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目的 检测活动性狼疮肾炎 (LN)患者外周血单个核细胞 (PBMC)钙调磷酸神经酶(calcineurin ,CaN)活性及其与PBMC分泌免疫球蛋白、自身抗体的关系 ,探讨FK5 0 6治疗狼疮肾炎的作用机制。方法 体外培养活动性LN患者PBMC ,应用发色底物法检测胞质CaN活性 ,酶联免疫吸附法 (ELISA)测定细胞培养上清免疫球蛋白和抗dsDNA抗体。结果 ①单纯培养时 ,LN组CaN活性显著高于对照组 [(4 8 6± 4 7)nmol/mg蛋白vs(8 9± 2 7)nmol/mg蛋白 ,P <0 0 0 1 ];在PMA +Ionomycin刺激下 ,各组CaN活性均升高 ,LN组CaN活性明显高于正常对照组 [(71 2± 1 2 9)nmol/mg蛋白vs (34 2± 8 4 )nmol/mg蛋白 ,P <0 0 0 1 ];②单纯培养时 ,LN组PBMC培养上清中IgG浓度显著高于对照组 [(2 1 0 8± 6 0 0 )mg/Lvs (1 4 97± 5 1 6 )mg/L ,P <0 0 5 ];刺激条件下 ,LN组IgG水平显著高于对照组 [(4 991± 1 2 0 2 )mg/Lvs (3374± 1 1 6 6 )mg/L ,P <0 0 0 1 ];③单纯培养时 ,LN组PBMC培养上清中抗dsDNA抗体水平显著高于对照组 [(1 37± 0 1 6 )BIvs (0 71± 0 0 5 )BI,P <0 0 5 ];刺激条件下 ,LN组抗dsDNA抗体水平显著高于对照组 [(2 38± 1 1 7)vs (1 0 9±0 0 2 )BI ,P <0 0 0 1 ];④CaN特异的拮抗剂FK5 0 6显著抑制LNPBM
Objective To investigate the activity of calcineurin (CaN) in peripheral blood mononuclear cells (PBMC) and the relationship between the activity of PBMC and the secretion of immunoglobulin and autoantibodies in patients with active lupus nephritis (LN). To investigate the effects of FK506 on lupus nephritis The mechanism of action. Methods The PBMC of patients with active LN were cultured in vitro. The activity of cytoplasmic CaN was detected by chromogenic substrate method. The immunoglobulin and anti-dsDNA antibody of cell culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA). Results ① In pure culture, the activity of CaN in LN group was significantly higher than that in control group [(486 ± 47) nmol / mg protein vs (89 ± 27) nmol / mg protein, P <0.01] The activity of CaN in LN group was significantly higher than that in the control group [(71 2 ± 122 9 nmol / mg protein vs 34 2 ± 8 4 nmol / mg protein, PMA + Ionomycin stimulation, P <0 0 0 1]; ② In pure culture, the IgG concentration in the culture supernatant of PBMC in LN group was significantly higher than that in control group [(21 0 ± 80) mg / L vs (1497 ± 5 1 6) mg / L, P <0.05). Under stimulation, the level of IgG in LN group was significantly higher than that in control group [(4 991 ± 122) mg / L vs 3374 ± 116 6 mg / L, P <0 0 0 1]. ③ In pure culture, the level of anti-dsDNA antibody in the supernatant of PBMC of LN group was significantly higher than that of the control group [(1 37 ± 0 1 6) BIvs (0 71 ± 0 0 5) BI, P < 0 0 5]. Under stimulation, the level of anti-dsDNA antibody in LN group was significantly higher than that in control group [(2 38 ± 1 1 7) vs (109 ± 0 0 2) BI, P 0 01 0; The specific antagonist FK5 0 6 significantly inhibited LNPBM