基于干细胞因子(SCF)/c-kit信号通路探讨和胃理气方治疗功能性消化不良胃肠运动功能障碍的作用机制

Study on Mechanism of Stomach-Harmonizing and Qi-Soothing Recipe for Gastrointestinal Motility Dysfunction of Functional Dyspepsia Based on Stem Cell Factor/c-kit Signaling Pathway

【目的】观察和胃理气方对功能性消化不良肝郁模型大鼠胃肠动力及干细胞因子(SCF)/c-kit信号通路的影响,从分子水平探讨其作用机制。【方法】选取敞箱实验和糖水偏好实验评分相似的40只SPF级健康Wistar大鼠,随机分为正常组、模型组、中药组和西药组,每组10只。除正常组外,其他组别采用夹尾刺激+不规则喂养复合因素法建立功能性消化不良肝郁动物模型。成功造模后,中药组和西药组分别用和胃理气方和吗丁啉进行干预治疗。通过敞箱、糖水偏好实验评价大鼠抑郁状态;采用胃排空、小肠推进实验评价胃肠运动情况;分别用免疫组织化学、蛋白免疫印迹法检测大鼠胃窦、小肠组织中c-kit、SCF的蛋白表达水平;采用实时定量聚合酶联反应(RT-PCR)法检测大鼠胃窦、小肠黏膜组织中c-kit、SCF的mRNA表达水平。【结果】与正常组比较,模型组大鼠糖水消耗率下降,敞箱实验水平及垂直运动得分下降,胃排空及小肠推进减慢,胃窦、小肠组织中c-kit、SCF阳性表达细胞光密度值降低,c-kit、SCF的蛋白、mRNA表达水平均下降(均P<0.05)。与模型组比较,中药组和西药组糖水消耗率提高,敞箱实验水平及垂直运动得分增加,胃排空及小肠推进加快,胃窦、小肠组织中c-kit、SCF阳性细胞光密度值升高,c-kit、SCF的蛋白、mRNA表达水平均升高(均P<0.05),且中药组和西药组比较,上述指标均无统计学差异(P>0.05)。【结论】肝郁可致胃肠动力障碍,和胃理气方可以改善功能性消化不良肝郁大鼠抑郁状态,促进胃肠动力,其分子作用机制与上调SCF/c-kit信号通路相关蛋白表达有关。

Objective To observe the effects of Stomach-Harmonizing and Qi-Soothing Recipe on gastrointestinal motility and stem cell factor(SCF)/c-kit signaling pathway in model rats with functional dyspepsia of liver depression,and to explore its mechanism at molecular level.Methods Forty SPF healthy Wistar rats,who were selected according to approximate scores for open-field test and consumption of saccharose,were divided into normal group,model group,Chinese herb group,western drug group,10 rats in each group.Apart from the normal group,the rats in the other groups were induced into functional dyspepsia model of liver depression by clipping tail plus irregular feeding methods.After successful modeling,the rats in the Chinese herb group and western drug group were given intervention treatment with Stomach-Harmonizing and Qi-Soothing Recipe and Domperidone,respectively.After medication,whether the rats were in depressive state was evaluated by openfield test and consumption of saccharose,state of gastrointestinal motility was assessed by gastric emptying test and small intestine propulsion test,protein expression levels of c-kit and SCF in antrum and small intestinal tissue were measured by immunohistochemical assay and Western blotting assay,mRNA expression levels of c-kit and SCF in antrum and small intestinal mucosa were detected by real time quantitative polymerase chain reaction(RTPCR)assay.Results Compared with the normal group,the saccharose consumption rate was reduced in the model group,horizontal and vertical movement scores of rats in open-field test were lowered,stomach emptying and small intestine propulsion were slowed,optical density values of c-kit and SCF positively expressive cells in the antrum and small intestinal tissue were decreased,and protein and mRNA expression levels of c-kit and SCF were decreased(all P<0.05).Compared with the model group,the saccharose consumption rate was enhanced in the Chinese herb group and western drug group,horizontal and vertical movement scores of rats in open-field test were raised,stomach emptying and small intestine propulsion were quicken,optical density values of c-kit and SCF positively expressive cells in antrum and small intestinal tissue were increased,and protein and mRNA expression levels of c-kit and SCF were increased(all P<0.05),while there was no statistical differences in the above indexes between Chinese herb group and western drug group(P>0.05).Conclusion Liver depression can lead to gastrointestinal motility dysfunction,Stomach-Harmonizing and Qi-Soothing Recipe is effective for improving depression state of functional dyspepsia rats with liver depression,and its molecular mechanism is related to upregulating expression of relevant proteins in SCF/c-kit signaling pathway.