文章摘要
壳寡糖对黄曲霉毒素B1诱导大鼠肝细胞毒性损伤的干预作用
Intervention Effect of Chitooligosaccharides on Aflatoxin B1 Induced Toxic Damage of Rat Liver Cells
投稿时间:2020-11-24  修订日期:2021-01-04
DOI:
中文关键词: 黄曲霉毒素B1、壳寡糖、细胞毒性、抗氧化
英文关键词: Aflatoxin  B1, Chitooligosaccharide, Cytotoxicity, Antioxidant
基金项目:国家重点研发计划(2018YFC1602205)
作者单位邮编
谢佳雨 上海海洋大学 201306
张雯 上海海洋大学 食品学院 201306
杨靖亚 上海海洋大学 食品学院 201306
严佳惠 上海海洋大学 食品学院 201306
欧杰 上海海洋大学 食品学院 201306
摘要点击次数: 83
全文下载次数: 0
中文摘要:
      目的:研究壳寡糖(COS)对黄曲霉毒素B1(AFB1)诱导大鼠肝细胞(BRL 3A细胞)毒性损伤的干预作用。方法:采用CCK-8法分别测定AFB1对细胞的IC50和COS对细胞的无损害作用浓度;用试剂盒检测COS预处理细胞6 h后再加入AFB1继续培养24 h的细胞存活率、活性氧(ROS)水平、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、谷胱甘肽S-转移酶(GST)活性和细胞凋亡率;通过Real-time quantitative PCR(RT-qPCR)测定Nrf2、Keap1、Ho-1、Nqo1、Bax和Bcl-2基因的mRNA相对表达量;最后通过RNA-seq研究分析差异表达基因的层次聚类和富集途径。结果:AFB1对BRL 3A细胞的IC50为15.86 μmol/L,COS浓度小于125 μmol/L时不会对细胞造成毒性损伤;COS可以缓解AFB1引起的细胞内ROS水平和MDA含量升高,增强SOD和GST酶活性,进而提高细胞自身的抗氧化能力,降低细胞凋亡率;AFB1可引起促凋亡基因Bax的显著表达(P<0.05),并显著降低Nrf2、Keap1、Ho-1、Nqo1的转录水平(P<0.05),而COS预处理能显著提高Nrf2、Keap1、Ho-1、Nqo1基因的表达(P<0.05),显著降低Bax的表达水平(P<0.05);RNA-seq的富集途径结果中,COS还可能通过细胞色素P450对外源物质的代谢作用、药物代谢-细胞色素P450和p53信号通路途径来缓解AFB1诱导的细胞毒性损伤(P<0.05)。结论:COS预处理对AFB1诱导大鼠肝细胞的毒性损伤具有干预作用,其机制可能与Nrf2信号通路、细胞色素P450对外源物质的代谢作用、药物代谢-细胞色素P450和p53信号通路有关。
英文摘要:
      Objective: To study the effect of chitooligosaccharides(COS) on aflatoxin B1(AFB1)-induced toxic injury of rat liver cells(BRL 3A cells). Methods: The CCK-8 method was used to determine the IC50 value of AFB1 on cells and the non-damage concentration of COS on cells. After COS pretreated the cells for 6 hours and added AFB1 to continue culturing for 24 hours, we used the kit to determine the cell viability, reactive oxygen species (ROS) levels, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, glutathione S-transferase (GST) activity and apoptosis rate. The mRNA relative expression levels of Nrf2, Keap1, Ho-1, Nqo1, Bax and Bcl-2 genes were determined by real-time quantitative PCR (RT qPCR), and the hierarchical clustering and enrichment pathways of differentially expressed genes were analyzed by RNA-seq. Results: The IC50 of AFB1 to BRL 3A cells was 15.86 μmol/L, and the COS concentration less than 125 μmol/L would not cause toxic damage to the BRL 3A cells. COS can reduce the increase of ROS level and MDA content in cells induced by AFB1, enhance the activity of SOD and GST, thereby improve the cell"s own antioxidant capacityand reduce the rate of apoptosis. AFB1 can cause the significant expression of the pro-apoptotic gene Bax (P<0.05), and significantly reduce the transcription levels of Nrf2, Keap1, Ho-1, and Nqo1 (P<0.05), while COS pretreatment can significantly increase the expression of Nrf2, Keap1, Ho-1, Nqo1(P<0.05), and significantly reducing the expression level of Bax (P<0.05). In the results of RNA-seq enrichment pathways, COS may also alleviate the cytotoxic damage induced by AFB1 through the metabolism of xenobiotics by cytochrome P450, drug metabolism-cytochrome P450 and p53 signaling pathway (P<0.05). Conclusion: COS pretreatment can intervene the toxic injury of BRL 3A cells induced by AFB1, which may be mediated by Nrf2 signaling pathway, metabolism of xenobiotics by cytochrome P450, drug metabolism-cytochrome P450 and p53 signaling pathway.
HTML View Fulltext     下载PDF阅读器
关闭