摘要
为探究镉对牛肝成纤维细胞MT基因表达及细胞凋亡的影响,试验分别用不同浓度(0.625,1.25,2.5,5,10μmol/L)镉处理牛肝成纤维细胞24 h和48 h,观察细胞形态;采用实时荧光定量PCR方法检测各处理细胞中MT基因的相对表达量,并采用流式细胞术分别检测细胞的凋亡情况和细胞周期。结果表明:随着镉处理时间和浓度的增加,牛肝成纤维细胞的损伤逐渐加剧。处理24 h,与对照(未用镉处理的牛肝成纤维细胞)相比,1.25μmol/L镉处理的细胞中MT基因的相对表达量显著提高(P<0.05),2.5,5,10μmol/L镉处理的细胞中MT基因的相对表达量极显著提高(P<0.01),1.25μmol/L镉处理的细胞中MT基因的相对表达量显著提高(P<0.05),0.625μmol/L镉处理的细胞中MT基因的相对表达量差异不显著(P>0.05);处理48 h,与对照相比,0.625,1.25,2.5,5μmol/L镉处理的细胞中MT基因的相对表达量极显著提高(P<0.01),10μmol/L镉处理的细胞中MT基因的相对表达量差异不显著(P>0.05)。处理24 h,与对照相比,不同浓度镉处理的细胞凋亡率均差异不显著(P>0.05);处理48 h,2.5,5μmol/L镉处理的细胞凋亡率显著提高(P<0.05),10μmol/L镉处理的细胞凋亡率极显著提高(P<0.01),0.625,1.25μmol/L镉处理的细胞凋亡率差异不显著(P>0.05)。处理24 h,与对照相比,10μmol/L镉处理的细胞中G1期细胞占比极显著提高(P<0.01),0.625,1.25,2.5,5μmol/L镉处理的细胞中G1期细胞占比差异不显著(P>0.05);2.5μmol/L镉处理的细胞中S+G2期细胞占比显著降低(P<0.05),10μmol/L镉处理的细胞中S+G2期细胞占比极显著降低(P<0.01),0.625,1.25,5μmol/L镉处理的细胞中S+G2期细胞占比差异不显著(P>0.05)。处理48 h,与对照相比,0.625μmol/L镉处理的细胞中G1期细胞占比显著降低(P<0.05),5μmol/L镉处理的细胞中G1期细胞占比极显著提高(P<0.01),1.25,5,10μmol/L镉处理的细胞中G1期细胞占比差异不显著(P>0.05);0.625μmol/L镉处理的细胞中S+G2期细胞占比显著提高(P<0.05),5μmol/L镉处理的细胞中S+G2期细胞占比极显著降低(P<0.01),1.25,5,10μmol/L镉处理的细胞中S+G2期细胞占比差异不显著(P>0.05)。说明镉可导致牛肝成纤维细胞损伤,促进细胞中MT基因的表达,长时间(48 h)镉暴露会促进细胞凋亡,但长时间高浓度(2.5μmol/L以上)镉暴露会使MT基因表达量降低,同时高浓度镉会促进G1期细胞增殖,抑制S+G2期细胞增殖。
This study aimed to investigate the impacts of cadmium on MT gene expression and apoptosis in bovine liver fibroblasts.Bovine liver fibroblasts were exposed to varyous concentrations(0.625,1.25,2.5,5,10μmol/L)of cadmium for 24 h and 48 h respectively.Cell morphology was observed,and the relative expression levels of the MT gene in the treated cells were determined using real-time fluorescence quantitative PCR.Flow cytometry was employed to detect cell apoptosis and cell cycle.The results demonstrated that as the treatment time and cadmium concentration increased,the damage to bovine liver fibroblasts escalated.After 24 h of treatment,compared to the control(untreated cells),the relative expression level of the MT gene in bovine liver fibroblasts treated with 1.25μmol/L cadmium was significantly upregulated(P<0.05),and those treated with 2.5,5,10μmol/L cadmium showed an extremely significant upregulation(P<0.01)and those treated with 1.25μmol/L cadmium showed a sigmficant upregulation(P<0.05).There was no significant difference in the MT gene relative expression level between the 0.625μmol/L cadmium-treated cells(P>0.05).After 48 h of treatment,compared to the control,the MT gene relative expression levels in cells treated with 0.625,1.25,2.5 and 5μmol/L cadmium were extremely significantly elevated(P<0.01),while there was no significant difference in the 10μmol/L cadmium-treated cells(P>0.05).Regarding apoptosis,after 24 h of treatment,no significant difference was observed in the apoptosis rate of bovine liver fibroblasts treated with different cadmium concentrations compared to the control(P>0.05).After 48 h,the apoptosis rates of cells treated with 2.5 and 5μmol/L cadmium were significantly increased(P<0.05),and that of the 10μmol/L cadmium-treated cells was extremely significantly increased(P<0.01),with no significant difference in the 0.625 and 1.25μmol/L cadmium-treated cells(P>0.05).In terms of the cell cycle,after 24 h of treatment,compared to the control,the proportion of bovine liver fibroblasts in the G1 phase treated with 10μmol/L cadmium was extremely significantly increased(P<0.01),with no significant difference in the other concentrations.The proportion of cells in the S+G2 phase treated with 2.5μmol/L cadmium was significantly decreased(P<0.05),and that with 10μmol/L cadmium was extremely significantly decreased(P<0.01).After 48 h of treatment,compared to the control,the proportion of cells in the G1 phase treated with 0.625μmol/L cadmium was significantly decreased(P<0.05),and that with 5μmol/L cadmium was extremely significantly increased(P<0.01).The proportion of cells in the S+G2 phase treated with 0.625μmol/L cadmium was significantly increased(P<0.05),and that treared with 5μmol/L cadmium was extremely significantly decreased(P<0.01).Overall,cadmium could damage bovine liver fibroblasts,enhance the expression of the MT gene,and long-term(48 h)exposure of it would promote cell apoptosis,although long-term high-concentration(above 2.5μmol/L)exposure would reduce the MT gene expression.Additionally,high-concentration of cadmium would stimulate the proliferation of cells in the G1 phase and suppress the proliferation of cells in the S+G2 phase.
作者
张凯凯
赵元峰
胡荣斌
冉江
代国滔
ZHANG Kaikai;ZHAO Yuanfeng;HU Rongbin;RAN Jiang;DAI Guotao(Guizhou Animal Husbandry and Veterinary Research Institute,Guiyang 550005,China;Sinan County Agricultural and Rural Bureau,Tongren 565100,China)
出处
《黑龙江畜牧兽医》
北大核心
2025年第3期13-20,共8页
Heilongjiang Animal Science And veterinary Medicine
基金
贵州省科技计划项目(黔科合基础-ZK[2022]一般226)
贵州省农业科学院青年基金项目(黔农科青年基金[2023]04号)
贵州省肉牛现代农业产业技术体系建设项目(GZRNCYJSTX-02)。
