上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (10): 1221-.doi: 10.3969/j.issn.1674-8115.2010.10.009

• 论著（临床研究） • 上一篇    下一篇

终末期肾病患者氧化应激与辅助性T淋巴细胞的关联研究

方均燕, 张 薇   

  1. 上海交通大学 医学院附属第九人民医院肾脏内科, 上海 200011
  • 出版日期:2010-10-25 发布日期:2010-10-27
  • 通讯作者: 张 薇, 电子信箱: dongzhang.1087@yahoo.com.cn。
  • 作者简介:方均燕（1982—）, 女, 硕士生；电子信箱: yankai828@yahoo.cn。
  • 基金资助:

    上海市教委基金（03BK27）

Correlation between oxidative stress and helper T lymphocytes in patients with end-stage renal disease

FANG Jun-yan, ZHANG Wei   

  1. Department of Nephrology, The Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2010-10-25 Published:2010-10-27
  • Supported by:

    Foundation of Shanghai Municipal Education Committee, China, 03BK27

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摘要：

目的 观察终末期肾病（ESRD）患者外周血辅助性T淋巴细胞和氧化应激指标的变化，探讨患者免疫功能与其氧化应激状态的关系。方法 40例ESRD患者分为血液透析治疗组（HD组）和非血液透析治疗组（NHD组），每组20例；以同期接受体检的20名健康志愿者作为正常对照组（NC组）。采集各组静脉全血，运用流式细胞仪进行T淋巴细胞计数及亚群分选，计算外周血T淋巴细胞（CD3+T细胞）、辅助性T淋巴细胞（CD4+ T细胞）百分比及其与其他T细胞亚群（CD8+T细胞）的比值（CD4+/CD8+）；检测CD4+T细胞的增殖率和凋亡率。ELISA法检测细胞培养液中细胞因子γ干扰素（IFNγ）和白介素4（IL-4）含量；黄嘌呤氧化酶法和硫代巴比妥酸分光光度法分别测定氧化应激指标血清超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平。分析各项指标间的相关性。结果 与NC组比较，HD组和NHD组的CD3+、CD4+ T细胞百分比及CD4+/CD8+比值均显著降低；CD4+ T细胞凋亡率上升，增殖率下降；IL-4含量增加，IFN-γ含量减少；血清SOD活性降低，MDA水平升高；且差异均有统计学意义（P<0.05或P<0.01）。HD组与NHD组CD3+、CD4+ T细胞百分比、CD4+ T细胞凋亡率和IL-4、IFN-γ含量比较，差异有统计学意义（P<0.05）。相关性分析表明，在ESRD患者中，外周血CD4+T细胞百分比与其细胞凋亡率呈显著负相关（r=-0.87，P＜0.01），与细胞增殖率呈显著正相关（r=0.73，P＜0.01）；CD4+T细胞凋亡率与血清MDA水平呈显著正相关（r=0.58，P＜0.01），与血清SOD活性呈显著负相关（r=-0.51，P＜0.01）；细胞培养上清液中IL-4含量与血清MDA水平呈显著正相关（r=0.47，P＜0.01）。结论 ESRD患者（尤其是未接受血液透析治疗者）外周血辅助性T淋巴细胞数量减少且相关细胞因子分泌异常；同时，患者处于氧化应激状态，可能是通过诱导CD4+T细胞过度凋亡和Th1/Th2细胞因子失衡使机体免疫功能缺陷进一步加重。

关键词: 终末期肾病, 辅助性T淋巴细胞, 细胞因子, 氧化应激

Abstract:

Objective To investigate changes of helper T lymphocytes in peripheral blood and parameters of oxidative stress of patients with end-stage renal disease (ESRD) and to explore correlation between immune function and oxidative stress. Methods Forty ESRD patients were divided into hemodialysis group (HD group, n=20) and nonhemodialysis group （NHD group, n=20) based on receiving or not receiving hemodialysis. Twenty healthy volunteers over the same period served as normal control group (NC group). Vein blood samples of all patients and subjects were collected. T lymphocytes were counted and cell subsets were selected by using flow cytometry. Percentages of T lymphocyte (CD3+ T cell) and helper T lymphocyte (CD4+ T cell) in peripheral blood and ratio of CD4+ T to another T cell subset (CD8+ T cell) (CD4+/CD8+) were calculated. Rates of proliferation and apoptosis of CD4+ T cell were measured. ELISA was used to detect contents of cytokines, interferon gamma (IFN-γ) and interleukin-4 (IL-4), in cell culture medium. Parameters of oxidative stress, superoside dismutase (SOD) and malondialdehyde (MDA), were determined by xanthinoxidase method and thiobarbituric acid spectrophotometric method,respectively. The correlations between indexes were analyzed. Results Compared with NC group, percentages of CD3+ T cell and CD4+ T cell in peripheral blood and ratio of CD4+/CD8+ in HD and NHD groups decreased significantly, accompanying increased apoptosis and decreased proliferation of CD4+T cell, increased level of IL-4, decreased level of IFN-γ, decreased level of SOD, and increased level of MDA (P＜0.05 or P＜0.01). Percentages of CD3+ and CD4+ T cells, apoptosis rate of CD4+ T cell, contents of IL-4 and IFN-γ had significant differences between HD and NHD groups (P＜0.05). The percentage of CD4+ T cell in ESRD patients was negatively correlated with the apoptosis (r=-0.87，P＜0.01), and positively correlated with the proliferation (r=0.73，P＜0.01). The rate of apoptosis of CD4+ T cell was positively correlated with the level of MDA (r=0.58, P＜0.01), and negatively correlated with SOD (r=-0.51，P＜0.01). The level of IL-4 was positively correlated with the level of MDA (r=0.47，P＜0.01). Conclusion The ESRD patients, especially those not receiving hemodialysis, had reduction in helper T lymphocytes in peripheral blood and abnormal secretion of relative cytokines. The excessive apoptosis of CD4+T cell and Th1/Th2 cytokine imbalance may be induced by oxidative stress, which aggravates immune functional defect.

Key words: end-stage renal disease, helper T lymphocyte, cytokines, oxidative stress