附子-肉桂对慢传输型便秘大鼠5-HT信号通路及c-Kit表达的影响

Effect of Fuzi-Rougui on 5-HT Signal Pathway and c-Kit Expression in Rats with Slow Transit Constipation

目的研究不同比例附子-肉桂配伍通过调控STC大鼠5-HT信号通路及c-Kit表达改善肠道动力的作用机制。方法运用盐酸洛哌丁胺灌胃建立STC大鼠模型,随机分为空白组、模型对照组、附子-肉桂(2∶1)组,附子-肉桂(1∶2)组、附子-肉桂(1∶1)组、莫沙必利组。各组按照对应药物或蒸馏水灌胃14天。观察大鼠一般情况,计算大鼠粪便粒数、肠道推进率,HE观察大鼠结肠病理改变,ELISA法测定大鼠血清、结肠5-HT含量,免疫荧光染色检测结肠CgA阳性表达,免疫组化法检测结肠5-HT3R/5-HT4R、c-Kit表达情况,RT-PCR法检测5-HT3R/5-HT4R mRNA表达水平。结果各组大鼠结肠无明显病理改变,与空白组相比,模型对照组大鼠粪便含水率、肠道推进率显著降低(P<0.01),5-HT含量明显下调,CgA、5-HT3R/5-HT4R蛋白与基因表达水平均显著下降(P<0.01),c-Kit表达减少。与模型组相比,附子-肉桂(2∶1)组、附子-肉桂(1∶1)组、莫沙必利组粪便粒数、肠道推进率、5-HT含量、c-Kit、CgA阳性表达显著增加(P<0.01),5-HT3R/5-HT4R蛋白与基因表达水平显著上调(P<0.01)。结论附子-肉桂2∶1及1∶1比例配伍能明显增加粪便粒数,提高肠推进率,其作用可能与通过调节结肠c-Kit、CgA表达,增加Cajal间质细胞数及5-HT含量,从而上调5-HT3R/5-HT4R表达有关。

Objective To study the promote colon motility mechanism of different proportions of Fuzi-Rouguiby 5-HT signal path and c-Kit in slow transit constipation.Methods The STC model was established by loperamide hydrochloride.The rats were divide to blank control group,model group,Fuzi-Rougui(2∶1)group,Fuzi-Rougui(1∶2)group,Fuzi-Rougui(1∶1)group and Mosapride group.The groups were given corresponding drugs.Calculation the intestinal transit rate and Fecal moisture content Before and after medication.5-HT content was determined by ELISA.The positive expression of CgA was determined by immunofluorescence staining.The expression of Cajal,5-HT3R/5-HT4R were detected by immunohistochemistry.The 5-HT3R/5-HT4R mRNA levels were detected by reverse transcription-polymerase chain reaction.Results The average value of intestinal transit rate,dejection amounts,5-HT content,CgA/5-HT3R/5-HT4R,c-Kit expression of model group was lower than blank control group.The average value of intestinal transit rate,dejection amounts,5-HT content,CgA/5-HT3R/5-HT4R,Cajal expression of Fuzi-Rougui(2∶1)group,Fuzi-Rougui(1∶1)group and Mosapride group were higher than model group.Conclusion Proportions 2∶1,1∶1 of Fuzi-Rouguican Promote Intestinal motility effectively.The increased level of Intestinal motility as well as elevated Up-regulating the content of 5-HT and the expression of CgA/5-HT3R/5-HT4R and c-Kit.