葛根芩连汤加减对糖尿病大鼠海马炎症因子的影响

Effect of Modified Gegen Qinlian Tang on Inflammatory Factors in Hippocampus for Diabetic Rats

目的:探讨葛根芩连汤加减对2型糖尿病(T2DM)大鼠认知功能及海马组织白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)和核转录因子-κB(NF-κB)蛋白表达的影响。方法:将62只大鼠随机选取12只作为空白组,其余50只以高脂高糖饮食及小剂量链脲佐菌素(STZ)腹腔注射诱导2型糖尿病模型,随机分为模型组、沙格列汀组、葛根芩连汤加减组。空白组和模型组大鼠分别灌胃蒸馏水10 mL·kg-1·d-1,沙格列汀组灌胃沙格列汀0.005 g·kg-1·d-1,葛根芩连汤加减组灌胃葛根芩连汤加减0.435 g·kg-1·d-1;8周后,采用Morris水迷宫观察大鼠学习记忆能力;苏木素-伊红(HE)染色法观察海马组织病理变化;蛋白免疫印迹法(Western blot)检测海马组织细胞浆和细胞核内NF-κB p65蛋白的表达;酶联免疫吸附测定(ELISA)检测大鼠海马组织IL-1β,TNF-α的水平。结果:与正常组比较,模型组大鼠第4天平均潜伏期显著延长(P<0.01),逃逸平台进入次数、有效区域运动时间和运动距离均显著降低(P<0.01),细胞核内NF-κB p65的蛋白表达显著上升(P<0.01),大鼠海马中IL-1β,TNF-α表达水平明显升高(P<0.05);与模型组比较,葛根芩连汤加减能明显缩短T2DM大鼠逃避潜伏期,增加穿越平台次数、有效区游泳路程及游泳时间(P<0.05,P<0.01),显著改善海马组织IL-1β,TNF-α和细胞核内NF-κB p65蛋白表达水平(P<0.05,P<0.01)。结论:葛根芩连汤加减可以显著改善高糖高脂饮食加小剂量STZ诱导的T2DM大鼠模型的学习记忆能力,其作用机制可能是通过抑制大鼠海马炎症反应来实现的。

Objective:To investigate the effect of modified Gegen Qinlian Tang on learning and memory ability and interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),nuclear factor-κB(NF-κB)protein expression in hippocampus of type 2 diabetic(T2DM)rats.Method:A total of 62 rats were randomly divided into normal group(n=12),and the remaining 50 rats were established by feeding high sugar and high fat diet and injecting with low dosage of streptozocin(STZ)to T2DM.The successful modeling rats were selected and randomly divided into model group,Saxagliptin and modified Gegen Qinlian Tang group.The normal group and the model group received 10 mL·kg-1·d-1 nomal saline by intragastric administration.0.005 g·kg-1·d-1 Saxagliptin crude drug and 0.435 g·kg-1·d-1 modified Gegen Qinlian Tang crude drug were given in Saxagliptin group and modified Gegen Qinlian Tang group,and all the treatment lasted for 8 weeks.Learning and memory ability of rats was observed by using Morris water maze.Histopathological changes of hippocampus was observed by hematoxylineosin staining.Western blot was used to test the NF-κB p65 in the cytoplasmic and nucleus in rat hippocampus.Enzyme-linked immunosorbent assay(ELISA)was used to test the levels of IL-1βand TNF-α.Result:Compared with normal group,the mean incubation period of model group was significantly prolonged on the 4 thday(P<0.01),the number of escape platform entry,the movement time and distance of the effective area were significantly decreased(P<0.01),the protein expression of NF-κB p65 in the nucleus was significantly increased(P<0.01),and the expression levels of IL-1β,TNF-αin the hippocampus were significantly increased(P<0.05).Compared with normal group,modified Gegen Qinlian Tang could increase the number of crossing platform(P<0.05,P<0.01),the platform of swimming distance and swimming time(P<0.05,P<0.01).The expression levels of IL-1β,TNF-αand NF-κB p65 in hippocampus were significantly improved.Conclusion:Modified Gegen Qinlian Tang could obviously improve the learning and memory ability of the T2DM rat model which was induced by the high-sugar,high-lipid diet and low-dose STZ,and the mechanism may be achieved by suppressing the inflammation of the rat’s hippocampus.