川陈皮素对慢传输型便秘小鼠的治疗作用及机制研究

Therapeutical Effects and Its Mechanism of Nobiletin on Slow Transit Constipation Mice

目的探讨川陈皮素(NOB)对慢传输型便秘(STC)小鼠的治疗作用及其机制。方法随机将小鼠分为正常组、模型组、阳性对照组(莫沙必利,2.5 mg·kg^(-1))及NOB低、中、高剂量组(10、20、40 mg·kg^(-1)),每组12只。模型组和给药组均采用盐酸洛哌丁胺诱导STC小鼠模型,造模成功后给予相应药物灌胃处理,每天1次,共2周。检测各组小鼠24 h内排便量、粪便含水量、肠道推进率及结肠肌电变化;HE染色观察各组小鼠结肠组织病理学变化;ELISA法检测各组小鼠结肠组织中5-羟色胺(5-HT)、血管活性肽(VIP)、一氧化氮(NO)含量及一氧化氮合酶(NOS)活性;Western blot检测各组小鼠结肠组织中干细胞因子(SCF)和酪氨酸激酶受体(c-kit)蛋白表达水平。结果与正常组比较,模型组小鼠结肠损伤明显,24 h排便量、粪便含水量、肠道推进率、结肠肌电慢波振幅及结肠组织中SCF和c-kit蛋白表达均显著降低(P<0.05),而结肠肌电慢波频率、结肠组织中5-HT、VIP、NO和NOS水平显著升高(P<0.05);与模型组比较,NOB中、高剂量组及阳性对照组小鼠结肠损伤减轻,24 h内排便量、肠道推进率、结肠肌电慢波振幅及结肠组织中SCF和c-kit蛋白表达均显著升高(P<0.05),而结肠肌电慢波频率及结肠组织中5-HT、VIP、NO和NOS水平均显著降低(P<0.05),而NOB低剂量组以上指标无显著性差异(P>0.05)。结论NOB可通过调节结肠肌电、增强肠道神经传递、促进肠道推进,从而改善STC小鼠排便情况,其作用机制可能与上调SCF和c-kit蛋白表达有关。

Objective To investigate the therapeutical effect and mechanism of nobiletin(NOB)on slow transport constipation(STC)mice.Methods Mice were randomly divided into normal group,model group,positive control group(mosapride,2.5 mg·kg^(-1))and NOB low-dose,medium-dose and high-dose groups(10,20,40 mg·kg^(-1)),with 12 mice in each group.The model group and the administration groups were then modeled using loperamide hydrochloride intragastrically to obtain slow transit constipation,and each group was given corresponding drugs by intragastric administration once a day for 2 weeks after successful modeling.24 h defecation grain number,water content of faeces,changes of intestinal transit rate and colonic myoelectric of mice in each group were detected.Contents of 5-hydroxytryptamine(5-HT),vasoactive peptide(VIP)and nitric oxide(NO)and the activity of nitric oxide synthase(NOS)in colon tissues of mice in each group were determined by ELISA.Western blot was used to detect stem cell factor(SCF)and tyrosine kinase receptor(c-kit)protein expression levels in colon tissues of mice in each group.Results Compared with normal group,colon injury was obvious in model group,24 h defecation grain number,water content of faeces,intestinal propulsion rate,colonic myoelectric slow wave amplitude,SCF and c-kit protein expression in colon tssue were significantly decreased(P<0.05),while colonic myoelectric slow wave frequency,the levels of colon 5-HT,VIP,NO and NOS were significantly increased(P<0.05).Compared with model group,colon injury was alleviated in medium,high dose NOB groups and positive control group,24 h defecation grain number,water content of faeces,intestinal propulsion rate,colonic myoelectric slow wave amplitude,SCF and c-kit protein expression in colon tssue were significantly increased(P<0.05),while colonic myoelectric slow wave frequency,the levels of colon 5-HT,VIP,NO and NOS were significantly decreased(P<0.05),but there were no significant differences in the above indexes in NOB lowdose group(P>0.05).Conclusion NOB can improve the defecation of STC mice by regulating colonic myoelectric,enhancing intestinal neurotransmission and promoting intestinal propulsion,and its mechanism may be related to the upregulation of SCF and c-kit protein expression.