狼疮肾炎患者外周血单个核细胞钙调神经磷酸酶活性的检测及CD40L表达变化  

SIGNIFICANCE OF CALCINEURIN ACTIVATION AND CD40L EXPRESSION IN PATIENTS WITH ACTIVE LUPUS NEPHRITIS

作　　者：顾海峪[1] 梁鸣[1] 卢建华[1] 阳晓[2] 李幼姬[2] 孔庆喻[2] 许韩师[2] 余学清[2] 

机构地区：[1]广州市第一人民医院急诊科,广东广州510180 [2]中山大学附属第一医院,广东广州510080

出　　处：《中国应用生理学杂志》2005年第3期334-338,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金(30271668);广东省医学科研基金(B2003122);广东省中医药局科研基金资助项目(403029)

摘　　要：目的:检测活动性狼疮肾炎(LN)患者外周血单个核细胞(PBMC)钙调神经磷酸酶(calcineurin,CaN)活性及其与PBMCCD40L表达的关系。方法:体外培养活动性LN患者PBMC,应用发色底物法检测胞浆CaN活性,流式细胞仪检测细胞CD40L的表达。结果:①在单纯培养情况下,正常对照组和LN组PBMC均出现一定量CaN活化,活动性LN组显著高于正常对照组(46.08±5.58nmol/mgprovs8.81±3.61nmol/mgpro,P<0.01);在PMA+Ionomycin刺激下,各组CaN活性均升高,活动性LN组CaN活性明显高于正常对照组(69.34±12.59nmol/mgprovs37.12±11.57nmol/mgpro,P<0.01);②LN患者PBMC在单纯培养和PMA+Ionomycin刺激时CD40L蛋白和mRNA表达均显著高于相应的对照组(P<0.01);③在单纯培养和PMA+Ionomycin刺激时,FK506对LNPBMC表达CD40L蛋白和mRNA均有显著抑制作用(P<0.01)。结论:LN患者PBMC存在CaN过度活化;LN患者PBMC高效表达CD40L与其CaN过度活化密切相关,通过阻断CaN活化可调控CD40-CD40L共刺激信号途径的活化。Aim： To investigate the significance of the calcineurin（CaN） activation in active lupus nephritis patient. Methods： Peripheral blood mononuclear cells（PBMCs） were separated from twenty-one active LN patients and 12 healthy controls. Phosphatase activity of CaN was determined using the CaN ashy kit by measuring the content of released PO4. Reverse transcription-PCR was used to detect the expression of CD40L mRNA. Flow cytometry analysis was used to detect the expression of CD40L in LN PBMC. Results： ① Increased activation of CaN in spontaneous cultured PBMC in active LN group was found as compared with control group（46.08 ± 5.58 vs 8.81 ± 3.61, P〈 0.01 ）. In stimulated by PMA/Ionomycin , activity of CaN in active LN group was also higher than that of control（69.34 ± 12.59 vs 37.12 ± 11 . 57, P 〈 0.01 ）. ② Relative content of CD40L in PBMC in active LN groups increased significantly as compared with the control groups under spontaneous and PMA/Ionomycin-induced culture, respectively （ P 〈 0.01 ）. ③ FK506 reduced significantly production of CD40L in spontaneous and PMA/Ionomycin-induced PBMC of LN. Conclusion： Elevated activation of CaN in active LN may participate in regulation overexpression of CD40L in PBMC of LN. Through inhibiting CaN activity , FK506 may prevent abnormal activation of CD40-CD40L costimulatory pathway in lupus nephritis.

关 键 词：狼疮肾炎 外周血单个核细胞 钙调神经磷酸酶 CD40配体 

分 类 号：R593.24[医药卫生—内科学]