中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
32期
2274-2277
,共4页
黏着斑激酶%肌细胞,平滑肌%细胞增殖%肺动脉%细胞低氧
黏著斑激酶%肌細胞,平滑肌%細胞增殖%肺動脈%細胞低氧
점착반격매%기세포,평활기%세포증식%폐동맥%세포저양
Focal adhesion kinase%Myocytes,smooth muscle%Cell proliferation%Pulmonary artery%Cell hypoxia
目的 探讨低氧状态下黏着斑激酶(FAK)对人肺动脉平滑肌细胞(HPASMC)增殖的影响机制.方法 采用FAK寡核苷酸转染法对HPASMC进行低氧处理,免疫沉淀法测定胞质蛋白FAK、生长因子受体结合蛋白2(Grb2)及吻蛋白的活性,Western印迹法测定胞质蛋白FAK、Grb2及吻蛋白的蛋白表达,免疫组织化学法测定胞质FAK、Grb2及吻蛋白表达.结果 低氧处理后胞质FAK、Grb2及吻蛋白的活性增加.胞质FAK、Grb2及吻蛋白的蛋白表达值在低氧处理1.5 h时分别为43.4±1.4、69.7±1.9、59.3±1.6,在低氧处理24 h时分别为41.3±1.3、71.3±1.5、59.4±1.8,较常氧处理1.5 h(35.7±1.2、48.7±1.3、33.2±1.8)和24 h时(41.3±1.3、50.2±1.7、38.9±1.9)明显增加,差异均有统计学意义(均P＜0.05).免疫组织化学结果也显示低氧情况下胞质FAK、Grb2及吻蛋白的表达增强.结论 低氧状态下HPASMC增殖与胞质FAK、Grb2及吻蛋白有关.这些胞质蛋白在低氧情况下对细胞生长和分化具有调控作用.
目的 探討低氧狀態下黏著斑激酶(FAK)對人肺動脈平滑肌細胞(HPASMC)增殖的影響機製.方法 採用FAK寡覈苷痠轉染法對HPASMC進行低氧處理,免疫沉澱法測定胞質蛋白FAK、生長因子受體結閤蛋白2(Grb2)及吻蛋白的活性,Western印跡法測定胞質蛋白FAK、Grb2及吻蛋白的蛋白錶達,免疫組織化學法測定胞質FAK、Grb2及吻蛋白錶達.結果 低氧處理後胞質FAK、Grb2及吻蛋白的活性增加.胞質FAK、Grb2及吻蛋白的蛋白錶達值在低氧處理1.5 h時分彆為43.4±1.4、69.7±1.9、59.3±1.6,在低氧處理24 h時分彆為41.3±1.3、71.3±1.5、59.4±1.8,較常氧處理1.5 h(35.7±1.2、48.7±1.3、33.2±1.8)和24 h時(41.3±1.3、50.2±1.7、38.9±1.9)明顯增加,差異均有統計學意義(均P＜0.05).免疫組織化學結果也顯示低氧情況下胞質FAK、Grb2及吻蛋白的錶達增彊.結論 低氧狀態下HPASMC增殖與胞質FAK、Grb2及吻蛋白有關.這些胞質蛋白在低氧情況下對細胞生長和分化具有調控作用.
목적 탐토저양상태하점착반격매(FAK)대인폐동맥평활기세포(HPASMC)증식적영향궤제.방법 채용FAK과핵감산전염법대HPASMC진행저양처리,면역침정법측정포질단백FAK、생장인자수체결합단백2(Grb2)급문단백적활성,Western인적법측정포질단백FAK、Grb2급문단백적단백표체,면역조직화학법측정포질FAK、Grb2급문단백표체.결과 저양처리후포질FAK、Grb2급문단백적활성증가.포질FAK、Grb2급문단백적단백표체치재저양처리1.5 h시분별위43.4±1.4、69.7±1.9、59.3±1.6,재저양처리24 h시분별위41.3±1.3、71.3±1.5、59.4±1.8,교상양처리1.5 h(35.7±1.2、48.7±1.3、33.2±1.8)화24 h시(41.3±1.3、50.2±1.7、38.9±1.9)명현증가,차이균유통계학의의(균P＜0.05).면역조직화학결과야현시저양정황하포질FAK、Grb2급문단백적표체증강.결론 저양상태하HPASMC증식여포질FAK、Grb2급문단백유관.저사포질단백재저양정황하대세포생장화분화구유조공작용.
Objective To explore the mechanisms of focal adhesion kinase (FAK) in the proliferation of human pulmonary artery smooth muscle cells (HPASMCs) under hypoxia.Methods Cultured HPASMCs were passively transfected with FAK oligonucleotides (ODNS) and under normoxia or hypoxia condition.They were divided into four groups:normoxia without fibronectin ( FN), normoxia with FN, hypoxia without FN, hypoxia with FN in vitro respectively.Cytoplasmic FAK, Grb2 and paxillin were observed simultaneously by immunoprecipitation and Western blot.In addition, the expressions of cytoplasmic FAK, Grb2 and paxillin were detected by immunocytochemical staining.Results Immunoprecipitation and Western blot demonstrated that cytoplasmic expressions of FAK, Grb2 and paxillin in HPASMCs increased in hypoxia with FN from 43.4 ± 1.4, 69.7 ± 1.9, 59.3 ± 1.6 to 35.7 ± 1.2, 48.7±1.3, 33.2±1.8 at 1.5 h (all P＜0.05), from41.3±1.3, 71.3 ±1.5, 59.4 ±1.8 to41.3±1.3, 50.2 ± 1.7, 38.9 ± 1.9 at 24 h respectively ( P ＜ 0.01, P ＜ 0.05, P ＜ 0.05).Immunocytochemistry staining showed that the cytoplasmic expressions of FAK, Grb2 and paxillin were enhanced in hypoxia with FN versus normoxia with FN.There were significant differences.Conclusion Hypoxia can induce the activation of cytoplasmic FAK, Grb2 and paxillin so as to regulate the migration, survival and proliferation of HPASMCs.
