痛泻要方对肠易激综合征内脏高敏性大鼠结肠组织肥大细胞活化、P物质表达及相关性的影响

Effect of Tongxie Yaofang on mast cells activation,expression of SP and their correlation on colon tissue of IBS visceral hypersensitivity rats

目的:观察肠易激综合征(IBS)内脏高敏性大鼠结肠肥大细胞(MC)、P物质(SP)及SP mRNA的改变及其相互关系,探讨MC活化、MC与SP表达的相关性在内脏高敏性发病机制的作用及痛泻要方对其干预作用。方法:采用乳鼠直肠扩张刺激法结合乳鼠与母鼠短暂分离制备IBS内脏高敏性大鼠模型。第12周-14周,灌胃痛泻要方2.03g/kg。第14周取结肠组织,采用甲苯胺蓝染色法对MC进行染色,以免疫组化染色法和RT-PCR分析结肠组织中SP、SP mRNA的表达变化,并以免疫组化染色法和甲苯胺蓝染色法进行SP、MC双重复染法,观察痛泻要方对结肠组织MC形态学、密度及SP、SP mRNA表达的影响,并通过Pearson相关系数、Kendall,s tau-b相关系数分析痛泻要方对MC、SP相关性的影响。结果:IBS内脏高敏性大鼠结肠中MC颗粒增多,SP、SP mRNA表达显著增高(P<0.01),MC周边SP表达显著增强,二者有明显相关性(P<0.01),经痛泻要方治疗后,结肠中MC形态改善、数量显著降低,SP、SP mRNA表达显著降低(P<0.01),MC与SP相关性仍明显(P<0.01)。结论:内脏高敏性大鼠肠道中MC与SP有明显相关性,痛泻要方可改善MC活化程度,降低SP释放,调控结肠SP、SP mRNA表达,通过调控神经介质和免疫细胞关系,而降低IBS内脏高敏性。

Objective： To observe the changes of mast cells, SP and SP mRNA and their correlation on colon tissue of IBS visceral hypersensitivity rats, to investigate the effect on the pathogenesis of visceral hypersensitivity of MC activation, correlation between the expression of MC and SP, and the intervention effect of Tongxie Yaofang on them. Methods： The IBS model of visceral hypersensitivity rats were established by colorectal distention stimulating baby rats combined with baby rats and female rats temporary separation. From 12th to 14th weeks, the rats were administered intragastrically Tongxie Yaofang solution 2.03g/kg. At the 14th week, the colon tissues were taken out, toluidine blue staining were stained for MC, the changes of expression of SP and SP mRNA were analyzed by immunohistochemistry staining methods and RT-PCR, and double counterstaining method that immunohistochemical method and toluidine blue staining method of MC and SP. To observe the effect of Tongxie Yaofang on the morphology and density of MC and the expression of SP and SP mRNA on colon tissue, and analyzed the effect of Tongxie Yaofang on correlation of MC and SP by Pearson correlation coefficient and Kendall＇s tau-b correlation coefficient. Results： On colon tissue of IBS visceral hypersensitivity rats, MC particles were increased, the expression of SP and SP mRNA were increased （P〈0.01）, the expression of SP around MC was enhanced, and there was obvious correlation （P〈0.01）. After treatment by Tongxie Yaofang, the morphology of MC was improved, the quantity of MC was decreased, and the expression of SP and SP mRNA was reduced （P〈0.01）, the correlation between MC and SP was significantly （P〈0.01）. Conclusion： The Correlation between MC and SP was significant on colon tissue of IBS visceral hypersensitivity rats, Tongxie Yaofang can improve degree of MC activation and reduce SP release and regulate the expression of SP and SP mRNA. And Tongxie Yaofang can reduce IBS visceral hypersensitivity by regulating correlation between and immune cells.