The protective effect of taurochenodeoxycholic acid on experimental enteritis injury in rabbits:a preliminary study
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摘要:
目的:分析牛磺鹅去氧胆酸(taurochenodeoxycholic acid,TCDCA)对肠道炎症的疗效,为肠道急慢性炎症的治疗提供理论依据。方法:体内外实验分组均为对照组、脂多糖(lipopolysaccharide,LPS)组、LPS+TCDCA组。体外实验中首先应用MTT 法筛选TCDCA的适宜工作浓度,随后LPS组给予巨噬细胞LPS刺激,LPS+TCDCA组先后给予LPS与TCDCA刺激,RT-qPCR和 Western blot检测炎症相关mRNA和蛋白的表达。体内实验中,LPS组通过兔耳缘静脉注射LPS溶液,LPS+TCDCA组同上处理后通过饮水喂食TCDCA溶液。通过HE染色、过碘酸-雪夫染色和阿尔新蓝-核固红染色评估小肠的组织病理学改变。结果:体外实验结果显示,与LPS组相比,LPS+TCDCA组巨噬细胞中肿瘤坏死因子-α、白介素-1β、白介素-6、干扰素-γ表达量显著降低,白介素-10表达量升高。体内实验中,HE染色结果显示小肠组织炎症缓解,过碘酸-雪夫染色和阿尔新蓝-核固红染色结果显示LPS+TCDCA组杯状细胞数量和酸性、中性黏蛋白分泌量均较LPS组增多。结论:TCDCA可通过降低炎症因子的表达缓解兔肠道组织炎症,减轻LPS造成的肠道炎症损伤。
Abstract:
Objective:To analyze the efficacy of taurochenodeoxycholic acid(TCDCA)on intestinal inflammation and to provide a theoretical basis for the treatment of acute and chronic intestinal inflammation. Methods:In the in vitro and in vitro experiments,the cells and animals were devided into the control group,lipopolysaccharide(LPS)group,and LPS + TCDCA group. In the in vitro experiments,MTT method was firstly applied to screen the appropriate working concentration of TCDCA,and then LPS stimulation was given to the LPS group,and LPS and TCDCA stimulation were given to the LPS + TCDCA group subsequently. The expression of inflammation-related mRNAs and proteins were examined by RT-qPCR and Western blot,respectively . In the in vivo experiments,the LPS group was injected with LPS solution intravenously through the ear margin of rabbits,and the LPS+TCDCA group was treated as above and then fed with TCDCA solution through drinking water. Histopathological alterations of the samples were assessed by HE staining,periodic acid-Schiff staining and Alcian blue-nuclear fast red staining after experimental sampling. Results:The results of in vitro experiments showed that the expression of tumor necrosis factor-α,interleukin-1β,interleukin-6,and interferon-γ indexes was significantly reduced and the expression of interleukin-10 indexes was elevated in macrophages in the LPS+TCDCA group compared with that in the LPS group. In the in vivo experiments,the results of HE staining showed that the inflammation of intestinal tissues was relieved,and the results of periodic acid-Schiff staining and Alcian blue-nucleic solid red staining showed that the number of cup cells and the secretion of acidic as well as neutral mucins in the LPS + TCDCA group increased compared with those in the LPS group. Conclusion:TCDCA alleviates intestinal tissue inflammation and reduces intestinal inflammatory damage caused by LPS by decreasing the expression of inflammatory factors.