动物营养学报2019，Vol.31Issue(11)：5379-5391,13.DOI:10.3969/j.issn.1006⁃267x.2019.11.055
慢性铅胁迫对日本沼虾硫氧还蛋白和热应激蛋白系统基因表达及肠道菌群的影响
Effects of Chronic Lead Exposure on Expressions of Thioredoxin and Heat Shock Protein System Genes and Intestinal Microbiota in Macrobrachium nipponense
摘要
Abstract
The aim of this study was to investigate the effects of chronic lead exposure on expressions of thiore?doxin and heat shock protein system genes and intestine microbiota in Macrobrachium nipponense. Juvenile M. nipponense with an average weight of (0.10±0.02) g were randomly divided into three groups in three repli?cates (50 tails／replicate) for chronic lead exposure trial. The concentration of lead in water were 0 ( control group) , 13.13 and 26.26 μg／L and exposure time lasted for 60 days. After the exposure trial, quantitative re?al?time polymerase chain reaction ( qRT?PCR ) was used to analyze the mRNA expressions of thioredoxin ( Trx) , thioredoxin reductase ( TrxR ) , heat shock protein 60 ( HSP60 ) , heat shock cognate protein 70 ( HSC70) and heat shock protein 90 ( HSP90). Meanwhile, the composition and diversity of intestine microbi?ota were analyzed by 16s RNA high throughput sequencing. The results showed as follows 1) chronic lead ex?posure inhibited the mRNA expressions of TrxR and Trx. The mRNA relative expressions of TrxR and Trx in the 26.26 μg／L group were significantly lower than those in the control group ( P＜0.05).2) With the increase of lead concentration, the mRNA relative expressions of HSP60 and HSC70 decreased gradually, and the mR?NA relative expressions of HSP60 and HSC70 in 26.26 μg／L group were significantly lower than those in con?trol group ( P＜0.05). But the mRNA relative expressions of HSP90 increased first and then decreased, and the HSP90 mRNA relative expressions in 13.13 μg／L lead group was significantly higher than that in the other two groups ( P＜0.05). 3) Analysis of intestinal bacteria of all processing groups of M.nipponense at phyla level, three dominant phyla, Proteobacteria, Firmicutes and Tenericutes were observed in the three groups at the phy?lum level in M.nipponense. It was found that the relative abundance of Aeromonas unclassified significantly de?creased after lead stress by the conventional variance analysis approach. Intestinal metagenome was analyzed by LEfSe approach, and it was found that there were differences in intestinal bacteria between the control group and the 26.26 μg／L group. The results showed that the relative abundant of Enterobacteriales, Enterobacteri?aceae, Enterobacter, Citrobacter, Verrucomicrobiae, Verrucomicrobiales, Verrucomicrobiaceae were higher in the control group, and the relative abundant of Deltaproteobacteria, Flavobacteriia, Flavobacteriales, Fla?vobacteriaceae, Flavobacterium, Bacteroidetes, Phreatobacter were higher in the 26.26 μg／L group. Thus it can be concluded that chronic lead exposure decreases the transcription level of Trx and TrxR genes and regu?lates the mRNA relative expressions of HSP60, HSC70 and HSP90 in the hepatopancreas of M.nipponense. Regardless of the concentration of the lead exposure, there are core microorganisms in the intestinal tract of M. nipponense, but high?concentration lead exposure will produce bacteria with significant difference in abun?dance, and the abundance of bacteria to maintain normal metabolism will be reduced, and the bacteria which is related to the degradation of pollutants and the regulation of immune and antioxidant stress will increase.[ Chi?nese Journal of Animal Nutrition, 2019, 31(11) :5378?5390]关键词
热应激蛋白/硫氧还蛋白/肠道菌群/铅/日本沼虾Key words
heat shock protein/thioredoxin/intestinal microbiota/lead/Macrobrachium nipponense分类
农业科技引用本文复制引用
郑纯纯,李小雨,聂欢,周厚杰,丁志丽,田钰滢,杜明川,刘翥..慢性铅胁迫对日本沼虾硫氧还蛋白和热应激蛋白系统基因表达及肠道菌群的影响[J].动物营养学报,2019,31(11):5379-5391,13.基金项目
国家自然科学基金项目(31402308) （31402308）
浙江省自然科学基金项目(LQ14C190004,LY16C190006) （LQ14C190004,LY16C190006）
