清化理气通便法对湿热证功能性便秘大鼠脑肠肽的影响

The influences of the Qinghua Liqi Tongbian therapy on brain-gut peptide in rats with functional constipation of damp-heat syndrome

目的:通过观察清化理气通便法对湿热证功能性便秘(Functional Constipation,FC)大鼠脑肠肽促肾上腺皮质激素释放因子(Corticotropin Releasing Factor,CRF)、胆囊收缩素(Cholecystokinin,CCK)、一氧化氮(Nitric Oxide,NO)、诱导型一氧化氮合酶(Inducible Nitric Oxide Synthases,iNOS)表达水平的影响,探讨其治疗湿热证FC的机制。方法:将无特定病原体级Wistar雄性大鼠按照随机数字法分为空白组(8例)与造模组(40例),采用“复方地芬诺脂混悬液灌胃+潮湿环境+高脂高糖饮食”复合法建立湿热证FC大鼠模型,造模成功后,将造模组随机分为模型组,莫沙必利组以及中药低、中、高剂量组,每组8只。给予相应药物连续干预2周后,观察各组大鼠首粒黑便排出时间,实时荧光定量聚合酶链式反应(Polymerase Chain Reaction,PCR)法检测CCK、CRF mRNA表达情况,酶联免疫吸附剂测定(Enzyme-linked Immunosorbent Assay,ELISA)法检测血清NO、iNOS含量。结果:与空白组相比,模型组大鼠首粒黑便排出时间延长、结肠组织中CRF mRNA表达下调,CCK mRNA表达上调,血清NO、iNOS含量上升,差异均有统计学意义(P<0.05或P<0.01)。中药各剂量组及莫沙必利组黑便排出时间均改善,结肠组织中CRF mRNA表达升高,CCK mRNA表达下降,血清NO、iNOS含量降低,差异有统计学意义(P<0.05或P<0.01)。结论:清化理气通便法可有效改善湿热证FC大鼠排便时间延长,其机制可能与调控CCK、CRF、NO、iNOS等脑肠肽有关。

Objective:To explore the mechanism of treating damp-heat syndrome functional constipation by the Qinghua Liqi Tongbian therapy(清化理气通便法)according to observing its influences on the expression of CRF,CCK,NO,and iNOS in rats with functional constipation of damp-heat type.Methods:According to the random number method,SPF-grade Wistar male rats were divided into the blank group(8 rats)and the model group(40 rats).The damp-heat FC rat models were established by the composite method of“compound diphenoxylate suspension plus humid environment and high-fat-sugar diet”.After the model was successfully established,the model rats were randomly divided into the model group,the mosapride group,and TCM medicine low,medium,and high dose groups,8 rats in each group.After continuous intervention with corresponding drugs for 2 weeks,the time of the first black defecate of rats in each group was observed.The expressions of CCK and CRP mRNA were detected by quantitative-real-time PCR,and the serum NO and iNOS levels were detected by ELISA.Results:Compared with the blank group,the rats in the model group showed longer discharge time of the first black defecate,down-regulated CRF mRNA expression and up-regulated CCK mRNA expression in colon tissue,and increased serum NO and iNOS levels.The differences were statistically significant(P<0.05 or P<0.01).In TCM medicine low,medium,and high dose groups,and the mosapride group,the discharge time of black defecate was ameliorated,the expression level of CRF mRNA in colon tissue was increased,CCK mRNA level was decreased,and serum NO and iNOS levels were decreased.The differences were statistically significant(P<0.05 or P<0.01).Conclusion:The Qinghua Liqi Tongbian therapy can effectively ameliorate the defecation time of rats with damp-heat syndrome FC.The mechanism might be related to the regulation of CCK,CRF,NO,iNOS and other brain-gut peptides.