不同分期慢性肾脏疾病患者γ干扰素和白细胞介素4基因表达水平与功能变化  

Gene expression and functional changes of interferon-γ/interleukin-4 in patients with different stage of chronic kidney disease

作　　者：张丽红[1] 袁瑾[1] 周韵娇[1] 牛建英[1] 

机构地区：[1]复旦大学附属上海市第五人民医院肾内科,上海200240

出　　处：《国际移植与血液净化杂志》2014年第2期31-34,共4页International Journal of Transplantation and Hemopurification

基　　金：复旦大学985医院优势学科项目，上海市闵行区科委资助项目，上海市闵行区卫生局重点专科课题资助项目

摘　　要：目的 通过观察慢性肾脏疾病患者外周血淋巴细胞细胞因子γ干扰素、白细胞介素4 mRNA合成及蛋白分泌水平,探讨慢性肾功能衰竭时免疫功能紊乱的发生机制.方法 入选慢性肾功能不全患者18例（慢性肾脏疾病3、4、5期各6例）和健康对照者4名,于肘静脉采血10ml,采用含有15％小牛血清（56℃水浴中灭活30 min）的RPMI 1640液培养淋巴细胞,应用TRIzon法提取总RNA,以RT-PCR法对γ干扰素和白细胞介素4mRNA进行扩增,琼脂糖凝胶进行电泳分离,双抗体夹心法检测γ干扰素和白细胞介素4蛋白浓度.结果 慢性肾脏疾病4、5期者外周血淋巴细胞培养上清液中γ干扰素浓度低于健康对照者和慢性肾脏疾病3期者,健康对照组、慢性肾脏疾病3、4、5期患者白细胞介素4浓度差异无统计学意义,慢性肾脏疾病4期者γ干扰素/白细胞介素4比值最低;慢性肾脏疾病4、5期者γ干扰素mRNA的相对积分光密度低于健康对照组和慢性肾脏疾病3期者,慢性肾脏疾病5期者白细胞介素4 mRNA的积分光密度值高于健康对照组和慢性肾脏疾病3、4期者,慢性肾脏疾病4、5期者γ干扰素与白细胞介素4mRNA积分光密度的比值低于健康对照组和慢性肾脏疾病3期者.结论 慢性肾脏疾病患者确实存在T淋巴细胞功能异常.随着慢性肾脏疾病的进展,T淋巴细胞分泌细胞因子的功能呈下降趋势.在慢性肾脏疾病的发生和发展中,Th1/Th2平衡存在向TH2偏移的趋势.Objective To investigate gene expression and functional change of interferon-γ（IFN-γ）/interleukin-4 （IL-4） in patients with different stage of chronic kidney disease （CKD）.Methods Eighteen patients with chronic renal failure （CRF） and 4 healthy controls were enrolled in this study.RPMI 1640containing 15% fetal calf serum （56 ℃ water bath for 30 minutes） was used for lymphocyte culture.Total RNA was extracted by using TRIzon method,and RT-PCR was conducted for IFN-γ and IL-4 mRNA amplification.Double antibody sandwich was used to detect IFN-γand IL-4 protein.Results Concentration of IFN-γand IL-4 in the supematants of peripheral blood lymphocytes was higher in CKD 4 and 5 patients than in CKD 3 or healthy controls.Moreover,lower relative IOD value of IFN-γ mRNA was found in CKD 4 or 5 patients.In comparison with healthy controls and CKD 3 or 4 subjects,those of CKD 5 bad higher relative IOD value of IL-4 mRNA.IODIL-4 mRNA/IODIFN-γmRNA was increased in CKD 4 and 5 individuals.Conclusion Our investigation confirm abnormal T cell function in CKD patients,and T cell cytokine secretion function show a downward trend as CKD progress.

关 键 词：慢性肾脏疾病 Γ干扰素 白细胞介素4 INTERFERON-Γ INTERLEUKIN-4 

分 类 号：R692.5[医药卫生—泌尿科学]