目的探讨黄芪甲苷(AS-IV)对IgA肾病(IgAN)小鼠辅助性T细胞1(Th1)和Th2细胞平衡的影响及其可能机制。方法建立BALB/c小鼠IgAN模型。模型成功的小鼠随机分为四组:模型,AS-IV低剂量,AS-IV中等剂量组和AS-IV高剂量组,每组10只小鼠。另外10只小鼠作为对照组。老鼠在低处,中、高剂量组分别灌胃12.5、25、50mg/kgAS-IV混悬液(生理盐水配制),对照组和模型组给予等量生理盐水。测定各组24小时尿蛋白(24hUPr)含量和尿红细胞计数。血尿素氮(BUN)的水平,测定血清肌酐(Scr)和白蛋白(ALB)。血清干扰素γ(IFN-γ),ELISA法检测白细胞介素4(IL-4)和IL-10水平。应用流式细胞术检测小鼠外周血中Th1/Th2细胞的比例。HE染色观察小鼠肾脏的组织病理学变化。RT-PCR和Westernblot检测T细胞免疫球蛋白和粘蛋白结构域基因1(TIM-1)的mRNA和蛋白表达,小鼠肾组织中的Toll样受体4(TLR4)。结果与模型组比较,在12周和15周,尿液红细胞计数,24hUPr,BUN,Scr,IL-4和IL-10的水平,Th2细胞的比例,TIM-1和TLR4的mRNA和蛋白表达水平显著降低,中、高剂量组AS-IV,以及ALB的水平,IFN-γ,Th1细胞比例和Th1/Th2细胞比例增加,高剂量组效果最好。结论AS-IV可抑制TIM-1信号通路,增加Th1/Th2细胞比例,抑制炎症反应,减轻IgAN小鼠肾损伤。
Objective To investigate the effects of astragaloside IV(AS-IV) on the balance of T helper type 1 (Th1) and Th2 cells in mice with IgA nephropathy (IgAN) and its possible mechanism. Methods The IgAN model of BALB/c mice was established. Successfully modeled mice were randomly divided into four groups: model, AS-IV low dose, AS-IV medium dose and AS-IV high dose groups, with 10 mice in each group. Another 10 mice served as the control group. Mice in the low, medium and high dose groups were administered 12.5, 25 and 50 mg/kg AS-IV suspension (prepared in normal saline) by gavage, while the control and model groups were given an equivalent volume of normal saline. The 24-hour urinary protein (24 h UPr) content and urine red blood cell count were measured in each group. The levels of blood urea nitrogen (BUN), serum creatinine (Scr) and albumin (ALB) were determined. Serum interferon γ (IFN-γ), interleukin 4 (IL-4) and IL-10 levels were detected by ELISA. The ratio of Th1/Th2 cells in peripheral blood of mice was detected using flow cytometry. Histopathological changes in the kidney of mice were observed by HE staining. RT-PCR and Western blot were used to detect the mRNA and protein expressions of T cell immunoglobulin and mucin domain gene 1 (TIM-1), Toll-like receptor 4 (TLR4) in mouse kidney tissue. Results Compared with the model group, in weeks 12 and 15, the urine red blood cell count, 24 h UPr, BUN, Scr, levels of IL-4 and IL-10, the proportion of Th2 cells, as well as the mRNA and protein expression levels of TIM-1 and TLR4 were significantly decreased in the low, medium and high dose groups of AS-IV, and the levels of ALB, IFN-γ, the proportion of Th1 cells and Th1/Th2 cell ratio were increased, with the high-dose group showing the best effects. Conclusion AS-IV can inhibit TIM-1 signaling pathway, increase the Th1/Th2 cell ratio, inhibit the inflammatory reaction, and alleviate the renal injury in IgAN mice.