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目的:观察SIRT1在慢传输型便秘中的表达情况,并探讨通便汤恢复慢传输型便秘大鼠结肠动力的作用机制。方法:采用大黄酸法制备大鼠慢传输型便秘模型,造模组40只大鼠,大鼠随机分为模型组、通便汤低、中和高剂量组(每组各10只),另取10只作为正常组。记录大鼠一般情况、粪便含水量、测定结肠肌电变化、肠道传输功能及结肠超微结构情况,并运用免疫组织化学技术检测结肠SIRT1蛋白的分布、表达及相对含量。利用RT-PCR法测SIRT1、FoxO1mRNA表达情况并进行统计学分析。结果:与模型组相比,通便汤高剂量组可显著增加粪便含水量(P<0.05),通便汤中、高剂量组可降低频率和振幅变异系数(P<0.05),稳定结肠慢波节律。除此之外,通便汤低、中和高剂量组可显著减少肠嗜铬细胞和神经分泌颗粒。免疫组化结果显示通便汤低、中、高剂量组能不同程度提高SIRT1蛋白表达(P<0.05),以通便汤高剂量组最为明显。RT-PCR结果显示通便汤各剂量组大鼠结肠组织SIRT1、FoxO1mRNA显著升高(P<0.05)。结论:慢传输型便秘发生发展过程中SIRT1信号通路发生改变,通便汤恢复结肠动力的机制可能通过提高SIRT1活性,使得FoxO1去乙酰化,增强FoxO1活性。
Objective: To observe the expression of SIRT1 in slow transit constipation and to explore the mechanism of Tongbang decoction in relieving colonic motility in slow transit constipation rats. Methods: The model of slow transit constipation was established by rhein method. Forty rats were randomly divided into model group, low, medium and high dose group (10 rats in each group) Take 10 as a normal group. The general situation of rat, the water content of feces, the changes of EMG, the intestinal transit function and the ultrastructure of colon were recorded. The distribution, expression and relative content of SIRT1 protein in colon were detected by immunohistochemistry. The expression of SIRT1 and FoxO1 mRNA was detected by RT-PCR and analyzed statistically. Results: Compared with the model group, the high dose group of Tongbang decoction could significantly increase the water content of feces (P <0.05), and the medium and high dose of Tongbang decoction could reduce the coefficient of variation of frequency and amplitude (P <0.05) Wave rhythm. In addition, the low, medium and high dose group of STT can significantly reduce enterochromaffin cells and neurosecretory granules. The results of immunohistochemistry showed that the low, medium and high dose of Tongmai Decoction could increase the expression of SIRT1 protein to some extent (P <0.05), and the most obvious was Tongmai Decoction high dose group. The results of RT-PCR showed that the mRNA levels of SIRT1 and FoxO1 in colonic tissue of rats in each group were significantly increased (P <0.05). CONCLUSIONS: The SIRT1 signaling pathway changes during the development of slow transit constipation. The mechanism of Tongmao soup restoring colonic motility may deacetylate FoxO1 and enhance FoxO1 activity by increasing SIRT1 activity.