益气活血药对心肌梗死后大鼠小肠黏膜屏障及肠道菌群的影响

Effects of Qi-Boosting and Blood-Moving Decoction on small intestinal mucosal barrier and intestinal flora of rats after myocardial infarction

目的观察益气活血药对心梗后大鼠肠道黏膜屏障、肠道菌群及封闭蛋白的影响。方法采用结扎左冠状动脉前降支制备 SD 大鼠的心肌梗死模型,将造模成功的大鼠随机分为模型组、益气活血药组、酒石酸美托洛尔组,连同假手术组,每组 10 只。实验分为 7 天和 28 天两个时间节点。术后第 2 天开始灌胃给药,益气活血方组 8. 2 g/( kg·d)、酒石酸美托洛尔组 5 mg/( kg·d),模型组和假手术组给予等量蒸馏水灌胃。分别于灌胃 7 天和 28 天后采集大鼠小肠(回肠)组织及食糜。HE 染色观察小肠(回肠)黏膜结构,测量绒毛高度、隐窝深度,并计算绒毛高度/隐窝深度( Villus height / Crypt depth ,V/C)比值;实时荧光定量 PCR方法观察小肠有益菌(双歧杆菌、乳杆菌)和有害菌(大肠杆菌、和肠球菌)的变化;免疫组化方法检测封闭蛋白表达的变化。结果在 7 天和 28 天两个时间节点中,与假手术相比,模型组绒毛高度下降( P < 0. 01),隐窝深度变浅( P <0. 01),V/C 比值减小( P < 0. 01),Occludin 蛋白表达明显减少( P < 0. 01),有益菌数量均明显减少( P <0. 01),有害菌数量明显增加( P < 0. 01);与模型组相比,各给药组绒毛高度明显升高( P <0. 01),隐窝变浅( P <0. 05),V/C 比值明显增大( P <0. 01),益气活血药组有益菌数量显著增加( P <0. 01),有害菌数量显著减少( P <0. 01),两用药组趋势相同,两者数量无统计差异,益气活血药组 Occludin 蛋白的表达增加( P <0. 01)。结论本实验发现心梗大鼠小肠形态及功能受到影响,益气活血药干预可改善心梗后大鼠肠道形态结构,提高小肠的消化与吸收能力,调节小肠菌群。

Objective To observe the effects of Qi-Boosting and Blood-Moving ( QBBM) Decoction on intestinal mucosal barrier,intestinal flora and Occludin protein in rats after myocardial infarction. Meth- ods The myocardial infarction model of SD rats was established by ligation of left anterior descending coro- nary artery. Rats with successful modeling were randomly divided into model group,QBBM Decoction group, metoprolol tartrate group,and sham operation group,each group has 10 rats. There were 2 time nodes during the experiment: 7d and 28d. On the 2 nd day after the operation,the rats were given intragastric administra- tion: the QBBM Decoction group 8. 2g/( kg·d),the metoprolol tartrate group 5mg/( kg·d);the model group and the sham group were given equal amount of distilled water to the stomach. Small intestinal ( ileal) tissue and chyme of the rats were collected after 7 and 28 days of gavage,respectively. HE staining was used to observe the structure of small intestinal ( ileal) mucosa,Villus height and Crypt depth were measured,and the V/C ratio was calculated. RT-PCR was used to observe the changes of beneficial bacteria ( bifidobacteria and lactobacilli) and harmful bacteria ( Escherichia coli and enterococci) of the small intestine;the immunohistochemical method was used to detect the changes in Occludin protein expression. Results On the 7d and 28d time nodes,compared with the sham operation group,the Villus height of the model group decreased ( P <0.01),the Crypt depth became shallower ( P <0.01),the V/C ratio decreased ( P <0. 01),Occludin protein expression decreased significantly ( P <0.01),the number of beneficial bacteria were significantly decreased ( P <0. 01),and the number of harmful bacteria increased significantly ( P <0. 01);Compared with the model group,the Villus height was obviously increased ( P <0.01),the Crypt depth became shallo- wer ( P <0.05),and the V/C ratio increased obviously ( P <0.01) in each groups. The beneficial bacteria number of the QBBM Decoction group was increased significantly ( P <0.01),the number of harmful bacte- ria was significantly reduced ( P <0.01),the trend of each medicinal group was the same,the number of which had no statistical difference,and the Occludin protein expression of the QBBM Decoction group was increased ( P <0.01). Conclusion In this study,it was found that the morphology and function of small intestine of the rats with myocardial infarction were affected. The intervention of QBBM Decoction could improve the intestinal morphology and structure of rats with myocardial infarction,improve the ability of digestion and absorption of the small intestine,and regulate the small intestinal flora.