摘要
目的通过观察keap-1-Nrf2通路调控claudin-2蛋白表达变化,探讨健脾清热活血方防治溃疡性结肠炎的效用机制。方法90只SPF级雄性SD大鼠随机分为6组,除外正常组,其余各组均采用TNBS灌肠法制备溃疡性结肠炎模型,自第7日起分组给药,模型组予等剂量生理盐水,中药低、中、高剂量组予不同剂量健脾清热活血方,西药组予美沙拉嗪灌胃,连续14d。末次给药后腹腔麻醉处死全部大鼠,截取病变部位肠段备测。HE染色检测结肠组织病理学变化;透射电镜检测结肠黏膜超微结构变化;免疫组化SP法检测结肠环氧氯丙烷结合蛋白1(keap-1)、Nrf2以及claudin-2蛋白表达变化。结果光镜提示模型组结肠黏膜表面出现缺损或脱落坏死,溃疡形成,隐窝结构消失、脓肿形成,固有层大量炎性细胞浸润,黏膜下层充血肿胀;治疗组较模型组有明显改善,见结肠黏膜尚完整,但存在黏膜缺损,隐窝脓肿,部分可见出血点,较多炎性细胞浸润固有层。电镜下见模型组结肠上皮细胞微绒毛稀疏,线粒体肿胀变大,细胞间紧密连接消失,可见细胞处于坏死期;中药组见上皮细胞体积明显变小、核固缩、凋亡小体形成,局灶可见坏死。模型组结肠keap-1、Nrf2蛋白表达下降,claudin-2蛋白表达上升;治疗后,中药组结肠keap-1、Nrf2蛋白表达上升,claudin-2蛋白表达下降;经比较,差异有统计学意义(P<0.05);但是中药组之间不存在组间差异(P>0.05)。结论健脾清热活血方通过上调Keap-1、Nrf2表达,下调claudin-2表达而发挥防治溃疡性结肠炎的效用。
Objective:To observe the effect of keap-l-Nrf2 pathway on the expression of claudin-2 protein,and to explore the mechanism of Jianpi Qingre Huoxue Decoction in preventing and treating ulcerative colitis.Methods:90 SPF male SD rats were randomly divided into 6 groups according to body weight,except for the normal group.The rest of the groups were prepared by TNBS enema method for ulcerative colitis models.They were administered in groups starting from the 7 th day.The saline and treatment groups were given Jianpi Qingre Huoxue Decoction with different doses,and the control group was given gavage by mesalazine for 14 days.All rats were sacrificed by intraperitoneal anesthesia after the last administration,and the intestinal segment of the lesion was taken for testing.HE staining was used to detect the pathological changes of colonic tissue;transmission electron microscopy was used to detect the ultrastructural changes of colonic mucosa;immunohistochemical SP method was used to detect the expression of colon keap-1,Nrf2 and claudin-2.Results:Light microscopy showed that the surface of the colonic mucosa in the model group had defects or shedding and necrosis,ulcers formed,crypt structures disappeared,abscesses formed,a large number of inflammatory cells infiltrated the lamina propria,and the submucosa was congested and swollen.The treatment group had a more significant improvement than the model group.Under the light microscopy,the colonic mucosa was still intact,but there were mucosal defects,crypt abscesses,some bleeding points,and more inflammatory cells infiltrated the lamina propria.The electron microscopy showed that the microvilli of the colonic epithelial cells in the model group were sparse,the mitochondria became swollen and enlarged,and the tight junctions between the cells disappeared.The cells were in the necrotic phase.The treatment group had epithelial cells significantly smaller in size,the pyknosis,and the formation of apoptotic bodies.Necrosis was visible on the stove.The expression of keap-1 and Nrf2 protein in the model group decreased,and the expression of claudin-2 protein increased.After treatment,the expression of keap-1 and Nrf2 protein in the treatment group increased,and the expression of claudin-2 protein decreased.After comparison,there was a statistical difference(P<0.05);however,there were no differences between the treatment groups(P>0.05).Conclusion:Jianpi Qingre Huoxue Decoction exerts the effect of preventing and treating ulcerative colitis by upregulating the expression of keap-1 and Nrf2 and down-regulating the expression of claudin-2.
作者
罗淑娟
陈小芬
林逸婷
卢鑫
张涛
Luo Shujuan;Chen Xiaofen;Lin Yiting;Lu Xin;Zhang Tao(Faculty of Chinese Medicine Science Guangxi University of Chinese Medicine,Guongxi,Nanning 530222,China)
出处
《中国中医急症》
2021年第11期1924-1928,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
广西自然科学基金项目(2018GXNSFAA281042)。
作者简介
通信作者:张涛,电子邮箱:327664246@qq.com。
