Retinal Transcriptome Analysis In The Treatment Of Endotoxin Induced Uveitis With Tetramethylpyrazine Eyedrops

Purpose: To investigate the role of tetramethylpyrazine(TMP)eye drops in endotoxin induced uveitis(EIU)animal model and the retinal gene expression changes of TMP eyedrops treated and untreated endotoxin induced uveitis mice,and to assess the possible molecular mechanisms by which TMP eyedrops protect against EIU.Methods: EIU were induced by intravitreal injection of 125 ng lipopolysaccharides(LPS).Mice were randomly divided into LPS group and TMP+LPS group.TMP+LPS group was treated with 0.1% TMP eye drops(6 times/24 hours,10 μl each time);LPS group was not treated.24 hours after LPS injection,the inflammatory signs of the anterior segment were examined by a slit lamp and clinical scores were evaluated at the same time by two individuals following the criteria of EIU clinical score.RNA sequencing(RNA-seq)strategy using Illumina Hiseq 2500 platform was applied to identify the retinal transcriptome changes from tetramethylpyrazine(TMP)eyedrops treated and untreated mice.The protein-to-protein interaction(PPI)was analyzed by using the STRINGsoftware.The differentially expressed genes(DEGs)were validated by real time-PCR.Results: The inflammatory responses were much severe and clinical score of LPS group was remarkably higher than the TMP treated group 24 h after LPS administration.RNA-seq identified 407 DEGs,among which 356 were up-regulated and 51 were down-regulated.There were 12up-regulated GO terms enriched and 27 up-regulated pathways.The constructed PPI network consisted of 386 nodes and 2393 edges.Among the 42 networks,the 3 subnetworks are significant.Seven DEGs,including inflammation-related(Ccl12,Ccl2,Gzmb),complement system-related(C1ra)and interferon-related genes(Siglec1,Zbp1 and Ifit1)were validated by using Q-PCR.Conclusion: TMP plays a protective role in endotoxin-induced uveitis,which may be related to some inflammation-related,complement system-related and interferon-related genes,suggesting that inhibiting the expression of these genes may provide a new strategy for the treatment of uveitis.