目的:探讨胃肠舒促胃肠动力作用机制。方法:原代培养胃肠平滑肌细胞,分胃肠舒大、中、小剂量组,西沙必利组和正常对照组,各组用相应受试药物干扰胃肠平滑肌细胞24h。提取细胞总RNA,应用逆转录-聚合酶链式反应(RT-PCR)法,半定量分析eNOS mRNA表达水平。结果:胃肠舒大、中剂量组与正常对照组比较,胃肠平滑肌细胞,eNOS mRNA表达水平显著降低(P〈0.01,P〈0.05)。结论:胃肠舒促胃肠动力的作用机制可能是通过抑制胃肠平滑肌细胞eNOS mRNA表达而抑制细胞内NO生成,进而抑制相关酶的活化,拮抗cGMP生成,使胞内Ca2+增多,Ca2+信号系统引发一系列生理功能而促进胃肠平滑肌收缩。
Objective: To investigate the mechanism of Weichangshu promots gastrointestinal motility.Methods: Primary culture gastrointestinal smooth muscle cells,experiment was divided into Weichangshu high-dose group,Weichangshu middle-dose group,Weichangshu low-dose group,Cisapride group,and normal control group,tested drugs were used in each group interfere with cell 24h,detect the release of NO using Flow cytometry.Then extract the total RNA of cells,use reverse transcription-polymerase chain reaction,semi-quantitative analysis the changes of the expression of eNOS mRNA levels in gastrointestinal smooth muscle cells.Results: Compared with the normal control group,fluorescence intensity of NO release significantly reduced(P〈0.01,P〈0.05),and eNOS mRNA expression was significantly lower(P〈0.01,P〈0.05) of gastrointestinal smooth muscle cells in Weichangshu high-dose group and the middle-dose group.Conclusion: The mechanism of Weichangshu gastrointestinal motility-promoting was may be inhibiting the release of intracellular NO by inhibiting the expression of eNOS mRNA in gastrointestinal smooth muscle cells,and then inhibiting the activation of enzymes,antagonized the formation of cGMP,thus increasing intracellular Ca2+,Ca2+ signaling system triggered a series of physiological functions,thereby promoting effects of gastrointestinal smooth muscle contraction.