不同刺灸法对功能性便秘大鼠血浆SP含量、结肠SP mRNA表达的影响研究

摘要

目的:探讨不同刺灸方法对功能性便秘大鼠血浆P物质(substance P,SP)含量、结肠SP mRNA表达的影响.方法:将60只健康雄性SD大鼠按随机数字表法分为空白对照组(8只)、模型组(1 1只)、药物对照组(8只)、毫针组(1 1只)、电针组(1 1只)和艾灸组(1 1只).造模同时空白组和模型组不予干预,药物对照组予西沙必利混悬液灌胃,电针组、毫针组、艾灸组则分别给予电针、毫针和艾灸治疗6天.治疗结束后观察各组大鼠24 h粪便量和粪便含水量,结肠组织结构及粘膜层酸性粘液,血浆SP含量,结肠SP mRNA表达.结果:与空白对照组比较,模型组24 h粪便量及粪便含水量显著下降(P＜0.01);与模型组比较,各组大鼠24 h粪便量均增加(P＜0.05或0.01),除艾灸组外,其余各组大鼠24 h粪便量含水量也明显增加(P＜0.01);与药物组比较,艾灸组24 h粪便量含水量减少(P＜0.01);不同刺灸法干预组之间比较,24小时粪便量各组无差异(P＞0.05),24小时粪便含水量毫针组、电针组均高于艾灸组(P＜0.01),毫针组和电针组无差异(P＞0.05).大鼠结肠组织HE染色显示结构正常、完整,各组间无明显差异.采用PAS染色显示酸性粘液层,与空白组比较,模型组酸性粘液明显减少;与模型组比较,药物组、毫针组、电针组、艾灸组均有不同程度增加.与空白对照组比较,模型组血浆SP含量下降(P＜0.05);与模型组比较,毫针组、艾灸组含量均升高(P＜0.01);与药物组比较,三个刺灸法干预组合量均高于药物组(P＜0.05或0.01);不同刺灸法干预组之间比较,毫针组高于艾灸组及电针组(P＜0.05),艾灸组与电针组无差异.与空白对照组比较,模型组结肠组织SP mRNA的表达升高(P＜0.05);与模型组比较,四个干预组的结肠组织SP mRNA表达均下降(P＜0.05);与药物组比较,毫针组、电针组、艾灸组无差异(P＞0.1);不同刺灸法干预组之间比较,艾灸组低于毫针组(P＜0.05).结论:不同刺灸方法治疗功能性便秘的疗效不同可能与调节血浆SP含量、结肠组织SP mRNA表达的机制有关.%Objective To explore the effects of different acupuncture-moxibustion methods on the plasma substance P (SP) content,and the expression of colonic SP mRNA of the rats with functional constipation.Methods Sixty healthy male SD rats were divided into six groups according to the random number table,including blank control group (group B,n=8),model group (group M,n=11),drug control group (group D,n=8),filiform needle group (group F,n=11),electroacupuncture group (group E,n=11) and moxibustion group (group MB,n=11).The rats in group B and group M accepted no intervention,gavage with cisapride suspension was conducted in group D,and electroacupuncture,filiform needling and moxibustion therapies were carried out in group E,group F and group MB for 6 days.After treatment,the stool quantity and fecal moisture content during 24 h of the rats were observed in each group,colon tissue structure and the acidic mucus in mucosa layer were observed,plasma SP content was detected and the expression of colonic SP mRNA was detected.Results compared with group B,the stool quantity and fecal moisture content during 24 h of the rats in group M decreased significantly (P＜0.01);compared with group M,the stool quantity during 24 h of the rats in each group increased (P＜0.05 or 0.01),and the fecal moisture content during 24 h of the rats in each group also increased significantly except group MB (P＜0.01);compared with group D,the fecal moisture content during 24 h of the rats in group MB decreased (P＜0.01);according to the comparison among the groups with different acupuncture-moxibustion methods,there was no difference in the stool quantity during 24 h of the rats in each group (P＞0.05),the fecal moisture contents during 24 h of the rats in group F and group E were higher than that of group MB (P＜0.01),and there was no difference between group F and group E (P＞0.05).HE staining of colon tissue of rats showed that the structure was normal and complete,and there was no difference among each group.PAS staining showed that the acidic mucus level of the rats in group M decreased significantly when compared with group B;compared with group M,the acidic mucus level of the rats in group D,fgroup F,group E and group MB increased in varying degrees.Compared with group B,the plasma SP content of the rats in group M decreased (P＜0.05);compared with group M,the plasma SP contents of the rats in group F and group MB increased (P＜0.01);the plasma SP contents of the rats in group F,group MB and group E were higher than that of group D (P＜0.05 or 0.01);according to the comparison among the groups with different acupuncture-moxibustion methods,the plasma SP content of the rats in group F was higher than that of group MB and electro-acupuncture group (P＜0.05),and there was no difference between group MB and group E.Compared with group B,the expression level of colonic SP mRNA of the rats in group M elevated (P＜0.05);compared with group M,the expression level of colonic SP mRNA of the rats in the four intervention groups declined (P＜0.05);compared with drug group D,there was no difference among group F,group E and group MB (P＞0.1);according to the comparison among the groups with different intervention methods,the expression level of colonic SP mRNA of the rats in group MB was lower than that of group F (P＜0.05).Conclusion The different efficacy of different acupuncture-moxibustion methods in treatment of functional constipation may be related with the mechanism of regulation of plasma SP content and the expression level of colonic SP mRNA.