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目的 了解磷脂酰肌醇三激酶 (PI3 K)在狼疮性肾炎 (LN)外周血单个核细胞 (PBMC)中的活化情况。方法 2 5例狼疮性肾炎患者被分为活动期 (1 3例 )和非活动期 (1 2例 ) ,并以 1 2例健康人为对照。PBMC提取采用Ficol密度梯度离心法 ,PBMCPI3 K酪氨酸磷酸化产物表达采用免疫沉淀、免疫印迹法。细胞培养上清IgG和抗dsDNA检测采用微量ELISA法。结果 与对照组相比 ,活动性SLE患者PI3 K酪氨酸磷酸化活性增高 (P <0 0 1 ) ,非活动性SLE患者PI3 K酪氨酸磷酸化活性无显著性差异 (P>0 0 5 ) ;相关分析显示 ,PI3 K磷酸化产物表达与狼疮活动指数 (r =0 6 1 7,P <0 0 1 )、血清抗dsFNA(r=0 5 4 4,P <0 0 1 )、免疫球蛋白IgG(r=0 6 91 ,P <0 0 1 )呈明显正相关关系 ,与血清补体C3呈明显负相关关系 (r=- 0 6 1 7,P <0 0 1 )。PI3 K特异抑制剂Wortamannin能抑制CD3mAb诱导的SLEPBMC抗dsDNA和IgG的产生 (P <0 0 1 )。结论 PI3 K过度活化可能参与了SLE自身抗体的产生 ,检测PI3 K活化状况可能有助于临床狼疮活动的判断。
Objective To understand the activation of phosphatidylinositol tri-kinase (PI3K) in peripheral blood mononuclear cells (PBMCs) of lupus nephritis (LN). Methods Twenty-five patients with lupus nephritis were divided into active stage (13 cases) and inactive stage (12 cases), and 12 healthy controls were used as control. The PBMCs were extracted by Ficol density gradient centrifugation. PBMCPI3K tyrosine phosphorylation products were expressed by immunoprecipitation and immunoblotting. Cell culture supernatant IgG and anti-dsDNA detection using micro ELISA. Results Compared with the control group, the PI3K tyrosine phosphorylation activity was increased in patients with active SLE (P <0.01), while there was no significant difference in PI3K tyrosine phosphorylation activity in inactive SLE patients (P> 0 0 5). Correlation analysis showed that the phosphorylation of PI3K was correlated with lupus activity index (r = 0 6 1 7, P 0 01), serum anti-dsFNA (r 0 0544, P 0 01) Immunoglobulin IgG (r = 0 6 91, P 0 01) showed a significant positive correlation, and negatively correlated with serum complement C3 (r = - 0 6 1 7, P 0 01). PI3K-specific inhibitor Wortamannin inhibited CD3 mAb-induced SLEPBMC anti-dsDNA and IgG production (P <0.01). Conclusion PI3K overexpression may be involved in the production of SLE autoantibodies. Detecting the activation of PI3K may contribute to the judgment of clinical lupus activity.