锰对人脐静脉内皮细胞及半胱天冬酶表达影响
Effect of manganese on growth of HUVEC-304 cells and expression of caspase
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摘要: 目的探讨锰对人脐静脉内皮细胞系HUVEC-304细胞生长周期及半胱天冬酶-3(caspas-3)、半胱天冬酶-8(caspase-8)表达的影响。方法以100~800μmoml/L)的氯化锰分别处理HUVEC-304细胞24~72 h后,四甲基偶氮噻唑蓝(MTT)法检测EVC-304细胞的生长活性;流式细胞仪(FCM)检测细胞凋亡情况;蛋白印迹法(Westernblot)检测HUVEC-304细胞在,氯化锰半数抑制浓度(IC50)400μmol/L作用24 h时caspase-8、caspase-3的表达。结果氯化锰可呈时间及剂量依赖性抑制HUVEC-304细胞的增殖和诱导细胞凋亡,抑制率为22.34%~90.94%,凋亡率为10.20%~98.73%。氯化锰24 hIC50约为400μmol/L,在此浓度下,HUVEC-304细胞caspase-8、caspase-3表达显著增高,(P<0.05)。结论氯化锰能够抑制HUVEC-304细胞的增殖,呈明显的时效和量效关系。caspase-8、cas-pase-3表达增高在细胞增殖和凋亡中起重要的作用,可能是其作用机制之一。Abstract: ObjectiveTo investigate the effect of manganese on growth of HUVEC-304 cells and the ex pression of caspase-8、caspase-3.MethodHUVEC 304 cells were given with various concentrations of MnCl2.The inhibitory ratio was measured by MTT assay.Apoptosis was observed by flow cytometry(FCM).The expression of caspase-3 and caspase-8 was measured by Western blot.ResultsMTT results revealed that manganese chloride(from 100 to 800 μmol/L)might suppress the proliferation of HUVEC-304 cells(inhibitory ratio were 22.34%~90.94%)and induce apoptosis(apoptosis ratio were 10.20%~98.73%)in dose and time-dependent manner.The result of Western Blot showed that manganese inhibited caspase-3 and caspase-8 expression(P<0.05).ConclusionManganese can inhibit proliferation of HUVEC-304 cells with time-and dose-dependent manner.These may be one of important mechanisms that manganese suppressed the proliferation of HUVEC-304 cells and induced apoptosis.
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