The Difference Of Gene Expression Profiles In Uterine Body Of Rabbit After LPS-induction Is Revealed By RNA-sequencing

Genital tract infection female animal is common disease which impair on animal health and decrease reproductive productivity of female animals.Gram negative bacterium is the most pathogenic bacteria in the genital tract infection,and these infections seriously threaten reproductive function of female animals.LPS is a gram negative pathogen associated molecular pattern(PAMP),and its simulated infection causes the body to have the same inflammatory response as gram negative bacteria.LPS administration instead of Gram negative bacteria infection and the transcriptome changes in the uterine body after LPS-induction,still don't be reported.In this study,the transcriptome was sequenced to obtain the changes in m RNA expression profiles in the rabbit uterine body before and after LPS-induction,and to reveal the mechanism of LPS-induced inflammation in the rabbit uterus,which laid the foundation for the treatment of the uterus inflammation caused by gram-negative bacterium.Six female New Zealand white rabbits(b.w.:2.20 � 0.56 kg,month old: 6month old)in the estrous were randomly dividle into two groups,and LPS group(n=3)was intravaginally perfused with 4mg/Kg b.w.of LPS,and the control group(n=3) was perfused with the corresponding volume dose of saline.The tissue of uterine body were collected by 3h of post-perfusion,and was applied to transcriptome sequencing and q RT-PCR analysis.The results showed that: 1)the comparisons of Clean Data and reference genome was carried out,and the mapping efficiency ranged from 68.68% to 74.81%.2)16980 transcripts were identified in the 6 transcriptional libraries,among which 11173 transcripts were expressed in LPS group and control group.There were 294 transcripts specifically expressed in LPS group,and 287 in control group.3)283 differentially expressed genes were screened,among them,160 gene were up-regulated and 123 genes were down-regulated.209 differentially expressed genes were annotated to 62 GO terms and 88 differentially expressed genes were annotated to 50 KEGG pathway.4)the results of q RT-PCR showed that there was a high correlation between q RT-PCR and RNA-seq(r=0.84).5)the expression of inflammation related genes,ADCY8,ADORA2 B,HTR2B,PTGIS,AGT,APOD,PLA2G4 D,SERPINE1 and ENSOCUG00000029731 in the rabbit uterine body were up-regulated,and AIF1 L,CHI3L1,CXCL10,RLA-DRB1,AGTR2 were down-regulated.6)MAPK signaling pathway is important pathway wchich involved immune response in rabbit uterine body,and T lymphocyte and macrophage induced pathways also involved in the process of immune stress.Conclusion: LPS-stimulation induce immunoresponse in the uterine body of rabbit,and cytokine secretory increased.The expression levels of cytokines IL-10,IL-8,INF-gamma and TNF-alpha were inhibited after LPS-stimulation in the uterine body of rabbit.MAPK signaling pathway may be the most important immune response pathway,and T lymphocytes and macrophages also participate in the immunresponse process.