运脾调气法在腹泻型肠易激综合征大鼠中的应用分析及作用机制探究

Application and Mechanism of Activating Spleen and Regulating Qi Method in Diarrhea-Type Irritable Bowel Syndrome Rats

目的 探究运脾调气法在腹泻型肠易激综合征大鼠中的应用结局及作用机制,旨在为临床使用提供可循证的实验数据和可参考的理论依据。方法 选取64只成年清洁级(SPF)同月龄健康Wistar大鼠,采用随机数字表法将其分为正常组20只,其余44只大鼠予以造模;模型成功后,再次参考随机数字表法将其分为模型组和中药组,每组22只。正常组大鼠不予任何干预,模型组大鼠予以0.9%生理盐水10 mL/kg灌胃、1次/d、连续14 d,中药组予以具有运脾调气功能的中药汤剂灌胃、1次/d、连续14 d。观察和比较大鼠结肠组织病理变化、分析结肠组织跨膜蛋白toll样受体4抗体(TLR4)和核转录因子-κB(p65)抗体[NF-κB(p65)]mRNA及其蛋白表达水平和脑肠肽平均光密度值[P物质(SP)、血管活性肠肽(VIP)]。结果 模型组和中药组成模评定各处死2只,造模成功率100.00%。显微镜下可见正常组大鼠结肠组织黏膜结构完整、间质内无水肿及纤维结缔组织增生;模型组大鼠结肠组织黏膜存在局部炎性浸润,但其黏膜及黏膜下结构尚完整;而中药组炎性反应显著改善且结肠组织黏膜结构完整。3组大鼠结肠组织TLR 4 mRNA表达及蛋白灰度值比较,以模型组表达最高,其次为中药组,而正常组最低,其间差异有统计学意义(P<0.05);NF-κB(p65)mRNA表达及蛋白灰度值比较,以模型组表达最高且均高于正常组和中药组,差异有统计学意义(P<0.05),而正常组和中药组比较,差异无统计学意义(P>0.05)。3组大鼠脑、肠组织脑肠肽SP、VIP光密度值分别比较,以模型组最高且均高于正常组和中药组,差异有统计学意义(P<0.05);中药组与正常组比较差异无统计学意义(P>0.05)。结论 运运脾调气法通过健运脾胃、调气理肠等标本兼治的方式紧贴腹泻型肠易激综合征虚实夹杂的病机,达到了“正气存内、祛邪外出”等目的,具有光明的应用前景,为临床使用提供了依据。

Objective To investigate the outcome and mechanism of activating spleen movement and regulating Qi method in diarrhea-type irritable bowel syndrome(IBS) rats and provide the evidence-based experimental data and theoretical basis for clinical use.Methods A total of 64 adult clean grade(SPF) healthy Wistar rats at the same age were randomly divided into normal group(20 rats) by random number table method and the other 44 rats were modeled.After the model was successful, they were divided into model group and traditional Chinese medicine group again by referring to the random number table method, with 22 rats in each group.Rats in the normal group were given no intervention.Rats in the model group were given 0.9% normal saline 10 mL/kg by intragastric administration, once a day, for consecutive 14 days.Rats in the traditional Chinese medicine group were given traditional Chinese medicine decoction with the function of activating spleen and regulating Qi, once a day, for consecutive 14 days.The pathological changes of colon tissue in rats were observed, and the mRNA and protein expression levels of transmembrane protein Toll-like receptor 4 antibody(TLR4) and nuclear transcription factor-κB(p65) antibody[NF-κB(p65)] and the mean optical density of brain-gut peptide[substance P(SP) and vasoactive gut peptide(VIP)] were analyzed.Results Two rats were killed in the model group and the traditional Chinese medicine model respectively, and the success rate of the model group was 100.00%.Under the microscope, the normal group had intact mucosal structure, no edema and fibrous connective tissue hyperplasia in the interstitium.In model group, there was local inflammatory infiltration in colonic tissue mucosa, but the mucosa and submucosa structure were intact.The inflammatory response of the traditional Chinese medicine group was significantly improved and the mucosal structure of the colon tissue was intact.The expression of TLR 4 mRNA and protein gray level in the colon tissues of the model group were the highest, followed by those of the traditional Chinese medicine group, and thoseof the normal group were the lowest, with statistical significance(P<0.05).The mRNA expression and protein gray level of NF-κB(p65) in the model group were higher than those in the normal group and the traditional Chinese medicine group(P<0.05),but there was no significant difference between the normal group and the traditional Chinese medicine group(P>0.05).The substance P(SP) and VIP optical density values in brain and intestine tissues of rats in the three groups were compared, and the model group had the highest values, which were higher than those in normal group and traditional Chinese medicine group and the difference was statistically significant(P<0.05).There was no significant difference between the traditional Chinese medicine group and the normal group(P>0.05).Conclusion The method of activating spleen and regulating Qi is closely related to the pathogenesis of diarrhea-type IBS by means of improving spleen and stomach, regulating Qi and regulating intestines, and so on.It can achieve the purpose of “maintaining healthy Qi and removing pathogenic factors” and has a bright application prospect, providing a basis for clinical use.