中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2015年
8期
598-603
,共6页
沈玥%严玉澄%路丽明%钱盈盈%关雪晶%倪兆慧%钱家麒
瀋玥%嚴玉澄%路麗明%錢盈盈%關雪晶%倪兆慧%錢傢麒
침모%엄옥징%로려명%전영영%관설정%예조혜%전가기
氧化性应激%细胞凋亡%丝裂原激活蛋白激酶%Klotho
氧化性應激%細胞凋亡%絲裂原激活蛋白激酶%Klotho
양화성응격%세포조망%사렬원격활단백격매%Klotho
Oxidative stress%Apoptosis%Mitogen-activated protein kinase%Klotho
目的 研究过氧化氢(H2O2)引起的氧化应激损伤对小鼠肾小管上皮细胞(TCMK-1)Klotho蛋白表达的影响及可能的作用机制.方法 体外培养TCMK-1细胞,以不同浓度的H2O2刺激细胞,采用流式细胞术检测活性氧族(ROS)生成;CCK-8检测细胞生存率;流式细胞术和Hoechst 33258染色检测细胞凋亡;Western印迹法检测Klotho蛋白、细胞凋亡相关蛋白和抗氧化酶的表达.结果 与正常对照组比较,H2O2刺激后TCMK-1细胞内ROS水平升高(均P＜ 0.05),抗氧化酶锰超氧化物歧化酶(manganese superoxide dismutase,SOD2)和过氧化氢酶(catalase,CAT)表达水平降低(均P＜0.05);Klotho蛋白的表达水平降低(均P＜0.05);TCMK-1细胞生存率显著降低(均P＜0.05),且呈剂量依赖(0.3～0.9 mmol/L H2O2);TCMK-1凋亡细胞比例呈剂量依赖性增加(均P＜ 0.05);Bax/Bcl-2表达比升高,JNK、p38蛋白磷酸化水平均升高(均P＜ 0.05).结论 H2O2刺激TCMK-1细胞诱导氧化应激损伤,抑制Klotho蛋白的表达,而Klotho表达下降伴随着细胞凋亡的增加和p38、JNK通路的活化.
目的 研究過氧化氫(H2O2)引起的氧化應激損傷對小鼠腎小管上皮細胞(TCMK-1)Klotho蛋白錶達的影響及可能的作用機製.方法 體外培養TCMK-1細胞,以不同濃度的H2O2刺激細胞,採用流式細胞術檢測活性氧族(ROS)生成;CCK-8檢測細胞生存率;流式細胞術和Hoechst 33258染色檢測細胞凋亡;Western印跡法檢測Klotho蛋白、細胞凋亡相關蛋白和抗氧化酶的錶達.結果 與正常對照組比較,H2O2刺激後TCMK-1細胞內ROS水平升高(均P＜ 0.05),抗氧化酶錳超氧化物歧化酶(manganese superoxide dismutase,SOD2)和過氧化氫酶(catalase,CAT)錶達水平降低(均P＜0.05);Klotho蛋白的錶達水平降低(均P＜0.05);TCMK-1細胞生存率顯著降低(均P＜0.05),且呈劑量依賴(0.3～0.9 mmol/L H2O2);TCMK-1凋亡細胞比例呈劑量依賴性增加(均P＜ 0.05);Bax/Bcl-2錶達比升高,JNK、p38蛋白燐痠化水平均升高(均P＜ 0.05).結論 H2O2刺激TCMK-1細胞誘導氧化應激損傷,抑製Klotho蛋白的錶達,而Klotho錶達下降伴隨著細胞凋亡的增加和p38、JNK通路的活化.
목적 연구과양화경(H2O2)인기적양화응격손상대소서신소관상피세포(TCMK-1)Klotho단백표체적영향급가능적작용궤제.방법 체외배양TCMK-1세포,이불동농도적H2O2자격세포,채용류식세포술검측활성양족(ROS)생성;CCK-8검측세포생존솔;류식세포술화Hoechst 33258염색검측세포조망;Western인적법검측Klotho단백、세포조망상관단백화항양화매적표체.결과 여정상대조조비교,H2O2자격후TCMK-1세포내ROS수평승고(균P＜ 0.05),항양화매맹초양화물기화매(manganese superoxide dismutase,SOD2)화과양화경매(catalase,CAT)표체수평강저(균P＜0.05);Klotho단백적표체수평강저(균P＜0.05);TCMK-1세포생존솔현저강저(균P＜0.05),차정제량의뢰(0.3～0.9 mmol/L H2O2);TCMK-1조망세포비례정제량의뢰성증가(균P＜ 0.05);Bax/Bcl-2표체비승고,JNK、p38단백린산화수평균승고(균P＜ 0.05).결론 H2O2자격TCMK-1세포유도양화응격손상,억제Klotho단백적표체,이Klotho표체하강반수착세포조망적증가화p38、JNK통로적활화.
Objective To evaluate the effect of oxidative injury induced by peroxide oxidase on Klotho expression in mouse renal tubular epithelial cells (TCMK-1) and to explore the possible pathway.Methods TCMK-1 cells were exposed to H2O2 of different concentrations.Reactive oxygen species (ROS) was examined byflow cytometrry.Cell viability was assessed by CCK-8.Cell apoptosis was evaluated by flow cytometry and Hoechst 33258 staining.The expression of Klotho,apoptosis-associated proteins and anti-oxidant enzymes were determined by Western blotting.Results Compared with control group,after H2O2 stimulating TCMK-1 cell,ROS was dramatically elevated (all P ＜ 0.05) and the expression of anti-oxidant enzymes,SOD2 and CAT went down (all P ＜ 0.05);the expression of Klotho was inhibited (all P ＜ 0.05);cell viability of TCMK-1 cells was decreased (all P ＜ 0.05) in a dose-dependent manner (0.3 to 0.9 mmol/L);cell apoptosis was significantly increased in TCMK-1 cells following the concentration of H2O2 (all P ＜ 0.05);Bax/Bcl-2 and the phosphororation of JNK and p38 were obviously elevated in TCMK-1 by H2O2 induction (all P ＜ 0.05).Conclusion Oxidative injuries induced by H2O2 significantly suppresses the expression of Klotho in TCMK-1 cells.And cell apoptosis was increased,p38 and JNK pathway was activated.
