艾灸干预腹泻型肠易激综合征大鼠miR-24/SERT/5-HT通路改善内脏高敏感状态实验观察  

Moxibustion Improves Visceral Hypersensitivity in Rats with Diarrhea-predominant Irritable Bowel Syndrome by Exerting an Intervention Effect on the microRNA-24/Serotonin Transporter/5-Hydroxytryptamine Pathway

作　　者：廖路敏 王娇娇 储浩然[2] 李奎武 邹玲 阮静茹 祝姗姗 陈进雨 丁义侠[2] 王婧吉[2] 何雨霞 LIAO Lu-min;WANG Jiao-jiao;CHU Hao-ran;LI Kui-wu;ZOU Ling;RUAN Jing-ru;ZHU Shan-shan;CHEN Jin-yu;DING Yi-xia;WANG Jing-ji;HE Yu-xia(Anhui University of Chinese Medicine,Anhui Hefei 230012,China;The Second Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230061,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽中医药大学第二附属医院,安徽合肥230061

出　　处：《安徽中医药大学学报》2022年第5期59-66,共8页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(81774399);安徽省名老中医储浩然工作室建设项目(卫办秘[2015]404号)。

摘　　要：目的观察艾灸干预对腹泻型肠易激综合征(diarrhea predominant irritable bowel syndrome,IBS-D)大鼠微小RNA-24(micro RNA-24,miR-24)/5-羟色胺转运体(serotonin transporter,SERT)/5-羟色胺(5-hydroxytryptamine,5-HT)通路的影响,探究艾灸改善IBS-D大鼠内脏敏感性的作用机制。方法将SD大鼠随机分为正常组、模型组、艾灸组共3组,每组12只。采用母子分离+醋酸刺激+慢性束缚法制备IBS-D大鼠模型。艾灸组给予艾灸双侧“天枢”“上巨虚”20 min,连续7 d。分别于34、45、53 d检测各组大鼠体质量、稀便率和腹部回撤反射(abdominal withdrawal reflex,AWR)的最小容量阈值;苏木精-伊红染色检测结肠组织病理学变化;ELISA法测定大鼠血清、中脑、结肠组织中5-HT、SERT、5-羟色胺受体4(serotonin receptor 4,5-HT4R)、降钙素基因相关肽(calcitonin gene related peptide,CGRP)、P物质(substance P,SP)水平的变化;Western blot、RT-PCR检测大鼠中脑、结肠组织中SERT、5-HT4R、CGRP、SP的蛋白及mRNA表达水平;RT-PCR检测中脑、结肠组织中miR-24表达水平。结果与正常组比较,模型组大鼠稀便率升高(P<0.05)、AWR最小容量阈值下降(P<0.05);血清、中脑、结肠组织中5-HT、5-HT4R、CGRP、SP水平均显著升高(P<0.05),SERT水平均显著降低(P<0.05);miR-24表达水平显著升高(P<0.05),SERT蛋白及mRNA表达水平均明显降低(P<0.05),5-HT4R、CGRP、SP的蛋白及mRNA表达水平均明显升高(P<0.05);与模型组比较,艾灸组大鼠稀便率下降(P<0.05),AWR最小容量阈值上升(P<0.05);血清、中脑和结肠组织中5-HT、5-HT4R、CGRP、SP水平均显著降低(P<0.05),SERT水平显著升高(P<0.05);miR-24表达水平显著降低(P<0.05),SERT的蛋白及mRNA表达水平显著升高(P<0.05),5-HT4R、CGRP、SP的蛋白及mRNA表达水平显著降低(P<0.05)。结论艾灸干预改善IBS-D大鼠内脏高敏感性可能与下调miR-24表达,促进SERT对5-HT的重摄取,降低5-HT水平有关。Objective To investigate the mechanism of action of moxibustion in improving visceral hypersensitivity in rats with diarrhea-predominant irritable bowel syndrome(IBS-D)by observing the intervention effect of moxibustion on the microRNA-24(miR-24)/serotonin transporter(SERT)/5-hydroxytryptamine(5-HT)pathway.Methods Sprague-Dawley rats were randomly divided into normal group,model group,and moxibustion group,with 12 rats in each group.A rat model of IBS-D was established by mother-child separation combined with acetic acid stimulation and chronic restraint stress.The rats in the moxibustion group were given moxibustion at"Tianshu"and"Shangjuxu"at both sides for 20 minutes,once a day for 7 consecutive days.Body weight,loose stool rate,and minimum volume threshold of abdominal withdrawal reflex(AWR)were measured on days 34,45,and 53;HE staining was used to observe the histopathological changes of the colon;ELISA was used to measure the changes in the levels of 5-HT,SERT,serotonin receptor 4(5-HT4 R),calcitonin gene-related peptide(CGRP),and substance P(SP)in serum,midbrain,and colon tissue;Western blot and RT-PCR were used to measure the protein and mRNA expression levels of SERT,5-HT4 R,CGRP,and SP in the midbrain and colon tissue,and RT-PCR was used to measure the expression level of miR-24 in the midbrain and colon tissue.Results Compared with the normal group,the model group had a significant increase in loose stool rate(P<0.05),a significant reduction in the minimum volume threshold of AWR(P<0.05),significant increases in the levels of 5-HT,5-HT4 R,CGRP,and SP(P<0.05)and a significant reduction in the level of SERT in serum,midbrain,and colon tissue(P<0.05),a significant increase in the expression level of miR-24(P<0.05),significant reductions in the protein and mRNA expression levels of SERT(P<0.05),and significant increases in the protein and mRNA expression levels of 5-HT4 R,CGRP,and SP(P<0.05).Compared with the model group,the moxibustion group had a significant reduction in loose stool rate(P<0.05),a signif

关 键 词：艾灸 肠易激综合征大鼠 miR-24 5-羟色胺转运体 5-羟色胺 

分 类 号：R574[医药卫生—消化系统]