【摘要】 目的观察小鼠肾小管上皮细胞(TEC)在白介素4(IL-4)及CD40激活状态下钙调神经磷酸酶(CaN)活性,并进一步探讨其与趋化因子RANTES分泌的关系。方法取Ⅱ级6～7周的雌性DBA小鼠,分离肾小管上皮细胞进行培养、刺激。采用RT-PCR检测RANTESmRNA表达;ELISA检测培养上清中RANTES含量;流式细胞仪检测CD40表达。结果(1)常规培养TEC可表达一定量的CD40;用IL-4刺激TEC24h,细胞表面CD40平均荧光强度(MFI)显著高于对照组(7.92±0.56vs4.33±0.30,P<0.001)。(2)常规培养TEC中有一定量的CaN活化(8.98±0.56)nmol/mg.pro;用IL-4、CD40抗体(mAb)及IL-4+CD40mAb刺激细胞,3组CaN活性均显著高于对照组(P<0.05)。(3)常规培养TEC仅分泌极少量的RANTES;在IL-4刺激或CD40mAb激活后TECRANTES蛋白分泌增高,分别为(43.61±13.73)pg/ml和(73.77±4.28)pg/ml,显著高于对照组(14.78±2.20)pg/ml(P<0.001)。(4)常规培养TEC微量表达RANTESmRNA;用IL-4、CD40mAb及IL-4+CD40mAb刺激细胞24h,各刺激组RANTESmRNA表达显著高于对照组(P<0.05)。(5)在IL-4、CD40mAb及IL-4+CD40mAb刺激下,FK506对TEC分泌RANTES蛋白及mRNA有显著抑制作用(P均<0.05)。结论Th2细胞因子IL-4及CD40-CD40配体(CD40L)共刺激信号可通过活化TECCaN,调控RANTES基因表达及分泌、更多还原

【Abstract】 Objective To investigate whether calcineurin(CaN)is activated in murine renal tubular epithelial cells (TEC)and the effect of IL-4, CD40L on RANTES production and the CaN activity. Methods TECs were obtained from DBA mous e and cultured in conditioned medium. Expression of RANTES mRNA in TEC was measu red by RT-PCR. Content of RANTES in supernatant of cultured TEC was detected by ELISA. Expression of CD40 in TEC was examined by flow cytometry. Results (1)The expression of CD40 in TEC was clearly demonstrated. Activation of TEC by IL-4 resulted in significant increase of CD40 expression compared to control TEC(7.92 ±0.56 vs 4.33±0.30,P< 0.001).(2)Nonstimulated control TEC showed low CaN acti vity, which was significantly enhanced by the stimulation of IL-4,CD40 or combi ned stimulation of IL-4 and CD40mAb repectively(P< 0.05). (3)Low level of RANTE S was detected by ELISA in supernatants of murine TEC without stimulation. TEC s timulated with either IL-4 or CD40mAb resulted in strong induction of RANTES pr oduction up to (43.61±13.73) or (73.77±4.28)pg/ml respectively, which was stat istically higher than (14.78±2.20)pg/ml in control. The difference of RANTES be tween above two stimulation groups and control group was significant(P< 0.01). ( 4)RANTES mRNA was expressed at basal level in control TEC, and this expressing l evel increased significantly when TECs were stimulated with IL-4、CD40 or IL-4 in combination with CD40mAb respectively. (5)Addition of FK506 could block the stimulation of IL-4 and CD40mAb,and reduce the production of RANTES, and the expression of RANTES mRNA significantly. Conclusions The incre ase of CaN activity by the stimulation of IL-4 and CD40mAb on TEC upregulates t he RANTES production. CaN activation may participate in the inflammation of TEC.更多还原