Study On Hepatopancreas MicroRNA Expression Profiles Of Sinopotamon Henanense After Cadmium Exposure

In recent years,food safety,as an important issue of people's livelihood,has attracted wide public attention.With the rapid economic growth and the continuous development of modern industry,the pollution of heavy metal cadmium in aquatic organisms has become more serious,which has a huge impact on the food safety of aquatic products and has caused various health hazards to humans.Sinopotamon henanense is widely distributed in China,and it is very sensitive to heavy metals.It is an ideal indicator for monitoring aquatic organisms' heavy metal cadmium pollution.By studying the toxic effects of cadmium on Sinopotamon henanense and its molecular mechanism,it is helpful to explore the harmful effects of cadmium pollution on the production and reproduction of aquatic organisms and food safety of aquatic products.Hepatopancreas is the main organ for cadmium accumulation in Sinopotamon henanense,and the main target organ of cadmium toxicity.The research team previously revealed the toxic effects of cadmium on the hepatopancreas of Sinopotamon henanense through biochemistry,cell biology and other methods.However,its molecular mechanism is poorly understood,especially the post-transcriptional regulation mechanism of cadmium pollution on the Sinopotamon henanense.In addition,in order to better simulate the growth environment of aquatic organisms,this study adopted a subchronic cadmium exposure method and selected a low concentration(0.5mg/L)as the cadmium treatment concentration.Sinopotamon henanense were randomly divided into two groups: the control group and the experimental group(0.5mg/ L).After 30 days of treatment,the hepatopancreas were taken for the following experiments.(1)Determination of hepatopancreas micro RNA(mi RNA)sequence of Sinopotamon henanense It was subjected to RNA extraction,c DNA library construction,original sequence acquisition,sequence data screening,and mi RNA analysis.The results showed that 13543113 and 11437298 original sequences were obtained from the two libraries of the experimental group and the control group,of which 10387853 and 9687351 were high-quality sequences.By comparison with the mi Rbase database,a total of 656 mi RNAs were obtained in the libraries of the control and experimental groups,including 465 known mi RNAs and 191 newly discovered mi RNAs.Among the known mi RNAs,Sinopotamon henanense shared a total of 24 identical sequences with Portunus trituberculatus,Eriocheir sinensis and Scylla paramamosain.(2)Identification of differentially expressed mi RNA in hepatopancreas control group and experimental group of Sinopotamon henanense and verification by quantitative real-time PCR In this study,mi RNAs that were differentially expressed in the hepatopancreas of the experimental group and the control group of Sinopotamon henanense were screened.By q RT-PCR,in order to further confirm the correctness of sequencing results,the results obtained were verified.The results showed that there were 51 differentially expressed mi RNA molecules between the experimental group and the control group.Eight mi RNAs were randomly selected for q RT-PCR verification,and the quantitative analysis results of seven mi RNAs were completely consistent with the results of deep sequencing analysis.(3)Prediction of differentially expressed mi RNA molecules bioinformatics functions in hepatopancreas of cadmium-exposed group of Sinopotamon henanense Bioinformatics function prediction was performed on 51 differentially expressed mi RNAs screened earlier.The results showed that the 51 differentially expressed mi RNAs in the hepatopancreas cadmium exposure group of Sinopotamon henanense had a total of 5258 target genes.Geno Ontology(GO)analysis showed that these target genes were related to a total of 386 functional classes,of which 17 had significant differences.These 17 functions were mainly related to the activity of oxidoreductase.The results of metabolic pathway analysis showed that they were related to 185 metabolic signaling pathways,of which 18 metabolic signaling pathways were significantly different,such as m RNA stability,N-glycan biosynthesis,and cell endocytosis.The following conclusions were reached through experimental research:(1)A total of 656 mi RNAs were detected in the control group and the cadmium-treated group by mi RNA sequencing,including 465 known mi RNAs and 191 newly discovered mi RNAs.(2)Between the control group and the cadmium-treated group,51 mi RNAs were differentially expressed.The q RT-PCR verification results showed that the accuracy and reliability of sequencing results were high.(3)The results of GO gene function analysis showed that most of the targeted genes were related to the regulation of oxidoreductase activity.KEGG metabolic pathway analysis results showed that most of the targeted genes were related to protein biosynthesis,degradation and modification.The above studies are helpful to explore the molecular mechanism of cadmium on the hepatopancreas toxicity of Sinopotamon henanense,and to provide basic data for exploring the toxic effects of cadmium on aquatic organisms and the toxicity hazards of aquatic food safety.