腹部推拿联合5-HT4受体激动剂调节SCF/c-kit信号通路对STC大鼠的作用研究

Effects of abdominal massage combined with 5-HT4 receptor agonist on SCF/c kit signaling pathway in STC rats

目的观察腹部推拿联合5-HT4受体激动剂调节SCF/c-kit信号通路对慢传输型便秘(STC)大鼠的作用。方法2017年10月—2018年6月于新疆维吾尔自治区中医药研究院进行实验。65只雄性大鼠随机数字表法分为5组,对照组、模型组、推拿组、莫沙必利组、推拿联合莫沙必利组(联合组),每组13只。除对照组外,其余各组采用肠神经节消融术建立STC大鼠模型,推拿组进行腹部推拿,莫沙必利组大鼠予0.15 mg/ml莫沙必利10 ml/kg灌胃,联合组予推拿+相同浓度的莫沙必利灌胃,各组干预方式每日1次,连续14 d。实验结束后比较各组大鼠体质量、粪便含水率、血浆SP、NO、5-HT及VIP水平、肠推动率,实时定量PCR检测大鼠结肠组织中5-HT3R和5-HT4R mRNA的表达,Western-blot检测大鼠结肠组织中SCF、c-kit蛋白水平。结果与对照组比较,模型组大鼠体质量增加值显著升高(F=11910.958,P=0.021),粪便含水率显著降低(F=81.938,P=0.018),血浆SP水平显著降低,NO、5-HT及VIP水平均显著升高(F/P=621.651/0.011、4298.343/0.008、2282.876/0.013、834.829/0.026),肠推动率显著降低(P<0.05),结肠组织中5-HT3R及5-HT4R mRNA水平、SCF、c-kit水平均显著下降(P<0.05)。与模型组比较,推拿组、莫沙必利组、联合组大鼠体质量增加值均显著降低(P<0.05),粪便含水率显著增加(P<0.05),血浆SP水平显著升高,NO、5-HT及VIP水平均显著降低(P<0.05),肠推动率均显著升高(P<0.05)。结肠组织中5-HT3R及5-HT4R mRNA水平、SCF、c-kit蛋白水平均显著升高(P<0.05),且联合组大鼠各项指标转归效果均显著优于推拿组及莫沙必利组(P<0.05)。结论腹部推拿联合5-HT4受体激动剂能够调节STC大鼠体内SCF/c-kit信号通路,缓解STC大鼠疾病状态,改善其疾病转归。

Objective To observe the effect of abdominal massage combined with 5-HT4 receptor agonist on SCF/c kit signal pathway in slow transit constipation(STC)rats.Methods The experiment was conducted in Xinjiang Uygur Autonomous Region Institute of Traditional Chinese Medicine from October 2017 to June 2018.Sixty-five male rats were randomly divided into 5 groups:control group,model group,massage group,mosapride group,massage combined with Mosapride group(combined group).In addition to the control group,the other groups used intestinal ganglia ablation to establish STC rat model.The rats in mosapride group were given 0.15 mg/ml mosapride 10 ml/kg intragastric administration,and the rats in combination group were given massage+mosapride of the same concentration intragastric administration.Each group was intervened once a day for 14 days.At the end of the experiment,the body weight,fecal water content,plasma SP,no,5-HT and VIP levels,intestinal motility were compared.The expression of 5-HT3R and 5-HT4R mRNA was detected by real-time quantitative PCR,and SCF and c-kit protein levels were detected by Western blot.Results Compared with the control group,the body weight increment of the model group was significantly higher(F=11910.958,P=0.021),and the fecal water content was significantly lower(F=81.938,P=0.018),the plasma SP level was significantly reduced,the levels of no,5-HT and VIP were significantly increased(F/P=621.651/0.011,4298.343/0.008,2282.876/0.013,834.829/0.026),the intestinal motility rate was significantly reduced(P<0.05),and the levels of 5-HT3R and 5-HT4R mRNA,SCF and C kit water in colon tissue were significantly decreased(P<0.05).Compared with the model group,the body weight of rats in massage group,mosapride group and combination group decreased significantly(P<0.05),fecal water content increased significantly(P<0.05),plasma SP level increased significantly,no,5-HT and VIP levels decreased significantly(P<0.05),and intestinal motility increased significantly(P<0.05).The levels of 5-HT3R and 5-HT4R mRNA,SCF and c-kit protein in colon tissue were significantly increased(P<0.05),and the outcome of the combined group was significantly better than that of the massage group and Mosapride group(P<0.05).Conclusion Abdominal massage combined with 5-HT4R receptor agonist can regulate SCF/c-kit signaling pathway in STC rats,alleviate the disease status and improve the disease outcome.