P＜0.01,P＜0.05),SMMHC蛋白表达减低(P＜0.01).结论 半夏泻心汤血浆源性外泌体对胃电节律失常大鼠有一定干预作用,其对胃运动的调节可能通过促进c-kit及CX43蛋白表达,进而维持胃窦组织ICCs网络结构动态平衡实现.
Objective To observe the intervention effect of Banxia Xiexin Decoction(BXXXD)plasma-derived exosomes on the network structure of interstitial cells of Cajal(ICCs)in gastric antrum tissues of rats with gastric electric dysrhythmia and the possible mechanism of regulating gastric motility.Methods A total of 52 rats were selected,among them,20 rats were randomly divided into Chinese herb group and negative control group.The rat model of gastric electric dysrhythmia was established by"dieting every other day and drinking dilute hydrochloric acid"method,and BXXXD and distilled water were given for intervention after successful establishing of the rat model.After 2 weeks,plasma-derived exosomes were separated and identified.The remaining rats were randomly divided into control group,model group,Chinese herb group and negative control group,with 8 rats in each group.The exosome groups were injected with the corresponding exosomes(0.3 mg/kg)via the tail vein,and the control group and the model group were injected with the equal volume of phosphate buffer solution(PBS)once every other day for 2 weeks after successful modeling.After the intervention,the gastric electrical activity of rats was recorded by the biological function experiment system.The plasma motilin(MLT)content of rats was detected by ELISA.The expression of connexin 43(CX43)protein in gastric tissues of rats was detected by Western Blot.The co-expression of c-kit/SMMHC and c-kit/α-SMA were detected by immunofluorescence double staining.Results Compared with the control group,the frequency of gastric slow wave and the main frequency decreased(P＜0.01),the content of motilin in plasma decreased(P＜0.01),the expression of CX43,c-kit,and α-SMA proteins in gastric antrum tissues decreased(P＜0.01,P＜0.05),and the expression of SMMHC protein increased(P＜0.01)in the model group.Compared with the model group,the frequency of gastric slow wave and the main frequency in Chinese herb group increased(P＜0.01),the content of motilin in plasma increased(P＜0.01),the expression of CX43 and c-kit proteins in gastric antrum tissues increased(P＜0.05,P＜0.01),and the expression of SMMHC protein decreased(P＜0.01).Conclusions The plasma-derived exosomes of BXXXD have a certain intervenient effect on rats with gastric electric dysrhythmia.The regulation of gastric motility may be achieved by promoting the expression of c-kit and CX43 proteins,thereby maintaining the dynamic balance of ICCs network structure in gastric antrum tissues.
张迪;李雨静;盛春瑞;吉静;张冬梅;陈萌
北京中医药大学中医学院(北京 100029)北京中医药大学东直门医院科技处(北京 100700)
半夏泻心汤;外泌体;胃运动;缝隙连接蛋白43;Cajal间质细;中药复方;中西医结合
Banxia Xiexin Decoction;exosomes;gastric motility;connexin 43;interstitial cells of Cajal;Chinese herbal compound;integrative medicine
《中国中西医结合杂志》 2024 (004)
440-447 / 8
国家自然科学基金面上项目(No.82174270)
10.7661/j.cjim.20240224.034
评论