摘要
为了探讨乙酰左旋肉碱(acetyl L-carnitine, ALC)对高锌(zinc, Zn)诱导PK-15细胞凋亡的影响,本研究首先通过CCK-8法检测不同浓度的Zn和ALC对细胞活力的影响,并选择最佳的处理浓度进行24 h细胞干预。细胞试验分为对照组、Zn组、Zn+ALC组和ALC组。试验采用实时荧光定量PCR检测细胞凋亡相关基因bax、bak-1、bcl-2和Caspase-3的mRNA转录水平,免疫印迹检测Caspase-3和cleaved Caspase-3的蛋白表达。结果显示,100 mmol/L的ALC可以显著提高PK-15的细胞活力(P<0.05),Zn在PK-15细胞活力中的半数抑制浓度为121.945μmol/L。Zn处理细胞可显著上调凋亡相关基因bax、bak-1和Caspase-3的mRNA表达以及cleaved Caspase-3的蛋白表达(P<0.05)。与Zn处理组相比,Zn+ALC组中bax、bak-1和Caspase-3的mRNA表达以及cleaved Caspase-3的蛋白表达呈现显著降低(P<0.05)。此外,Zn组中bcl-2的mRNA表达和Caspase-3蛋白表达显著低于对照组(P<0.05),而ALC与Zn联合作用可逆转这一效应(P<0.05)。结果表明,ALC可有效缓解Zn诱导PK-15细胞的凋亡效应,对高Zn导致的毒性损伤具有一定的保护作用。
In order to investigate the effect of acetyl L-carnitine(ALC) on the apoptosis induced by high level of zinc(Zn) in PK-15 cells, the effects of different concentrations of Znand ALC on cell viability were detected by CCK-8 method, and the optimal treatment concentration was selected for 24 h-intervention.In this study, the cells were divided into control group, Zn group, Zn+ALC group and ALC group.Meanwhile, the mRNA expression levels of apoptosis-related genes(bax, bak-1, bcl-2 and Caspase-3) were detected by real-time quantitative PCR,and protein expression levels of Caspase-3 and cleaved Caspase-3 were determined by Western blot.The results showed that 100 mmol/L ALC could significantly increase the cell viability of PK-15 cells(P<0.05).The 50% inhibitory concentration of Znwas 121.945 μmol/L.Subsequently, the mRNA expression levels of apoptosis-related genes(bax,bak-1 and Caspase-3) and the protein expression level of cleaved Caspase-3 were markedly increased under Zntreatment(P<0.05).Furthermore, compared to the Zn group, the mRNA expression of apoptosis-related genes(bax,bak-1 and Caspase-3) and the protein expression level of cleaved Caspase-3 in Zn+ALC group were significantly decreased(P<0.05).In addition,the mRNA expression of bcl-2and protein level of Caspase-3were remarkably decreased in Zn group,while ALC combined with Zn could reverse this effect(P<0.05).These results suggested that ALC could effectively alleviate the Zn-induced apoptosis in PK-15cells and protect against the toxic damage caused by high levels of Zn.
作者
杨庆稳
雍康
张怡
贺闪闪
杨钧杰
彭津津
聂青玉
吴有华
YANG Qingwen;YONG Kang;ZHANG Yi;HE Shanshan;YANG Junjie;PENG Jinjin;NIE Qingyu;WU Youhua(Chongqing Three Gorges Vocational College,Wanzhou,Chongqing 404155,China;College of Agriculture,Guangdong Ocean University,Zhanjiang,Guangdong 524088,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2022年第10期2059-2063,共5页
Chinese Journal of Veterinary Science
基金
重庆市自然科学基金资助项目(cstc2021jcyj-msxmX1210)
重庆市教委科研课题基金资助项目(KJQN201903503)
重庆市教委教改课题基金资助项目(Z213122)
三峡库区特色农产品质量与安全基金资助项目(CXQTP19037)。
