Effect of Paeoniflorin on Th17/Treg Immune Balance in Experimental Autoimmune Uveitis Mice by Regulating RhoA/ROCK Signaling Pathway
Objective To investigate the effect of paeoniflorin on the immune balance of T-helper 17 lymphocytes(Th17)/regulatory T cells(Treg)in experimental autoimmune uveitis(EAU)mice by regulating the Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing kinase(ROCK)signaling pathway.Methods Ten mice were randomly selected as the control group,and the remaining mice were used to construct an EAU model.The successful modeling mice were randomly divided into model group,paeoniflorin group(10 mg·kg-1 paeoniflorin),LPA group(25 μmol·L-1 LPA,activator of RhoA/ROCK signaling pathway),and paeoniflorin+LPA group(10 mg·kg-1 paeoniflorin+ 25 μmol·L-1 LPA),with 10 mice in each group.The model and control groups were given equal amounts of physiological saline,once a day for 14 days.The clinical minifestation score of the anteior segment of the eye score was applied to assess the severity of inflammation.HE staining was applied to observe the pathological morphology of eyeball tissue.ELISA method was applied to detect the levels of inflammatory factors trarsfornning growth factor beta(TGF-β),interleukin 10(IL-10)and interleukin 17(IL-17)and interleukin-23(IL-23)in serum.Flow cytometry was applied to detect the level of splenic Th17/Treg cells.Western Blot was applied to detect the expression of retinoid-related orphan nuclear receptor γt(RORγt),forkhead box P3(Foxp3),and RhoA/ROCK signaling pathway proteins.Results The mice in the control group had no iris and conjunctival vasodilation and congestion.When compared with the control group,the mice in the model group had iris vasodilation and congestion,and a number of inflammatory cells in the anterior chamber,iris,ciliary body,and retina were found.The clinical minifestation score of the anteior segment of the eye score,the content of IL-17 and IL-23,the level of Th17 cells,Th17/Treg and the protein levels of RORγt,RhoA,and ROCK1 were obviously increased(P＜0.05).The content of IL-10 and TGF-β,the levels of Treg cells and Foxp3 were obviously decreased(P＜0.05).Compared with the model group,the symptoms of iris vascular congestion of mice in paeoniflorin group were obviously alleviated,the clinical minifestation score of the anteior segment of the eye score,the content of IL-17 and IL-23,Th17 cells level,Th17/Treg the protein levels of RORγt,RhoA,and ROCK1 were obviously decreased(P＜0.05).The content of IL-10 and TGF-β,Treg cells level,and Foxp3 level were obviously increased(P＜0.05).However,LPA exacerbated eye damage(P＜0.05),and it reversed the improvement effect of paeoniflorin on EAU mice.Conclusion Paeoniflorin can regulate Th17/Treg immune balance in EAU mice by down-regulating RhoA/ROCK signaling pathway and relieviny uveitis of EAU mice.
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