中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2015年
7期
521-526
,共6页
王秋红%封晓娟%吴超%刘淑霞
王鞦紅%封曉娟%吳超%劉淑霞
왕추홍%봉효연%오초%류숙하
狼疮肾炎%高迁移率族蛋白1%细胞增殖%增殖细胞核抗原%肾功能
狼瘡腎炎%高遷移率族蛋白1%細胞增殖%增殖細胞覈抗原%腎功能
랑창신염%고천이솔족단백1%세포증식%증식세포핵항원%신공능
Lupus nephritis%High mobility group box chromosomal protein 1%Cell proliferation%Proliferating cell nuclear antigen%Renal function
目的 通过敲低肾组织高迁移率族蛋白1(HMGB1)表达,探讨其对改善狼疮肾炎小鼠肾功能,降低肾小球细胞增殖水平的影响.方法 MRL/Faslpr鼠(n=24)被随机分为模型组、shHMGB1组和空质粒组;选取周龄、体质量相匹配的MRL/MpJ鼠为健康对照组.shHMGB1组和空质粒组采用电穿孔转染技术分别转染shHMGB1质粒和空质粒,模型组和健康对照组仅转染生理盐水.用全自动生化分析仪检测小鼠血清尿素氮和肌酐水平,测定尿蛋白浓度并计算24 h尿蛋白量(UP).HE染色观察肾组织的形态学表现;免疫荧光和Western印迹法检测小鼠肾小球中HMGB1和增殖细胞核抗原(PCNA)的表达;实时定量PCR法检测小鼠肾小球HMGB1和PCNA mRNA表达变化.结果 (1)与健康对照组相比,模型组小鼠肾小球HMGB1 mRNA和蛋白的表达升高(均P＜0.05);与模型组相比,shHMGB1组小鼠肾小球中HMGB1 mRNA和蛋白的表达降低(均P＜0.05).(2)与模型组相比,shHMGB1组小鼠尿蛋白减少(P＜0.05).(3)免疫荧光和Western印迹结果显示,与健康对照组相比,模型组小鼠肾小球中PCNA mRNA和蛋白表达升高(P＜0.05).与模型组相比,shHMGB1组肾小球中PCNA表达降低(P＜0.05).结论 敲低肾组织HMGB1表达可改善狼疮肾炎小鼠的肾功能,降低肾小球细胞的增殖水平.
目的 通過敲低腎組織高遷移率族蛋白1(HMGB1)錶達,探討其對改善狼瘡腎炎小鼠腎功能,降低腎小毬細胞增殖水平的影響.方法 MRL/Faslpr鼠(n=24)被隨機分為模型組、shHMGB1組和空質粒組;選取週齡、體質量相匹配的MRL/MpJ鼠為健康對照組.shHMGB1組和空質粒組採用電穿孔轉染技術分彆轉染shHMGB1質粒和空質粒,模型組和健康對照組僅轉染生理鹽水.用全自動生化分析儀檢測小鼠血清尿素氮和肌酐水平,測定尿蛋白濃度併計算24 h尿蛋白量(UP).HE染色觀察腎組織的形態學錶現;免疫熒光和Western印跡法檢測小鼠腎小毬中HMGB1和增殖細胞覈抗原(PCNA)的錶達;實時定量PCR法檢測小鼠腎小毬HMGB1和PCNA mRNA錶達變化.結果 (1)與健康對照組相比,模型組小鼠腎小毬HMGB1 mRNA和蛋白的錶達升高(均P＜0.05);與模型組相比,shHMGB1組小鼠腎小毬中HMGB1 mRNA和蛋白的錶達降低(均P＜0.05).(2)與模型組相比,shHMGB1組小鼠尿蛋白減少(P＜0.05).(3)免疫熒光和Western印跡結果顯示,與健康對照組相比,模型組小鼠腎小毬中PCNA mRNA和蛋白錶達升高(P＜0.05).與模型組相比,shHMGB1組腎小毬中PCNA錶達降低(P＜0.05).結論 敲低腎組織HMGB1錶達可改善狼瘡腎炎小鼠的腎功能,降低腎小毬細胞的增殖水平.
목적 통과고저신조직고천이솔족단백1(HMGB1)표체,탐토기대개선랑창신염소서신공능,강저신소구세포증식수평적영향.방법 MRL/Faslpr서(n=24)피수궤분위모형조、shHMGB1조화공질립조;선취주령、체질량상필배적MRL/MpJ서위건강대조조.shHMGB1조화공질립조채용전천공전염기술분별전염shHMGB1질립화공질립,모형조화건강대조조부전염생리염수.용전자동생화분석의검측소서혈청뇨소담화기항수평,측정뇨단백농도병계산24 h뇨단백량(UP).HE염색관찰신조직적형태학표현;면역형광화Western인적법검측소서신소구중HMGB1화증식세포핵항원(PCNA)적표체;실시정량PCR법검측소서신소구HMGB1화PCNA mRNA표체변화.결과 (1)여건강대조조상비,모형조소서신소구HMGB1 mRNA화단백적표체승고(균P＜0.05);여모형조상비,shHMGB1조소서신소구중HMGB1 mRNA화단백적표체강저(균P＜0.05).(2)여모형조상비,shHMGB1조소서뇨단백감소(P＜0.05).(3)면역형광화Western인적결과현시,여건강대조조상비,모형조소서신소구중PCNA mRNA화단백표체승고(P＜0.05).여모형조상비,shHMGB1조신소구중PCNA표체강저(P＜0.05).결론 고저신조직HMGB1표체가개선랑창신염소서적신공능,강저신소구세포적증식수평.
Objective To investigate the effect of high mobility group box chromosomal protein 1 (HMGB1) knockdown on improving renal function and decreasing cell proliferation of glomeruli in lupus nephritis (LN) MRL/Faslpr mice.Methods Twenty-four MRL/Faslpr mice were randomly divided into 3 groups:LN model group,shHMGB1 group and empty plasmid group.Besides,eight MRL/MpJ mice,age and mass matched to the MRL/Faslpr mice,were chosen as normal control group (shNC group).Electroporation technology was used for in vivo transfection in treatment group.shHMGB1 group and empty plasmid group were transfected by electroporation technology for shHMGB1 plasmids and empty plasmid,LN model group and normal control group were transfected only with saline.Automatic biochemical analyzer was used to detect serum urea nitrogen (BUN) and creatinine (Scr) levels and 24 h urinary protein (UP) was tested.HE staining was used to detect the pathological change of renal tissues; real-time PCR,immunofluorence staining and Western blotting were used to detect the mRNA and protein expression of HMGB1 and PCNA.Results (1) The HMGB1 mRNA and protein expression in LN group increased compared with those in control group,HMGB1 mRNA and protein expression in shHMGB1 group reduced compared with those in LN model group (all P ＜ 0.05).(2) 24 h UP of MRL/Faslpr mice in shHMGB1 group significantly reduced compared with those in LN group (P ＜ 0.05).(3) Immunofluorence and Western blotting showed that positive signal of proliferating cell nuclear antigen (PCNA) was mainly located in nuclei,PCNA mRNA and protein in glomeruli of LN model group increased compared with those of control mice (P ＜ 0.05).Interestingly,PCNA expression in glomeruli of shHMGB1 group remarkably reduced (P ＜ 0.05).Conclusions shHMGB1 significantly improves renal function and decreases cell proliferation of glomeruli in LN MRL/Faslpr mice.
