大黄素对洛哌丁胺致小鼠便秘的治疗作用

Therapeutic effect of emodin on constipation induced by loperamide in mice

目的:探讨大黄素对洛哌丁胺(loperamide,Lop)所致小鼠便秘的治疗作用及其潜在机制。方法:采用Lop构建小鼠便秘模型,采用大黄素处理便秘小鼠。统计小鼠在2 h观察期内的排便频率,收集粪便并检测粪便含水量;通过钡餐法检测肠通过时间;通过一氧化氮(nitric oxide,NO)检测试剂盒检测血清NO含量;通过HE染色观察小鼠结肠组织形态学改变;通过免疫组化检测小鼠结肠中血管活性肠肽受体1(vasoactive intestinal peptide receptor 1,VIPR1)和5-羟色胺(5-hydroxytryptamine,5-HT)4型受体(5-HT 4受体)的表达水平;通过Western blot检测瞬时受体电位阳离子通道亚家族V成员1(transient receptor potential cation channel subfamily V member 1,TRPV1)、胶质细胞源性神经营养因子(glial cell-derived neurotrophic factor,GDNF)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、一氧化氮合酶(nitric oxide synthase,NOS)、c-Kit和干细胞因子(stem cell factor,SCF)的表达水平。结果:大黄素可显著增加便秘小鼠在观察期内的排便次数以及粪便含水量(P<0.05或P<0.01),显著降低粪便的肠通过时间和血清NO水平(P<0.01)。HE染色结果显示,大黄素可减轻Lop引起的结肠组织炎症渗透。免疫组化结果显示,大黄素显著降低便秘小鼠结肠组织中VIPR1表达水平(P<0.01),显著增加5-HT 4受体表达水平(P<0.01)。Western blot结果显示,大黄素可显著降低小鼠结肠组织中TRPV 1和NOS的表达水平(P<0.01),显著增加GDNF、BDNF、c-Kit和SCF的表达水平(P<0.05或P<0.01)。结论:大黄素通过修复肠神经系统失调进而缓解Lop诱导的便秘。

AIM:To study the therapeutic effect of emodin on loperamide-induced constipation in mice.METHODS:The constipation model of mice was established by lopebutamine treatment,and the effects of emodin on defecation frequency,fecal water content and intestinal transit time of the mice during the observation period were detected.Inflammatory infiltration in colon tissue of the mice was observed by HE staining.Serum nitric oxide(NO)content was detected by commercially available kit.The expression of vasoactive intestinal peptide receptor 1(VIPR1)and 5-hydroxytryptamine type 4 receptor(5-HT 4 receptor)in mouse colon was determined by immunohistochemistry.The effects of emodin on the expression of transient receptor potential cation channel subfamily V member 1(TRPV1),glial cell-derived neurotrophic factor(GDNF),brain-derived neurotrophic factor(BDNF),nitric oxide synthase(NOS),c-Kit and their ligand stem cell factor(SCF)were detected by Western blot.RESULTS:Emodin significantly increased the number of defecation and fecal water content in the mice during the observation period,and significantly reduced the intestinal transit time and serum NO level in the mice(P<0.05).The results of HE staining showed that emodin significantly reduced the infiltration of colonic inflammation induced by loperamide.Emodin can significantly reduce the increase in VIPR1 induced by loperamine and increased the expression of 5-HT 4 receptor(P<0.01).Emodin increased the expression levels of GDNF and BDNF,reduced the expression levels of TRPV1 and NOS in the colon tissues of loperamine-induced constipation in mice,and significantly increased the expression of c-Kit and SCF(P<0.05 or P<0.01).CONCLUSION:Emodin promotes the defecation behavior of mice with loperamine-induced constipation by increasing intestinal peristalsis and activating a series of smooth muscle contraction-related factors.