中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2015年
5期
370-373
,共4页
阮滔%何学华%刘丽萍%袁勇华%罗建红%潘丽%胡沙雅
阮滔%何學華%劉麗萍%袁勇華%囉建紅%潘麗%鬍沙雅
원도%하학화%류려평%원용화%라건홍%반려%호사아
Wnt2%β-catenin%多柔比星%心肌损伤%p53
Wnt2%β-catenin%多柔比星%心肌損傷%p53
Wnt2%β-catenin%다유비성%심기손상%p53
Wnt2%β-catenin%Doxorubicin%Myocardial injury%p53
目的 研究Wnt2、β-catenin蛋白在多柔比星(DOX)诱导心肌损伤中的表达情况,探讨Wnt2、β-catenin分子在DOX损伤心肌细胞凋亡中的作用.方法 将不同质量浓度(1 mg/L、2 mg/L、3 mg/L、4 mg/L)DOX处理心肌细胞72 h后,用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测不同质量浓度DOX对心肌细胞生长曲线的影响.用1 mg/L DOX建立心肌细胞损伤模型,将心肌细胞分为4组,A组:DOX损伤心肌细胞12 h;B组:DOX损伤心肌细胞24 h;C组:DOX损伤心肌细胞48 h;D组:正常心肌细胞.利用蛋白印迹法(Western blot)、实时反转录聚合酶链反应(RT-PCR)法分别检测各组细胞中Wnt2、β-catenin、p53蛋白及mRNA的表达变化.结果 DOX抑制对心肌细胞的增殖具有浓度依赖性;Wnt2蛋白和mRNA的表达在DOX各组中表达水平均高于D组,差异有统计学意义(F =224.115,P＜0.05);β-catenin、p53蛋白在DOX各组中的表达水平显著高于D组,且随时间的延长,表达程度越高,差异均有统计学意义(F=188.145、231.927,P均＜0.05);β-catenin与p53呈正相关(r=0.940,P＜0.05).结论 Wnt2、β-catenin可能在DOX诱导的心肌细胞损伤中起着重要的作用,为进一步探讨心肌细胞损伤的分子机制提供了重要依据.
目的 研究Wnt2、β-catenin蛋白在多柔比星(DOX)誘導心肌損傷中的錶達情況,探討Wnt2、β-catenin分子在DOX損傷心肌細胞凋亡中的作用.方法 將不同質量濃度(1 mg/L、2 mg/L、3 mg/L、4 mg/L)DOX處理心肌細胞72 h後,用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴鹽(MTT)法檢測不同質量濃度DOX對心肌細胞生長麯線的影響.用1 mg/L DOX建立心肌細胞損傷模型,將心肌細胞分為4組,A組:DOX損傷心肌細胞12 h;B組:DOX損傷心肌細胞24 h;C組:DOX損傷心肌細胞48 h;D組:正常心肌細胞.利用蛋白印跡法(Western blot)、實時反轉錄聚閤酶鏈反應(RT-PCR)法分彆檢測各組細胞中Wnt2、β-catenin、p53蛋白及mRNA的錶達變化.結果 DOX抑製對心肌細胞的增殖具有濃度依賴性;Wnt2蛋白和mRNA的錶達在DOX各組中錶達水平均高于D組,差異有統計學意義(F =224.115,P＜0.05);β-catenin、p53蛋白在DOX各組中的錶達水平顯著高于D組,且隨時間的延長,錶達程度越高,差異均有統計學意義(F=188.145、231.927,P均＜0.05);β-catenin與p53呈正相關(r=0.940,P＜0.05).結論 Wnt2、β-catenin可能在DOX誘導的心肌細胞損傷中起著重要的作用,為進一步探討心肌細胞損傷的分子機製提供瞭重要依據.
목적 연구Wnt2、β-catenin단백재다유비성(DOX)유도심기손상중적표체정황,탐토Wnt2、β-catenin분자재DOX손상심기세포조망중적작용.방법 장불동질량농도(1 mg/L、2 mg/L、3 mg/L、4 mg/L)DOX처리심기세포72 h후,용3-(4,5-이갑기새서-2)-2,5-이분기사담서추염(MTT)법검측불동질량농도DOX대심기세포생장곡선적영향.용1 mg/L DOX건립심기세포손상모형,장심기세포분위4조,A조:DOX손상심기세포12 h;B조:DOX손상심기세포24 h;C조:DOX손상심기세포48 h;D조:정상심기세포.이용단백인적법(Western blot)、실시반전록취합매련반응(RT-PCR)법분별검측각조세포중Wnt2、β-catenin、p53단백급mRNA적표체변화.결과 DOX억제대심기세포적증식구유농도의뢰성;Wnt2단백화mRNA적표체재DOX각조중표체수평균고우D조,차이유통계학의의(F =224.115,P＜0.05);β-catenin、p53단백재DOX각조중적표체수평현저고우D조,차수시간적연장,표체정도월고,차이균유통계학의의(F=188.145、231.927,P균＜0.05);β-catenin여p53정정상관(r=0.940,P＜0.05).결론 Wnt2、β-catenin가능재DOX유도적심기세포손상중기착중요적작용,위진일보탐토심기세포손상적분자궤제제공료중요의거.
Objective To investigate the expressions of Wnt2 and β-catenin in Doxorubicin (DOX)-induced myocardial injury and to explore their roles in myocardial cell apoptosis.Methods Cardiomyoblast cells were damaged by different concentrations of DOX(1 mg/L,2 mg/L,3 mg/L,4 mg/L) for 72 h.The effect of different concentrations of DOX on cardiomyocyte growth curve was detected according to the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-h-tetrazolium bromide(MTT) assay.DOX(1 mg/L) was used to induce the model of cardiomyoblast cell injury.Cardiomyocytes were divided into 4 groups:group A:DOX-injured cardiomyocytes for 12 h ;group B:DOX-injured cardiomyocytes for 24 h ; group C:DOX-injured cardiomyocytes for 48 h; group D:normal cardiomyocytes.The expressions of Wnt2,β-catenin and p53 were observed by Western blot and reverse transcription polymerase chain reaction(RT-PCR) at the time point of 12 h,24 h and 48 h.Results DOX significantly inhibited cardiomyocyte proliferation in a dose dependent fashion.The protein and mRNA expressions of Wnt2 increased in the DOX-induced myocardial injury group compared with the group D,with statistical significance (F =224.115,P ＜ 0.05) ;The expressions of β-catenin,p53 were significantly increased compared with the group D,and the higher expression appeared with the time extending(F =188.145,231.927,all P ＜ 0.05).Significantly positive correlation between Wnt2 and β-catenin expression was observed(r =0.940,P ＜ 0.05).Conclusions These findings suggest that Wnt2/β-catenin signaling pathway may play important roles in the cardiovascular disease and be useful for exploring the molecular mechanism of myocardial injury..
