| Objective: To observe the effect of Qufeng Huoxue Pill on the expression of CD80 and IL-27 of dendritic cells in experimental autoimmune uveitis rats,and to evaluate the therapeutic effect of Qufeng Huoxue Pill on EAU rats and explore its possible mechanism.Methods: According to the method of random number table,12 of 48(96eyes)SPF male Lewis rats were randomly selected as healthy control group(group A).The remaining 36 rats were injected subcutaneously with Freund's complete adjuvant(Completefreund'sadjuvant,CFA)containing retinal photoreceptor interreceptor vitamin A binding protein(inter-photoreceptorretinoidbindingprotein,IRBP)1177 color 1191 into the inner hind leg to establish the EAU rat model.After the establishment of the model,the expression of anterior ganglitis in rats was observed by hand-held slit lamp microscope every day,and the inflammation was scored according to the Caspi score standard.After 7 days of active immunization,3 immunized Lewis rats were randomly selected,and 1 rat was randomly selected from the healthy control group.The eyeballs were removed to make retinal pathological sections,and HE staining was performed for histopathological examination.After successful modeling,EAU rats were randomly divided into model control group(group B),low dose Qufeng Huoxue pill group(group C)and high dose Qufeng Huoxue pill group(group D),with 11 rats in each group.The rats were given intragastric administration continuously for 14 days.After anaesthesia,the spleen of rats was isolated.The relative expression of CD80 of dendritic cells in spleen was detected by flow cytometry,and the expression of IL-27 in serum was detected by enzyme-linked immunosorbent assay.Results:1.Verification of EAU rat model: After immunization,the anterior segment of both eyes of rats in the model group was observed underhand-held slit lamp microscope every day,and on the third day,slight inflammatory reaction was observed under slit lamp microscope,showing slight dilation of iris surface vessels,corneal limbal vascular congestion,partial pupil contraction,and ocular inflammation score(0.69 ±0.25).On the 5th day,corneal edema,pupil narrowing,iris hyperemia and edema,unclear texture,different degrees of anterior chamber opacity,posterior corneal wall deposition,aqueous humor glow,decreased fundus red light reflex and ocular inflammation score(1.94 ±0.23)were seen on the 5th day.On the 7th day,there was obvious inflammation in the anterior segment,obvious dilatation and hyperemia of iris vessels,severe opacity of anterior chamber,turbid aqueous humor,closure of pupil membrane and gray-white flocs of pupil margin,showing significant uveitis reaction,and the score of ocular inflammation was(2.97±0.29).Seven days after active immunization,the pathological sections of the eyeball of rats showed different degrees of inflammatory cell infiltration in the posterior cornea,iris,ciliary body and retina.The above results show that the EAU model of Lewis rats was established successfully.2.The results of flow cytometry showed that: The expression of CD80 was detected by flow cytometry: compared with the healthy control group,the expression of CD80 in the model control group was higher,the expression of CD80 in the low-fold group was lower than that in the model control group,and the expression of CD80 in the high-fold group was significantly lower than that in the low-fold group(P<0.01).Compared with the low-fold group of Qufeng Huoxue pill,the expression of CD80 in the high-fold group was significantly lower than that in the high-fold group(P<0.01).3.The level of IL-27 in serum was detected by ELISA method:compared with the healthy control group,the expression of IL-27 in the model control group was high,compared with the model control group,the expression of IL-27 in the low-fold group was lower than that in the high-fold group,and the expression of IL-27 in the high-fold group wassignificantly lower than that in the high-fold group(P<0.01),but there was no significant difference between the healthy control group and the high-fold group.There was no statistical difference between Qufeng Huoxue Pill low power group and Qufeng Huoxue Pill high power group.Conclusion: 1.Through IRBP antigen-Freund's complete adjuvant method,active immunization of Lewis rats can successfully establish EAU rat model.2.On the one hand,Qufeng Huoxue Pill can down-regulate the expression of CD80 in splenic dendritic cells of EAU rats,indicating that DCs may be the target of Qufeng Huoxue Pill on EAU rats,so as to improve the pathological process of EAU through dendritic cells.On the other hand,it also down-regulates the expression of IL-27 in the serum of EAU rats,indicating that Qufeng Huoxue Pill may play an anti-inflammatory effect by inhibiting the initial antigen presenting cells of autoimmune uveitis,thereby inhibiting the differentiation of T cells to Th1 and reducing the expression of IL-27. |
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