【摘要】 目的 探讨褪黑素对狼疮性肾炎（LN）病人血清诱导的肾小球内皮细胞（HRGEC）炎症的影响。方法 将不同剂量的褪黑素（10、100、250、500、750、1000μmol/L）加入HRGEC培养体系中24 h，应用CCK-8测定细胞活力。采集抗双链DNA阳性（抗dsDNA阳性）的10例LN病人血清刺激HRGEC，应用Western blot检测HRGEC中NOD样受体热蛋白结构域蛋白3(NLRP3)炎症小体活化、ELISA测定细胞上清液中IL-1β水平、DCFH-DA和DHE检测细胞活性氧（ROS）的产生，利用免疫荧光法检测NF-κB/p65的核转位。结果 CCK-8测定结果提示≥750μmol/L的褪黑素显著降低HRGEC细胞活力（P<0.05）。褪黑素抑制人LN血清诱导的HRGEC中NLRP3炎症小体激活及下游的IL-1β产生（P<0.05）；褪黑素逆转人LN血清刺激的HRGEC细胞ROS产生（P<0.05）；褪黑素抑制人LN血清诱导的HRGEC细胞NF-κB活性。结论 褪黑素可能通过抑制炎症反应改善LN病人血清导致的肾小球内皮细胞损伤，它可作为LN的一种潜在的治疗药物。更多还原

【Abstract】 Objective To explore the effect of melatonin on lupus nephritis(LN)serum-induced inflammation in human renal glomerular endothelial cell(HRGEC). Methods Different doses of melatonin(10,100,250,500,750,1000 μmol/L)were added to HRGEC culture system for 24 hours,and cell viability was measured by cell Counting Kit-8(CCK-8). Anti-dsDNA-positive serum was collected from LN patients(n=10)to stimulate HRGEC.NLRP3 inflammasome activation in HRGEC was detected by Western blot;IL-1β level in cell supernatant was determined by ELISA;reactive oxygen species(ROS)production was detected by DCFH-DA and DHE;and nuclear translocation of NF-κB/p65 was detected by immunofluorescence. Results The CCK-8 assay indicated that melatonin ≥750 μmol/L significantly decreased the viability of HRGEC(P<0.05). Melatonin inhibited the activation of NLRP3 inflammasome and downstream IL-1β production in HRGEC induced by human LN serum(P<0.05). Melatonin reversed human LN serum induced ROS production in HRGEC(P<0.05). Melatonin inhibited NF-κB activity induced by human LN serum in HRGEC. Conclusion Melatonin may ameliorate glomerular endothelial cell injury caused by serum in LN by inhibiting inflammatory response,and it may be a potential therapeutic agent for LN.更多还原