褪黑素抑制狼疮性肾炎血清诱导的人肾小球内皮细胞炎症  

Melatonin inhibits inflammation induced by human lupus nephritis serum in human renal glomerular endothelial cells

作　　者：毛任翔[1] 李帅[2] 曹莹 赵继军 MAO Ren-xiang;LI Shuai;CAO Ying;ZHAO Ji-jun(Department of Dermatology,First Afiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Department of Biochemistry and Molecular Biology,Guangzhou Medical University,Guangzhou 511436,China;Department of Rheumatology and Immunology,First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院皮肤科,广东广州510080 [2]广州医科大学生物化学与分子生物学教研室,广东广州511436 [3]中山大学附属第一医院风湿免疫科,广东广州510080

出　　处：《解剖学研究》2023年第5期460-464,474,共6页Anatomy Research

基　　金：国家自然科学基金项目(82071820)。

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。University;Department of Rheumatology and Immunology,First Affiliated Hospital of Sun Yat-sen University;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(10,100,250,500,750,1000μmol/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-1βlevel 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≥750μmol/L significantly decreased the viability of HRGEC(P<0.05).Melatonin inhibited the activation of NLRP3 inflammasome and downstream IL-1βproduction 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.

关 键 词：狼疮性肾炎 褪黑素 肾小球内皮细胞 NOD样受体热蛋白结构域蛋白3 活性氧 核因子κB 

分 类 号：R593.242[医药卫生—内科学]