CAJ | 학술논문

目的研究高迁移率族蛋白1(high mobility group protein box 1,HMGB1)在系统性红斑狼疮肾损害中的作用.方法 ELISA检测12例健康对照、16例系统性红斑狼疮(systemic lupus erythematosus,SLE)无肾脏损害和14例合并肾脏损害的系统性红斑狼疮患者(lupus nephritis,LN)血清中HMGB1的表达情况.将体外培养的人系膜细胞分为正常对照组和HMGB1刺激组,于培养6、12、24 h后收集细胞,免疫细胞化学检测增殖细胞核抗原(PCNA)的表达变化;免疫细胞化学和流式细胞术检测Toll样受体2(TLR2)、NF-κB 065蛋白的表达变化.结果血清中HMGB1蛋白在LN组明显高于SLE组和健康对照组,且血清中HMGB1水平与LN患者的蛋白尿呈显著正相关;人重组HMGB1能够促进系膜细胞增殖;与正常对照组相比,HMGB1刺激组中TLR2、NF-κB p65蛋白表达增强;TLR2蛋白与NF-κB p65蛋白表达呈显著正相关(r=0.658,P=0.000);NF-κB p65蛋白表达与PCNA阳性表达率呈显著正相关(r=0.536,P=0.007).结论 HMGB1是狼疮性肾炎发病中的重要的细胞因子之一,可能部分通过与其受体蛋白TLR2结合激活NF-κB促进系膜细胞增生,从而引起肾脏损害.
목적 연구고천이솔족단백1(high mobility group protein box 1,HMGB1)재계통성홍반랑창신손해중적작용.방법 ELISA검측12례건강대조、16례계통성홍반랑창(systemic lupus erythematosus,SLE)무신장손해화14례합병신장손해적계통성홍반랑창환자(lupus nephritis,LN)혈청중HMGB1적표체정황.장체외배양적인계막세포분위정상대조조화HMGB1자격조,우배양6、12、24 h후수집세포,면역세포화학검측증식세포핵항원(PCNA)적표체변화;면역세포화학화류식세포술검측Toll양수체2(TLR2)、NF-κB 065단백적표체변화.결과 혈청중HMGB1단백재LN조명현고우SLE조화건강대조조,차혈청중HMGB1수평여LN환자적단백뇨정현저정상관;인중조HMGB1능구촉진계막세포증식;여정상대조조상비,HMGB1자격조중TLR2、NF-κB p65단백표체증강;TLR2단백여NF-κB p65단백표체정현저정상관(r=0.658,P=0.000);NF-κB p65단백표체여PCNA양성표체솔정현저정상관(r=0.536,P=0.007).결론 HMGB1시랑창성신염발병중적중요적세포인자지일,가능부분통과여기수체단백TLR2결합격활NF-κB촉진계막세포증생,종이인기신장손해.
Objective To explore the effect of high mobility group protein box 1(HMGB1)on the renal injure of systemic lupus erythematosus(SLE).Methods The level of HMGB1 in serum from 16 patients with SLE,14 patient with lupus nephritis(LN)and 12 healthy people was measured by ELISA.Human mesangial cells were inoeulated in the dose of 104/ml.After 24 h,cells were cultured with standard medium as control group or with medium supplemented with 10μg/L human recombinant protein HMGB1 as trial group in vitro.Then cells were collected in 6,12,24 h respectively,as well as normal control group cells.Immunocytochemieal staining was adopted to detect proliferating cell nuclear antigen(PCNA)expressions on mesangial cell in diferent groups.Immunocytochemical staining and FCM were performed to analysis the changes of TLR2,NF-κB p65 proteins expressions.Results ELISA results showed that the level of HMGB1 protein in serum were higher in patients with LN than those in SLE and healthy people.HMGB1 in serum was positively collated with proteinuria in LN.Immunocytochemical staining showed the number of positive expression cells of PCNA increased in time-dependent manner.Immunocytochemical staining and FCM indicated that HMGB1 could significantly up.regulate the expression of TLR2,NF-κB p65 protein.NF-κB D65 protein Was positively correlated with the expression of TLR2 and the positive ratio of PCNA.Conclusion HMGB1 is one of the important eytokine in the pathogenesis of lupus nephritise:HMGB1 could bind to cell-surface receptors-TLR2 of mesangial cell and effectively activate NF-κB.further promote cell proliferation,which may be one of the mechanisms in renal iniare in LN.