厚荷叶和乌檀叶对肠易激综合征大鼠TRPA1和TRPM8表达的影响

Jiahui HE , Yuying ZHANG , Chaoyan YANG , Kunping LI , Xujiang YUAN , Zihong ZHANG , Chiming GU , Yanfen CHEN
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引用次数: 0

摘要

目的厚朴和乌龙果是两种具有温性的中草药,通常被称为温内药,历史悠久,广泛用于治疗感冒证。本研究旨在探讨枳实、枳实对肠易激综合征(IBS)脾胃虚寒证大鼠的影响,并阐明“温中止痛”与瞬时受体电位通道A1 (TRPA1)、M8 (TRPM8)之间的关系。方法采用寒泻法和抑制应激法建立脾胃虚寒证大鼠模型。将大鼠随机分为正常组、模型组、阳性组(溴化匹维铵)、铁皮石斛组和金针叶组。连续造模给药7天后,测定大鼠体重、稀便等级、ATP酶活性、胃肠道组织病理形态。采用RT-PCR和免疫组织化学方法检测大鼠脊髓神经节、胃和结肠组织中TRPA1和TRPM8的表达。结果脾胃虚寒证型肠易激综合征大鼠腹泻次数增加,体重减轻。与正常组比较,模型组大鼠稀便等级显著升高(P<0.01),红细胞膜Na+-K+-ATP和Ca2+-Mg2+-ATP活性显著降低(P<0.01)。但与模型组相比,高陵草和龙脑草明显降低了大鼠稀便等级(P<0.05或P<0.01),明显提高了Na+-K+-ATP和Ca2+-Mg2+-ATP的活性(P<0.05)。模型组大鼠脊髓神经节和结肠组织中TRPA1和TRPM8的表达明显升高(P<0.01),而高良、芦茱萸显著抑制了TRPA1和TRPM8的过表达。结论枳实、乌龙果对IBS模型的温热镇痛作用可能主要通过调节TRPA1、TRPM8的异常表达来实现。因此,这些结果可以支持温性中药如高良、乌龙果等传统上用于治疗脾胃虚寒证的事实。
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Effects of Alpinia Officinarum and Euodia Rutaecarpa on the expression of TRPA1 and TRPM8 in rats with irritable bowel syndrome

Objective

Alpinia Officinarum and Euodia Rutaecarpap are two traditional Chinese herbs with warm nature, usually called interior-warming medicinal and widely used to treat cold syndrome for a long history. The purpose of this study is to explore the effects of Alpinia Officinarum and Euodia Rutaecarpap on irritable bowel syndrome (IBS) rats with spleen-stomach deficiency cold syndrome, and to clarify the link between “warming the middle to alleviate pain” and transient receptor potential channel A1 (TRPA1), M8 (TRPM8).

Methods

IBS rat model with spleen-stomach deficiency cold syndrome was established by cold diarrhea and restraint stress. Rats were randomly divided into the normal group, model group, positive group (pinaverium bromide), Alpinia Officinarum group and Euodia Rutaecarpa group. After continuous modeling and administration for seven days, body weight, loose stool grades, activity of ATP enzyme, pathological morphology of gastrointestinal tissue were evaluated. The expression of TRPA1 and TRPM8 in spinal ganglion, stomach and colon tissue of rats were detected by RT-PCR and immunohistochemistry.

Results

The IBS model with spleen-stomach deficiency cold syndrome caused increase in the number of diarrhea and loss in the body weight in rats. Compared with the normal group, the loose stool grades of model group was significantly increased (P<0.01), Na+-K+-ATP and Ca2+-Mg2+-ATP activity of erythrocyte membrane were significantly decreased (P<0.01). However, compared with the model group, Alpinia Officinarum and Euodia Rutaecarpa obviously reduced the loose stool grades (P<0.05 or P<0.01), and obviously increased the activity of Na+-K+-ATP and Ca2+-Mg2+-ATP (P<0.05). The expression of TRPA1 and TRPM8 in spinal ganglia and colon tissue of the model group was markedly enhanced (P<0.01), but Alpinia Officinarum and Euodia Rutaecarpa significantly inhibited these overexpression.

Conclusion

The effect of interior-warming and alleviating pain of Alpinia Officinarum and Euodia Rutaecarpa on IBS model may be mainly achieved by regulating the abnormal expression of TRPA1 and TRPM8. Thus, these results could support the fact that interior-warming medicinal such as Alpinia Officinarum and Euodia Rutaecarpa are traditionally used to cure spleen-stomach deficiency cold syndrome.

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