基于神经免疫调节途径探讨温胃阳汤对功能性消化不良大鼠的作用

Exploring the effect of Wenweiyang Decoction on rats with functional dyspepsia based on neuroimmunomodulatory pathway

目的观察温胃阳汤对功能性消化不良(FD)大鼠十二指肠肥大细胞(MC)、类胰蛋白酶(MCT)、组胺、神经胶质(S100)的影响,探讨温胃阳汤治疗FD的作用机制。方法将60只SD大鼠随机分为空白组、模型组、雷尼替丁组及温胃阳汤低、中、高剂量组,每组10只。除空白组外,其余组大鼠均采用番泻叶联合夹鼠尾法制备脾阳虚型FD模型。进行胃肠运动实验,观察各组大鼠胃排空率和小肠推进率;采用甲苯胺蓝染色观察FD大鼠MC及MC脱颗粒数目;ELISA检测FD大鼠十二指组织MCT、组胺的自发释放水平;采用免疫组化法检测十二指肠组织S100的平均光密度,S100免疫组织化学染色法观察神经纤维的分布及变化情况等。结果与空白组比较,模型组大鼠胃排空率和小肠推进率均降低(P<0.01),十二指肠黏膜MC明显增多且MC脱颗粒明显,MCT、组胺浓度均升高(P<0.01),钙离子浓度降低(P<0.01),十二指肠黏膜下层S100阳性神经纤维表达增强(P<0.01);与模型组比较,雷尼替丁组与温胃阳汤组大鼠胃排空率和小肠推进率均升高(P<0.05),温胃阳汤中、高剂量组大鼠十二指肠黏膜MC及脱颗粒明显减少,温胃阳汤低、中、高剂量组的MCT、组胺浓度均降低(P<0.05),雷尼替丁组及温胃阳汤组S100阳性神经纤维表达减弱(P<0.01),以及钙离子浓度升高(P<0.01)。结论温胃阳汤可明显促进FD大鼠胃肠运动功能,可能是通过抑制十二指肠MC活化和脱颗粒,从而抑制MC分泌组胺,下调MCT表达,激活神经细胞Ca2+内流,改变神经纤维分布来发挥机制。

Objective To observe the effects of Wenweiyang Decoction on duodenal mast cells(MC),trypsin-like(MCT),histamine and neuroglia(S100)in rats with functional dyspepsia(FD),and to explore the mechanism of Wenweiyang Decoction in the treatment of FD.Methods 60 SD rats were randomly divided into blank group(n=10),model group(n=10),ranitidine group(n=10)and Wenweiyang Decoction low,middle and high dose groups(n=10).Except for the blank group,the FD model of spleen Yang deficiency was established by Senna leaf combined with rat tail clamping method in other groups.Gastrointestinal motility test was performed to observe the gastric emptying rate and intestinal propulsion rate of rats in each group.Toluidine blue staining was used to observe the number of MC and MC degranulation in FD rats.ELISA was used to detect the spontaneous release of MCT and histamine in the twelve fingers of FD rats.The average optical density of S100 in duodenal tissue was detected by immunohistochemical method,and the distribution and changes of nerve fibers were observed by S100 immunohistochemical staining.Results Compared with the blank group,the gastric emptying rate and intestinal propulsion rate in the model group decreased,the duodenal mucosa MC significantly increased and MC degranulated significantly,the concentrations of MCT and histamine increased(P<0.01),the calcium concentration decreased(P<0.01),and the expression of S100 positive nerve fibers in duodenal submucosa increased(P<O.01).Compared with the model group,the gastric emptying rate and intestinal propulsion rate in the ranitidine group and Wenweiyang Decoction group increased,the MC and degranulation of duodenal mucosa in the middle and high dose groups decreased significantly,the concentrations of MCT and histamine in the low,middle and high dose groups decreased,and the expression of Si00 positive nerve fibers decreased in the ranitidine group and Wenweiyang Decoction group(P<0.01).And the concentration of calcium increased(P<0.01).Conclusion Wenweiyang Decoction can obviously promote gastrointestinal motility in FD rats,which may be due to inhibition of MC activation and degranulation in duodenum,inhibition of histamine secretion by MC,down-regulation of MCT expression,activation of Ca influx of nerve cells and change of nerve fiber distribution.