痞痛舒降低内脏高敏感治疗功能性消化不良的研究

The Mechanism of Pitongshu in Treating Functional Dyspepsia

[目的]研究痞痛舒对功能性消化不良(functional dyspepsia,FD)大鼠胃黏膜肥大细胞(mast cells,MC)活化、脊髓背角c-fos基因和蛋白表达的影响,以及对细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)磷酸化水平的影响。[方法]100只SD大鼠随机分为正常对照组、模型组、痞痛舒低剂量组、痞痛舒中剂量组、痞痛舒高剂量组,以碘乙酰胺蔗糖法制作FD大鼠模型,造模成功后给药28d,各组大鼠行胃肌电图(electromyography,EMG)检查,甲苯胺蓝染色法观察胃黏膜MC并计数,免疫组化检测脊髓背角c-fos蛋白表达、Real-time RT-PCR检测c-fos m RNA表达、Western blot检测ERK1/2磷酸化水平。[结果]1.模型组胃EMG振幅、胃组织黏膜MC数、脊髓背角c-fos蛋白、c-fos m RNA表达明显高于正常对照组(P<0.05),HE染色镜下观察胃组织未见明显炎症改变。2.与模型组相比,痞痛舒高、中、低剂量组大鼠胃组织MC数量明显减少(P<0.05),但各组间差异无统计学意义(P>0.05)。痞痛舒高剂量组的大鼠脊髓背角c-fos蛋白及m RNA表达低于模型组(P<0.05),痞痛舒中、高剂量组c-fos m RNA的表达、ERK1/2磷酸化水平低于模型组和痞痛舒低剂量组(P<0.05)。[结论]痞痛舒治疗FD的机制可能是抑制胃黏膜MC活化,减少末梢神经刺激,抑制c-fos表达和ERK磷酸化,降低中枢敏感,从而降低内脏高敏感。

[Objective] To study the effect of Pitongs.hu on gastric mncosal mast cells, c-fos and c-fos mRNA expression and the activity of ERK1/2 in spinal dorsal horn in rats with functional dyspepsia. [Methods]One hundred 10-day-old Sprague-Dawley（SD） rats were randomly divided into 5 groups： normal control group, model group and Pitongshu low, medium and high dose groups. The FD rat model was established with iodoacetamide sucrose. After administering medicine for 28 days, the rats in each group were examined by gastric electromyography. The gastric mucosal mast cells were observed by toluidine blue staining. The expression of c-los protein in spinal dorsal horn was detected by immunohistochemistry, and the expression of c-fos mRNA was detected by Real-time RT-PCR. Western blot was used to detect the ERK1/2 phosphorylation level. [Results] （1）The EMG amplitude, number of mast cells and c-fos protein, c-los mRNA expression of the model group were significantly higher than that of normal control group（P〈0.05）. And there was no significant inflammatory changes of gastric mucosa. （2）The number of MC was significantly decreased in Pitongshu medium and high dose group（P〈0.05）, but there was no statistically significant difference between the groups（P〉0.05）. The expression of c-los in high dose group was significantly lower than that of model group（P〈0.05）, medium and high dose groups of Pitongshu c-fos mRNA expression and ERK phosphorylation level were lower than that of model group and low dose groups（P〈0.05）. [Conclusion] The possible mechanism of Pitongshu was reducing mucosal mast cell numbers to reduce the stimulus on peripheral nerve and inhibiting the expression of c-fos and the activity of ERK to depress central sensitivity, thereby depressing visceral hypersensitivity.