The Identification Of Heat Stress-related Lnc RNA In Cows And The Functional Study Of LncRNA TCONS₀0014079

Heat stress affects the performance and mammary health of cows,which brings great challenge to the dairy industry.Long non-coding RNA(lnc RNA)has been proved to be involved in a variety of life activities.To explore the influence of heat stress on bovine mammary gland tissue lnc RNA expression,by using high-throughput sequencing,we obtained the heat stress cows and the non heat stress stress of bovine mammary gland tissue lnc RNA expression spectrum.In order to explore the role of lnc RNA in the heat stress of dairy cows,we analysed differentially expressed lnc RNAs and verified the function of lnc RNA.The main research contents and results are as follows:1.Effect of heat stress on lnc RNA in mammary gland of dairy cows: The mammary gland tissues of 6 heat stress cows and non-heat stress cows were collected respectively,and the expression profile of mammary gland lnc RNA was identified by high-throughput sequencing technology.The results showed that 1003 lnc RNAs were identified.A total of 34 lnc RNAs were differentially expressed,among which 28 lnc RNAs were down-regulated in the heat stress group and 6 lnc RNAs were up-regulated in the heat stress group.Go enrichment and KEGG pathway analysis of target genes of differentially expressed lnc RNAs found that the major metabolic pathways involved in Nucleotide excision repair,NF kappa B signaling pathway,Cell adhesion and Immune system.2.Functional verification of lnc RNA: Eight differentially expressed lnc RNAs were randomly selected for fluorescent quantitative PCR verification.The relative expression data was consistent with the sequencing results,which proved that the high-throughput sequencing results had high reliability.Differential expression of lncrna tcons?00014079 was selected as a candidate lnc RNA for functional verification because of its large difference multiple and high fpkm value.The prediction of target gene function showed that lnc RNA TCONS?00014079 may be involved in the regulation of inflammatory response,so in this study,the establishment of bovine mastitis cell model and si RNA was used to explore the effect of lnc RNA TCONS?00014079 on bovine mastitis.The results showed that lnc RNA TCONS?00014079 was down-regulated in the bovine mastitis cell model,and after si RNA transfection,it was found that the expression of TNF-?,IL-6 and IL-8,which were originally up-regulated in bovine mastitis cell model,decreased significantly,suggesting lnc RNA TCONS?00014079 may be involved in the inflammatory response of bovine mammary gland by regulating the expression of inflammatory factor m RNA.In conclusion,we found that there were different expression of lnc RNA in the mammary gland of heat stressed and non heat stressed dairy cows,and the selected lnc RNA TCONS?00014079 is involved in the inflammation of mammary gland by influencing the expression of TNF-?,IL-6 and IL-8.