线粒体调控在玉米赤霉烯酮与脱氧雪腐镰刀菌烯醇联合染毒诱导仔猪睾丸支持细胞内质网途径凋亡中的作用  

Effects of Mitochondrial Regulation on Endoplasmic Reticulum Pathway Apoptosis of Sertoli Cells Induced by Zearalenone and Deoxynivalenol in Piglets

作　　者：陈佳雯 海思娆 马立 裘知 韩梦怡 王晨龙 冯士彬 赵畅 王希春[1] CHEN Jiawen;HAI Sirao;MA Li;QIU Zhi;HAN Mengyi;WANG Chenlong;FENG Shibin;ZHAO Chang;WANG Xichun(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China)

机构地区：[1]安徽农业大学动物科技学院,合肥230036

出　　处：《动物营养学报》2023年第2期1261-1269,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：安徽省自然科学基金项目(2208085MC81);安徽省大学生创新创业训练计划项目(S202110364260)。

摘　　要：本试验旨在研究线粒体在玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合染毒诱导仔猪睾丸支持细胞(SCs)内质网途径凋亡中的调控作用机制。将对数生长期仔猪SCs分为对照、ZEA+DON、线粒体分裂抑制因子-1(Mdivi-1)和ZEA+DON+Mdivi-14个处理,处理24 h后,采用荧光显微镜观察细胞存活率,采用透射电镜检测细胞超微结构、线粒体及内质网形态变化,采用流式细胞术检测细胞凋亡率,采用实时荧光定量PCR(qRT-PCR)检测细胞内质网途径凋亡相关基因蛋白激酶R样内质网激酶(PERK)、活化转录因子4(ATF4)、CCAAT/增强子结合蛋白同源蛋白(CHOP)和真核生物翻译起始因子2α(eIF2α)mRNA相对表达量,并采用蛋白质印迹法(Western blot)检测内质网途径凋亡相关蛋白PERK、ATP4、CHOP和eIF2α蛋白相对表达量。结果表明:1)与ZEA+DON处理相比,添加Mdivi-1可极显著提高细胞存活率(P<0.01),缓解细胞、线粒体及内质网结构损伤。2)与对照处理相比,Mdivi-1处理细胞凋亡率无显著差异(P>0.05),而ZEA+DON处理和ZEA+DON+Mdivi-1处理细胞凋亡率极显著提高(P<0.01);但与ZEA+DON处理相比,ZEA+DON+Mdivi-1处理细胞凋亡率极显著降低(P<0.01)。3)与对照处理相比,ZEA+DON处理和ZEA+DON+Mdivi-1处理PERK、ATF4、CHOP及eIF2αmRNA相对表达量和蛋白相对表达量极显著提高(P<0.01);但与ZEA+DON处理相比,ZEA+DON+Mdivi-1处理PERK、ATF4、CHOP、eIF2αmRNA相对表达量和蛋白相对表达量显著或极显著降低(P<0.05或P<0.01)。由此可知,线粒体通过内质网途径在ZEA与DON联合染毒仔猪SCs凋亡中发挥重要作用,干预该过程可为霉菌毒素的生殖毒性危害解除提供新思路。This experiment was conducted to investigate the regulatory mechanism of mitochondria on endoplasmic reticulum pathway apoptosis of Sertoli cells(SCs)induced by combined treatment with zearalenone(ZEA)and deoxynivalenol(DON)in piglets.The SCs at logarithmic growth stage were divided into control,ZEA+DON,mitochondrial division inhibitor-1(Mdivi-1)and ZEA+DON+Mdivi-1 treatments.After 24 h of treatment,the cell viability was observed by fluorescence microscope;the ultrastructure,mitochondria and endoplasmic reticulum morphology were detected by transmission electron microscope;the cell apoptosis rate was detected by flow cytometry;the mRNA relative expression levels of protein kinase R-like endoplasmic reticulum kinase(PERK),activating transcription factor 4(ATF4),CCAAT/enhancer binding protein homolog(CHOP)and eukaryotic translation initiation factor 2α(eIF2α)genes in endoplasmic reticulum apoptosis pathway were detected by real-time quantitative PCR;and the protein relative expression levels of PERK,ATP4,CHOP and eIF2αwere detected by Western blot.The results showed as follows:1)compared with ZEA+DON treatment,adding Mdivi-1 extremely significantly increased the cell viability(P<0.01),and alleviated the structural damage of cells,mitochondria and endoplasmic reticulum.2)Compared with the control treatment,the cell apoptosis rate in Mdivi-1 treatment had no significant difference(P>0.05),while the cell apoptosis rate in ZEA+DON and ZEA+DON+Mdivi-1 treatments were extremely significantly increased(P<0.01);compared with ZEA+DON treatment,the cell apoptosis rate in ZEA+DON+Mdivi-1 treatment was extremely significantly decreased(P<0.01).3)Compared with the control treatment,the mRNA relative expression levels and protein relative expression levels of PERK,ATF4,CHOP and eIF2αin ZEA+DON and ZEA+DON+Mdivi-1 treatments were extremely significantly increased(P<0.01);but compared with ZEA+DON treatment,the mRNA relative expression levels and protein relative expression levels of PERK,ATF4,CHOP and eIF2αin ZEA+DON+Mdivi-

关 键 词：玉米赤霉烯酮 脱氧雪腐镰刀菌烯醇 仔猪睾丸支持细胞 线粒体 内质网途径凋亡 

分 类 号：S811[农业科学—畜牧学]