槐黄丸对慢传输型便秘大鼠结肠神经递质及SCF/c-kit通路的影响

Effect of Huaihuang Pills on Colonic Neurotransmitter and SCF/c-kit Pathway in Rats with Slow Transit Constipation

目的 探讨槐黄丸对慢传输型便秘(STC)大鼠的作用及其对结肠组织神经递质和干细胞因子/酪氨酸激酶受体(stem cell factor/kit proto-oncogene,SCF/c-kit)通路的影响。方法 将60只SD大鼠随机分为正常组、模型组、槐黄丸低剂量组(30 mg·kg^(-1))、槐黄丸中剂量组(75 mg·kg^(-1))、槐黄丸高剂量组(150 mg·kg^(-1))、莫沙必利组(2 mg·kg^(-1)),每组10只。除正常组外,其他组别均对大鼠进行大黄灌胃处理模拟STC造模,并以相应药物浓度给药。给药2周后,测量24 h首次排便时间、6 h排便粒数及粪便含水量、干重和肠道推进率。采用酶联免疫反应法(enzyme linked immunosorbent assay,ELISA)检测大鼠结肠P物质(Substance P,SP)、乙酰胆碱(acetylcholine,Ach)、一氧化氮(Nitric Oxide,NO)、血管活性肽(vasoactive intestinal polypeptide,VIP)的含量。RT-PCR法检测大鼠结肠c-kit和SCF的mRNA水平。Western blot检测大鼠结肠c-kit和SCF的蛋白水平。结果 与正常组比较,模型组大鼠首次排便时间明显延长,排便粒数及干重、粪便含水量明显减少,肠道推进率降低,SP、Ach含量降低,VIP、NO含量增高,SCF和c-kit的表达降低,差异均有统计学意义(P<0.05)。与模型组相比,槐黄丸低、中、高剂量组能缩短首次排便时间,6 h排便粒数及干重、粪便含水量明显增高,肠道推进率增高,差异均有统计学意义(P<0.05)。与槐黄丸低、中剂量组相比,槐黄丸高剂量组缓解STC的效果最佳,差异有统计学意义(P<0.05)。结论 槐黄丸对慢传输型便秘大鼠有明显的改善作用,其机制可能与调控结肠神经递质,提高SCF/c-kit信号通路有关。

Objective To investigate the effect of Huaihuang Pill on slow transit constipation(STC) rats and itsmechanism via colonic neurotransmitter and(stem cell factor) SCF/(kit proto-oncogene) c-kit pathway.Methods SixtySD rats were randomly divided into normal group, model group, Huaihuang Pill low-dose group(30 mg·kg^(-1)), HuaihuangPill medium-dose group(75 mg·kg^(-1)), and Huaihuang Pill high-dose group(150 mg·kg^(-1)), mosapride group(2 mg·kg^(-1)),10 rats in each group. Except for the normal group, rats in other groups were treated with rhubarb gavage to simulate STCmodeling, and they were administered according to the drug concentration of the corresponding group. After 2 weeks ofadministration, the time of first defecation in 24 hours, the number and dry weight of stool in 6 hours, the water content offeces, and intestinal propulsion rate were measured. ELISA method was used to detect the contents of in rat colon. RT-PCR method was used to detect the m RNA levels of c-kit and SCF in rat colon. The protein levels of c-kit and SCF inrat colon were determined by Western blot.Results Compared with the normal group, the rats in the model group hadsignificantly longer first defecation time. The number of stools, dry weight, and fecal water content in 6 hours were significantly reduced, and the intestinal propulsion rate was reduced. The content of SP and Ach were reduced, and thecontent of VIP and NO was increased. The expression of c-kit and SCF decreased, and the difference was statisticallysignificant(P<0.05). Compared with the model group, the low, medium, and high dose groups of Huaihuang Pills couldshorten the first defecation time, the number of stools and dry weight in 6 hours;and the water content of stool weresignificantly increased;the intestinal propulsion rate was increased;the SP and Ach content were increased;and the VIPand NO were reduced;the difference was statistically significant(P<0.05). Compared with the low and medium dosegroups of Huaihuang Pills, the high dose group of Huaihuang Pills had the best effect in relieving slow transitconstipation, and the difference was statistically significant(P<0.05).Conclusion Huaihuang Pills have a significantimprovement effect on rats with slow transit constipation, and its mechanism may be related to the regulation of colonicneurotransmitters and the improvement of SCF/c-kit signaling.