伏马毒素B1对人脐静脉血管内皮细胞的毒性作用研究

摘要

为探究伏马毒素B1(FB1)对人脐静脉血管内皮细胞(HUVEC)的毒性及作用机制,采用不同质量浓度的FB1处理HUVEC,通过CCK-8法检测细胞活力,流式细胞术检测细胞周期,荧光显微镜观察细胞氧化应激,流式细胞术检测线粒体膜电位变化,Hoechst33342染色检测细胞凋亡,荧光定量PCR检测细胞凋亡相关基因的转录情况.结果显示:FB1处理组在低质量浓度(2.5、5 μg· mL-1)短时间(6、12 h)处理后细胞活力变化不显著,超过此剂量和时间细胞活力显著降低(P＜0.05或P＜0.01),且质量浓度为10、20μg· mL-1 FB1处理组活性氧(ROS)的荧光信号增强;FB1处理组细胞周期由G0/G1期向S期转换时阻滞,细胞线粒体膜电位降低,细胞凋亡相关基因Cas pase-3转录升高,Bcl-2转录显著降低(P＜0.05),Caspase-9和Bax转录极显著升高(P＜0.01).结果表明,FB1对人脐静脉血管内皮细胞有毒性作用,能抑制细胞增殖,使细胞周期阻滞在G0/G1期;能通过线粒体途径诱导HUVEC凋亡.研究结果为深入研究FB1对人类细胞的毒性作用机制及相关疾病治疗奠定基础.%To investigate the toxicity and mechanism of action of fumonisin B1 (FB1) on human umbilical vein endothelial cells (HUVEC),HUVEC cells were treated with FB1 in different concentration.The cell vitality,cell cycle,cell oxidative stress,apoptosis,mitochondrial membrane potential change and transcription changes of apoptosis-related genes were detected by Cell Counting Kit-8,flow cytometry,fluorescence microscope,Hoechst33342 staining and real-time quantitative PCR.In result,the vitality of FB1 treated cells (FB1 dose exceed 5 μg · mL-1 and process time exceed 12 h) were significantly decrease (P＜0.01 or P＜0.05).And the fluorescence intensity of reactive oxygen active (ROS) were enhanced when being treated with 10,20μg · mL-1 of FB1.FB1 could prevent HUVEC from G0/G1 phase to S phase and reduce the mitochondrial membrane potential.The transcription of apoptosis gene Caspase-3 were increased,Bcl-2 were significantly declined (P＜0.05).Caspase-9 and Bax genes transcriptions were significantly increased (P＜0.01).The results showed that FB1 has cytotoxic effect on HUVEC cells,which could inhibit cell proliferation,arrest cell cycle at G0/G1 phase and induce apoptosis via mitochondrial pathway.Results in the present works provided the foundations for further research of toxicity and mechanism of FB1 on human cells and indications for some relevant diseases treatment.