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目的探讨固本通络方治疗IgA肾病的作用机制。方法昆明种小鼠经过12周诱导建立IgA肾病动物模型,随机分为模型组(n=15)、中药组(n=15);另取正常小鼠作为正常对照组(n=15)。12周造模结束时开始,中药组按小鼠体质量予固本通络方1.35mL/(g·d)灌胃,正常对照组和模型组予等量生理盐水灌胃,每日1次,均连续8周。第0周、12周和20周时,测定各组24h尿蛋白;第12周和20周观察肾组织病理学改变,第20周时实时荧光定量PCR检测Peyer小结转化生长因子β(TGF-β)、细胞信号转导分子(Smad)3mRNA表达,免疫组化法检测Peyer小结TGF-β、Smad3蛋白的表达,流式细胞测定Peyer小结(IgA+B淋巴细胞)/B淋巴细胞水平。结果与正常对照组相比,模型组小鼠24h尿蛋白升高(12周和20周),系膜区IgA沉积,Peyer小结TGF-β、Smad3蛋白和mRNA表达升高,(IgA+B淋巴细胞)/B淋巴细胞水平升高,差异均有统计学意义(P均<0.01)。与模型组相比,中药组各项指标均有所改善,差异有统计学意义(P均<0.01);但与正常对照组相比,中药组各项指标的差异有统计学意义(P均<0.01)。结论固本通络方能有效改善IgA肾病小鼠蛋白尿及肾小球系膜区IgA沉积,可能与其调节Peyer小结TGF-β/Smad3,进而影响B淋巴细胞IgA类别转换有关。
Objective To explore the mechanism of Guben Tongluo Recipe in treating IgA nephropathy. Methods Kunming mice were induced to establish IgA nephropathy model after 12 weeks and were randomly divided into model group (n = 15) and TCM group (n = 15). Normal mice were used as normal control group (n = 15). At the end of 12 weeks, the rats in TCM group were given Guben Tonglu formula (1.35mL / (g · d)) by intragastric administration. The normal control group and model group were given the same amount of normal saline once a day, All for 8 weeks. The urinary protein of 24 hours was measured at the 0th week, the 12th week and the 20th week. The pathological changes of the kidney were observed at the 12th week and the 20th week. The expression of Peyer’s tubule transforming growth factor beta (TGF-β) was detected by real-time fluorescence quantitative PCR ), Smad 3 mRNA expression. The expression of TGF-β and Smad3 protein in Peyer’s nodule was detected by immunohistochemistry. The level of Peyer’s nodule (IgA + B lymphocyte) / B lymphocyte was determined by flow cytometry. Results Compared with the normal control group, 24h urinary protein (12 weeks and 20 weeks), mesangial IgA deposition, Peyer’s tubulointerstitium increased the expression of TGF-β, Smad3 protein and mRNA in model group (IgA + B lymph Cells) / B lymphocytes increased, the difference was statistically significant (P all <0.01). Compared with the model group, all the indexes in the traditional Chinese medicine group improved significantly (P <0.01), but there were significant differences among the indexes of the traditional Chinese medicine group (P <0.01) <0.01). Conclusion Guben Tongluo Recipe can effectively improve the proteinuria and IgA deposition in IgA nephropathy mice, which may be related to the regulation of Peyer’s tubulointerstitial TGF-β / Smad3 and IgA type conversion in B lymphocytes.