Effects Of Qufeng Huoxue Pill On MiRNA-155,miRNA-146a And MiRNA-182 Expression In Experimental Autoimmune Uveitis Rats’ Retinal Tissue

Object:To explore the efficacy and mechanism of actionof Qufeng Huoxue Pill on experimental autoimmune uveitis(EAU)rats by observing the effects of the Pill on the expression of miRNA-155,miRNA-146 a and miRNA-182 in EAU rats.Method:A total of 65 SPF-class healthy male Lewis rats(130 eyes)were randomly selected according to the table of random digit as the blank group,while others automatically became the model for active immunization and established the EAU rat model,injected to the inner right hind leg of the rats with 35μg emulsifiers,including inter-photoreceptor retinoid binding protein(IRBP)and Freund’s complete adjuvant(FCA).On the 4th day after that procedure,observing the Anterior segment of the rats’ eyes daily under slit lamp microscope,and the inflammation score was performed according to the Caspi score.On the 7th day after immunization,the eyeballs of Lewis rats,which randomly selected from the model rats,were used to make pathological sections and histopathological examinations were performed after HE staining.While completing model,they were randomly divided into model control group,positive control group,lowdose of Qufeng Huoxue pillgroup,high dose of Qufeng Huoxue pill group.After 14 days of continuous intragastric administration,taking the rat retinal tissue and detecting the relative expression quantity of miRNA-155,miRNA-146 a and miRNA-182 by qPCR.Results:1.Validation of the EAU rat model:On the 4th day of modeling,the iris vessels showed Mild-to-moderate hyperemia and dilation,and the ocular inflammation score(1.45±0.74)under a slit lamp microscope;On the 7th day,the ciliary body became hyperemia and the iris vessels were dilated and congested,concurrent with turbidity of anterior chamber,contracted pupil,light reaction subside or disappear.A few number of rats showed hypopyon,occlusio pupillae,and the ocular inflammation score(2.62±0.65)under a slit lamp microscope.Pathological sections of the rat eye showed different degrees of inflammatory cell infiltration in the posterior cornea,iris,ciliary body and retina.The above indicates that the EAU rat model was successfully established on the 7th day after immunization.in the posterior cornea,iris,ciliary body,and retina.The above shows that on the 7th day after immunization,the EAU rat model was successfully established.2.The qPCR test results showed:(1)The relative expression quantity of miRNA-155: Compared with the blank group,the model control group showed significantly high expression,the difference was statistically significant(P<0.01);compared with the model control group,the positive control group and the high-dose group of Qufeng Huoxue Pill were Low expression,the difference was statistically significant(P<0.05);there was no significant difference between the low-dose group of Qufeng Huoxue Pill and the model control group(P>0.01).(2)The relative expression quantity of miRNA-146a: Compared with the blank group,the model control group showed significant high expression,the difference was statistically significant(P<0.01);compared with the model control group,the positive control group showed low expression(P<0.05).The high-dose group of Fenghuoxue Pill was significantly lower(P<0.01),and the difference was statistically significant.There was no significant difference between the low-dose group of Qufeng Huoxue Pill and the model control group(P>0.01).(3)The relative expression quantity of miRNA-182:Compared with the blank group,the model control group showed significantly low expression,the difference was statistically significant(P<0.01);compared with the model control group,the positive control group and the high-dose group of Qufeng Huoxue Pill were Significantly high expression,the difference was statistically significant(P<0.01);the low-dose group of Qufeng Huoxue Pill was not statistically significant compared with the model control group(P>0.01).Conclusion: 1.Established the EAU rat model successfully by using the emulsifiers of IRBP and FCA.2.Qufeng Huoxue Pill can inhibit the expression of miRNA-155 and miRNA-146 a in EAU rat retina,promote the expression of miRNA-182,and regulate the development of EAU by affecting the expression of miRNA-155,miRNA-146 a and miRNA-182 process.3.miRNA-155,miRNA-146 a and miRNA-182 may be used as potential drug targets for the treatment of uveitis.