上海交通大学学报(医学版) ›› 2018, Vol. 38 ›› Issue (6): 647-.doi: 10.3969/j.issn.1674-8115.2018.06.011

• 论著·临床研究 • 上一篇    下一篇

不同肾功能进展IgA肾病患者血清IgA1对人肾小球系膜细胞增殖及TGF-β1表达的影响

孔凌云,蒋更如   

  1. 上海交通大学医学院附属新华医院肾内科,上海 200092
  • 出版日期:2018-06-28 发布日期:2018-07-03
  • 通讯作者: 蒋更如,电子信箱:jianggeng-ru@hotmail.com。
  • 作者简介:孔凌云(1992—),女,硕士生;电子信箱:18800204302@163.com。
  • 基金资助:
    上海市科学技术委员会基金(12140903002);上海市卫生和计划生育委员会科研课题(20144Y0145)

Effect of serum IgA1 on human mesangial cells and of TGF-β1 in patients of IgA nephropathy with different renal function progress

KONG Ling-yun, JIANG Geng-ru   

  1. Department of Nephrology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2018-06-28 Published:2018-07-03
  • Supported by:
    Science and Technology Commission of Shanghai Municipality, 12140903002; Projects of Shanghai Municipal Health and Family Planning Commission, 20144Y0145

摘要: 目的·探讨肾功能快速进展为终末期肾病(end stage renal disease,ESRD)的IgA肾病患者与肾功能长期稳定的IgA肾病患者血清IgA1对体外培养的正常人肾小球系膜细胞(human mesangial cells,HMCs)增殖的刺激作用及对转化生长因子β1(transforming growth factor-β1,TGF-β1)表达水平的影响。方法· 9例经肾活检确诊为原发性IgA肾病的患者,根据病情进展情况分为肾功能快速进展组(n6)和肾功能长期稳定组(n3)。采用亲和层析及分子筛提取血清IgA1;用不同质量浓度(10、50、250、1 000 μg/mL)的IgA1分别干预HMC 12、24 h。CCK8法测定细胞增殖率;real-time PCR测定TGF-β1 mRNA表达;ELISA法测定TGF-β1蛋白表达。结果·肾功能快速进展组及肾功能长期稳定组患者血清IgA1对HMC有明显的促增殖作用,且表现为时间依赖性和浓度依赖性,作用高峰质量浓度分别为250 μg/mL和1 000 μg/mL。2组患者血清IgA1均能显著上调TGF-β1的mRNA和蛋白的表达。在肾功能快速进展组,IgA1质量浓度为10 μg/mL时TGF-β1的mRNA及蛋白水平达到高峰,其后随刺激浓度升高而递减;在肾功能长期稳定组,TGF-β1水平则与血清IgA1呈现平行的浓度依赖性。结论·肾功能快速进展及长期稳定的患者血清IgA1在一定浓度和时间范围内均具有刺激HMC增殖的作用。在较低质量浓度(10~50 μg/mL)范围内,促增殖作用没有明显差异,但肾功能快速进展的患者血清IgA1上调TGF-β1表达的作用更强,提示血清IgA1具有一定的预后评估作用。

关键词: IgA肾病, 肾功能, 系膜细胞, 转化生长因子&, beta, 1

Abstract:

Objective · To extract serum IgA1 patients with IgA nephropathy (IgAN) (end stage renal disease vs long-term srenal function) to explore the effect on proliferation rate of human mesangial cells (HMCs) and the level of transforming growth factor-β1 (TGF-β1). Methods · Nine patients with primary IgAN were divided into rapidly progressive group (n6) and long-term sgroup (n3). Jacalin affinity chromatography and sephacryl S-200HRmolecular sieves were used to distract serum IgA1. HMCs were cultured and co-cultivated with different IgA1 concentration (10, 50, 250 and 1 000 μg/mL)at point of 12 h and 24 h respectively. The proliferation rate was measuredcell counting kit-8 (CCK8). The of TGF-β1 mRNA was measured via quantitative real-time PCR (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to detect TGF-β1 protein. Results · Serum IgA1 IgAN patients with different renal functions (end stage renal disease vs long-term srenal function) activated proliferation of HMC significantly, presenting with time-dependence and concentration-dependence. The highest value showed at 250 μg/mL and 1 000 μg/mL respectively. Serum IgA1 in two groups of patients statistically increased the of TGF-β1 mRNA and protein. In group with end stage renal disease, the summit stood at 10 μg/mL and started to decreasedegrees afterwards; while in group with long-term srenal function, the level of TGF-β1 increased in a parallel manner with the serum IgA1. Conclusion · Serum IgA1 IgAN patients with different renal functions (end stage renal disease vs long-term srenal function) can both promote the proliferation of HMC. There is no dramatical difference observed with in10-50 μg/mL, but the IgA1 group with end stage renal disease reveals a stronger effect on TGF-β1, in accordance with the pathological type of the patients (IgA sclerosis), suggesting the prognostic value of serum IgA.

Key words: IgA nephropathy, renal function, human mesangial cell, transforming growth factor-&, beta, 1

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