Effects of Lycium barbarum polysaccharide on genes expression in spleen of immunosuppressed chicks based on transcriptome sequencing

【Objective】 This paper analyzed the effects of Lycium barbarum polysaccharide(LBP)on the gene expression of spleen of immunosuppressed chicks via transcriptome sequencing,and explored the target genes of repair mechanism of LBP on immunosuppression of chicks.【Method】A total of 120 7-day-old Hy-Line brown laying hens were randomly divided into negative control group(NC),cyclophosphamide(CTX)group(CY)and LBP group(CYLbGp). The NC group was intramuscularly injected with normal saline,while the other two groups were intramuscularly injected with 80 mg/(kg·d)CTX for 3 consecutive days for modeling. After modeling,the immunosuppressed chick model was verified by measuring the levels of IL-2,IL-6 and INF-γ in serum. After successful modeling,the CYLbGp group drinking water was added 5 mg/(kg·d)LBP,after the administration of CYLbGp group,spleen was collected to be used for transcriptome sequencing,diferentially expressed genes(DEGs)of P<0.05 and|log₂ Fold Change| ≥ 1 were screened,GO functional annotation analysis and KEGG pathway enrichment analysis were performed to explore the effects of LBP on immunosuppression repair.【Result】 A total of 1049301886 Raw reads were obtained,and 1032097278 Clean reads were recieved after quality control and filtering. There were 701 and 748 DEGs between CY group and NC group and between CYLbGp and CY group respectively. GO functional annotation showed that the significantly returned DEGs after LBP intervention were mainly related to binding,cell process,biological regulation and cellular component functions. The KEGG pathway enrichment results showed that significantly returned DEGs after LBP intervention were mainly related to neuroactive ligand-receptor interaction and retinol metabolism pathway. The key DEGs in the enriched pathway were cholinergic receptor muscarinic 5(CHRM5),cholinergic receptor nicotinic beta 4 subunit(CHRNB4),glutamate metabotropic receptor 1 (GRM1), cholecystokinin (CCK), neuropeptide Y receptor Y1 (NPY1R), cytochrome P450 family 2 subfamily C member 18(CYP2C18),angiotensin II receptor type 1(AGTR1),cytochrome P450 family 3 subfamily A member 5(CYP3A5),serine protease 2(PRSS2)and neurotensin(NTS). The results of RT-qPCR were basically consistent with the trend of transcriptome data,which further indicated that transcriptome data had high reliability.【Conclusion】The repair mechanism of LBP on immunosuppression of chicks may be related to the regulation of neuroactive ligand-receptor interaction and retinol metabolism pathway. Among them,CHRM5,CHRNB4,GRM1,CCK, NPY1R,CYP2C18, AGTR1,CYP3A5,PRSS2 and NTS can be used as target genes in the repair mechanism of LBP on immunosuppression of chicks.