中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
Chinese Journal of Nephrology
2015年
9期
680-685
,共6页
棕榈酸%足细胞%细胞凋亡%转录激活因子类
棕櫚痠%足細胞%細胞凋亡%轉錄激活因子類
종려산%족세포%세포조망%전록격활인자류
Palmitic acid%Podocytes%Apoptosis%Activating transcription factors
目的 探讨转录激活因子6(activating transcription factor 6 ,ATF6)在棕榈酸(palmitic acid ,PA)诱导的足细胞(MPC5细胞)凋亡和增殖中的作用. 方法 不同浓度PA刺激足细胞24 h ,检测ATF6和激活型天冬氨酸特异性半胱氨酸蛋白酶(Cleaved?caspase3)表达水平及足细胞凋亡情况.腺病毒siATF6转染沉默ATF6后,比较PA诱导的足细胞增殖能力、细胞周期、细胞凋亡和Cleaved?caspase3的蛋白表达水平的变化.采用Western印迹检测Cleaved?caspase3和ATF6蛋白表达水平,流式细胞术检测足细胞凋亡和细胞周期,甲基噻唑基四唑(MTT)法检测细胞增殖,细胞免疫荧光观察ATF6在足细胞中的表达和分布. 结果 与对照组比较,PA诱导组足细胞凋亡明显增加(P<0.05),细胞Cleaved?caspase3和ATF6蛋白水平随PA浓度增加呈剂量依赖性增高(均P<0.05).采用腺病毒siATF6转染足细胞后,PA诱导的足细胞凋亡较空载体转染组减少,细胞增殖增加,S期的细胞比例明显增多,Cleaved?caspase3的表达降低(均P<0.05);空载体转染组与PA诱导组在上述指标间的差异均无统计学意义. 结论 ATF6介导棕榈酸诱导的足细胞凋亡.
目的 探討轉錄激活因子6(activating transcription factor 6 ,ATF6)在棕櫚痠(palmitic acid ,PA)誘導的足細胞(MPC5細胞)凋亡和增殖中的作用. 方法 不同濃度PA刺激足細胞24 h ,檢測ATF6和激活型天鼕氨痠特異性半胱氨痠蛋白酶(Cleaved?caspase3)錶達水平及足細胞凋亡情況.腺病毒siATF6轉染沉默ATF6後,比較PA誘導的足細胞增殖能力、細胞週期、細胞凋亡和Cleaved?caspase3的蛋白錶達水平的變化.採用Western印跡檢測Cleaved?caspase3和ATF6蛋白錶達水平,流式細胞術檢測足細胞凋亡和細胞週期,甲基噻唑基四唑(MTT)法檢測細胞增殖,細胞免疫熒光觀察ATF6在足細胞中的錶達和分佈. 結果 與對照組比較,PA誘導組足細胞凋亡明顯增加(P<0.05),細胞Cleaved?caspase3和ATF6蛋白水平隨PA濃度增加呈劑量依賴性增高(均P<0.05).採用腺病毒siATF6轉染足細胞後,PA誘導的足細胞凋亡較空載體轉染組減少,細胞增殖增加,S期的細胞比例明顯增多,Cleaved?caspase3的錶達降低(均P<0.05);空載體轉染組與PA誘導組在上述指標間的差異均無統計學意義. 結論 ATF6介導棕櫚痠誘導的足細胞凋亡.
목적 탐토전록격활인자6(activating transcription factor 6 ,ATF6)재종려산(palmitic acid ,PA)유도적족세포(MPC5세포)조망화증식중적작용. 방법 불동농도PA자격족세포24 h ,검측ATF6화격활형천동안산특이성반광안산단백매(Cleaved?caspase3)표체수평급족세포조망정황.선병독siATF6전염침묵ATF6후,비교PA유도적족세포증식능력、세포주기、세포조망화Cleaved?caspase3적단백표체수평적변화.채용Western인적검측Cleaved?caspase3화ATF6단백표체수평,류식세포술검측족세포조망화세포주기,갑기새서기사서(MTT)법검측세포증식,세포면역형광관찰ATF6재족세포중적표체화분포. 결과 여대조조비교,PA유도조족세포조망명현증가(P<0.05),세포Cleaved?caspase3화ATF6단백수평수PA농도증가정제량의뢰성증고(균P<0.05).채용선병독siATF6전염족세포후,PA유도적족세포조망교공재체전염조감소,세포증식증가,S기적세포비례명현증다,Cleaved?caspase3적표체강저(균P<0.05);공재체전염조여PA유도조재상술지표간적차이균무통계학의의. 결론 ATF6개도종려산유도적족세포조망.
Objective To observe the effect of ATF6 on the apoptosis and proliferation of podocytes induced by palmitic acid (PA). Methods Podocytes were stimulated with different doses of PA for 24 h. The expression of cleaved-caspase3 was detected by Western blotting. The podocyte apoptosis was analyzed by flow cytometry (FCM), and the expression of ATF6 was tested by Western blotting and immunofluorescence staining. After the transfection of adenovirus siRNA against ATF6, the proliferation, the cell cycle and apoptosis of potocytes stimulated with PA were tested by MTT or FCM. Results The levels of cleaved - caspase3 and ATF6 of podocytes stimulated with PA were significantly increased by a dose-dependent manner compared with the control group (P<0.05). The apoptosis of podocytes stimulated with PA was increased (P<0.05). Compared with the podocytes stimulated with PA, the apoptosis of podocytes transfected by adenovirus siRNA against ATF6 with PA stimulation was significantly reduced (P<0.05). The proliferation of podocytes transfected by adenovirus siRNA against ATF6 and stimulated with PA, however, was obviously increased compared with the podocytes stimulated with PA (P<0.05). Conclusion ATF6 mediated the apoptosis of podocytes induced by palmitate acid.
