中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2013年
4期
444-447
,共4页
黄春%杨明%施晓芸%陈晓春
黃春%楊明%施曉蕓%陳曉春
황춘%양명%시효예%진효춘
缺氧%糖基化终产物,高级%早期生长反应蛋白质
缺氧%糖基化終產物,高級%早期生長反應蛋白質
결양%당기화종산물,고급%조기생장반응단백질
Anoxia%Glycosylation end-products,advanced%Early growth response protein
目的 探讨急性缺氧对早期生长反应因子-1(Egr-1)和单核细胞趋化蛋白-1(MCP-1)表达的影响及其可能的信号机制. 方法 取整体缺氧小鼠模型的主动脉,实时定量反转录聚合酶链反应(RT-PCR)检测Egr-1和MCP-1 mRNA含量、蛋白印迹检测Egr-1和晚期糖基化终末产物受体(RAGE)抗原表达,凝胶迁移或电泳迁移率实验(EMSA)检测Egr-1 DNA结合活性；以可溶性RAGE(sRAGE)阻断RAGE信号后,观察其对缺氧诱导Egr-1表达的影响. 结果 缺氧30 min,Egr-1mRNA表达上调,为对照组的(28.3±0.9)倍(F=617.17,P＜0.01)；缺氧45 min,Egr-1的抗原含量为对照组的(5.7±0.3)倍(F=57.18,P＜0.01),Egr-1 DNA结合活力高于对照组,并被抗-Egr-1 IgG所抑制；缺氧4h,MCP-1 mRNA含量为对照组的(4.0±0.3)倍(F=30.68,P＜0.01)；缺氧15 min,RAGE抗原含量增加,sRAGE预处理减少缺氧诱导的Egr-1表达(3.3±0.2)倍与(1.4±0.2)倍(F=30.20,P＜0.01). 结论 小鼠整体缺氧诱导主动脉Egr-1及MCP-1表达上调,阻断RAGE信号显著抑制缺氧诱导的Egr-1高表达.
目的 探討急性缺氧對早期生長反應因子-1(Egr-1)和單覈細胞趨化蛋白-1(MCP-1)錶達的影響及其可能的信號機製. 方法 取整體缺氧小鼠模型的主動脈,實時定量反轉錄聚閤酶鏈反應(RT-PCR)檢測Egr-1和MCP-1 mRNA含量、蛋白印跡檢測Egr-1和晚期糖基化終末產物受體(RAGE)抗原錶達,凝膠遷移或電泳遷移率實驗(EMSA)檢測Egr-1 DNA結閤活性；以可溶性RAGE(sRAGE)阻斷RAGE信號後,觀察其對缺氧誘導Egr-1錶達的影響. 結果 缺氧30 min,Egr-1mRNA錶達上調,為對照組的(28.3±0.9)倍(F=617.17,P＜0.01)；缺氧45 min,Egr-1的抗原含量為對照組的(5.7±0.3)倍(F=57.18,P＜0.01),Egr-1 DNA結閤活力高于對照組,併被抗-Egr-1 IgG所抑製；缺氧4h,MCP-1 mRNA含量為對照組的(4.0±0.3)倍(F=30.68,P＜0.01)；缺氧15 min,RAGE抗原含量增加,sRAGE預處理減少缺氧誘導的Egr-1錶達(3.3±0.2)倍與(1.4±0.2)倍(F=30.20,P＜0.01). 結論 小鼠整體缺氧誘導主動脈Egr-1及MCP-1錶達上調,阻斷RAGE信號顯著抑製缺氧誘導的Egr-1高錶達.
목적 탐토급성결양대조기생장반응인자-1(Egr-1)화단핵세포추화단백-1(MCP-1)표체적영향급기가능적신호궤제. 방법 취정체결양소서모형적주동맥,실시정량반전록취합매련반응(RT-PCR)검측Egr-1화MCP-1 mRNA함량、단백인적검측Egr-1화만기당기화종말산물수체(RAGE)항원표체,응효천이혹전영천이솔실험(EMSA)검측Egr-1 DNA결합활성；이가용성RAGE(sRAGE)조단RAGE신호후,관찰기대결양유도Egr-1표체적영향. 결과 결양30 min,Egr-1mRNA표체상조,위대조조적(28.3±0.9)배(F=617.17,P＜0.01)；결양45 min,Egr-1적항원함량위대조조적(5.7±0.3)배(F=57.18,P＜0.01),Egr-1 DNA결합활력고우대조조,병피항-Egr-1 IgG소억제；결양4h,MCP-1 mRNA함량위대조조적(4.0±0.3)배(F=30.68,P＜0.01)；결양15 min,RAGE항원함량증가,sRAGE예처리감소결양유도적Egr-1표체(3.3±0.2)배여(1.4±0.2)배(F=30.20,P＜0.01). 결론 소서정체결양유도주동맥Egr-1급MCP-1표체상조,조단RAGE신호현저억제결양유도적Egr-1고표체.
Objective To investigate the impact of hypoxia on the expression of early growth response-1 (Egr-1) and monocyte chemoattractant protein-1 (MCP-1) in mouse aorta,and to probe the underlying mechanism involving receptor for advanced glycation end-products (RAGE).Methods 3-month-old C57BL/6 mice were subjected to hypoxia [(6.0±0.5) ％ oxygen] to establish the global hypoxia model(n=6 rats for each).Aortas were dissected,Egr-1 mRNA and MCP-1 mRNA were detected by real time RT-PCR,Egr-1 and RAGE proteins were tested by Western blot,and Egr-1 DNA binding activity was assayed by electrophoretic mobility shift assay (EMSA).For blockade of RAGE,mice were pretreated with soluble RAGE (sRAGE) for 1 h by intra-peritoneal injection before they were exposed to hypoxia.Mice with normoxia were used as controls.Results After 30 minutes of hypoxic exposure,Egr-1 mRNA in aorta was increased to (28.3±0.9)folds compared with normoxic controls (F=617.17,P＜0.01),and the induction persisted for at least 3 hours.After 45 minutes of hypoxic exposure,Egr-1 proteins in aorta was increased to (5.7 ± 0.3) folds compared with normoxic controls (F =57.18,P＜ 0.01); the enhanced DNA binding activity of Egr-1 by hypoxia was attenuated by pretreatment with anti-Egr-1 lgG.After 4 hours of hypoxic exposure,MCP-1 mRNA expression in aorta was increased to(4.0±0.3)folds compared with normoxic controls (F=30.68,P＜0.01).RAGE antigen was increased significantly within 30 minutes of hypoxic exposure,with the peak at 15 minutes; hypoxia-induced Egr-1 mRNA expression was significantly attenuated by pretreatment with sRAGE (3.3 ± 0.2) folds compared with normoxic controls (F =30.20,P＜0.01).Conclusions Hypoxia significantly induces Egr-1 and MCP-1 upregulation expressions in mouse aorta,and blockade of RAGE significantly attenuates hypoxia-induced Egr-1 expression.Thcsc findings suggest RAGE signaling is involved in hypoxia-induced vascular inflammatory stress,and highlight this receptor as a potential therapeutic target to protect tissues injured by hypoxia.
