| Objectives To observe the effect of Qingre Mingmu Granule on inflammation and histopathology of anterior segment in experimental autoimmune uveitis(EAU)rats,evaluate the therapeutic effect of Qingre Mingmu Granule on EAU rats,as well as the expression of IL-2,IL-10,IL-17 and IFN-γ protein in the supernatant of spleen and lymph nodes,Objective to explore the possible mechanism of Qingre Mingmu granule in EAU rats.Methods Thirty 30 male Lewis rats were randomly selecteed 5 rats as blank group,after immunization,the remaining 25 rats were randomly divided into model group,Qingre Mingmu granule group(low-dose group,middle-dose group,high-dose group)and Tripterygium wilfordii group,5 rats in each group.The same volume of IRBP R16 peptide fragment was mixed with complete Freund’s adjuvant(CFA)containing 2.5 mg/ml Mycobacterium tuberculosis H37 RA,and then injected subcutaneously into rats for immunization to establish EAU animal model.Meanwhile,the blank group and model group were given distilled water by gavage,the Qingre Mingmu granule group was given Qingre Mingmu granule suspension by gavage,and the Tripterygium wilfordii group was given triptolide suspension by gavage for 7 days.On the 7th day after immunization,the anterior segment of each group was observed under slit lamp every day,and the disease was scored according to the inflammation grade;On the 15 th day after immunization,the unilateral eyeballs of rats in each group were removed for histopathological treatment,Hematoxylin and Eosin(HE)staining sections were examined under microscope,and the scores were made according to the histopathological grading;after unilateral eyeballs were removed,the rats in each group were killed,and the supernatant of spleen and lymph nodes were collected for incubation for 3 days,The expression levels of IL-2,IL-10,IL-17 and IFN-γ proteins in the supernatants were detected by ELISA.Results 1 From the 7th day after immunization,the anterior segment of each group was observed under slit lamp every day.From the 8th day after immunization,the rats in the model group had inflammation reactions such as iris vasodilation,congestion,anterior chamber turbidity,pupil narrowing(membranous closure).The EAU model was successfully established,and the success rate was 100%.2 The inflammation score of the model group increased from the 8th to 12 th day after immunization,which was significantly higher than that of the blank group,and reached the peak on the 13 th to 15 th day,with significant difference compared with the blank group(P < 0.01);on the 9th day after immunization,the inflammation score of the Qingre Mingmu granule group was lower than that of the model group(P < 0.01);on the 13 th day after immunization,the inflammation score of the Qingre Mingmu granule group was higher than that of the model group and the Tripterygium wilfordii group,Compared with the model group,the inflammation score of the Tripterygium wilfordii group was lower(P < 0.01).On the 15 th day after immunization,the inflammation reaction of the Qingre Mingmu granule group was significantly lighter than that of the model group and the Tripterygium wilfordii group,and the inflammation score was significantly lower(P < 0.01).There was no significant difference in the inflammation score of Qingre Mingmu granule group at each time point(P > 0.05).3Histopathological results showed that the degree of retinal inflammatory cell infiltration and damage of retinal tissue structure in Qingre Mingmu granule group was significantly lighter than that in model group and Tripterygium wilfordii group,and the histopathological score was significantly lower than that in model group and Tripterygium wilfordii group(P < 0.01).There was no significant difference in histopathological score of Qingre Mingmu granule group(P > 0.05).4 The results of ELISA showed that: compared with the blank group,the expression of IFN-γ,IL-2 and IL-17 in the supernatant of the model group was significantly increased,while the expression of IL-10 was significantly decreased,the difference was statistically significant(P < 0.01);compared with the model group,the expression of IL-2and IFN-γ in the supernatant of the drug intervention groups was significantly decreased(P< 0.01 or P < 0.05),At the same time,the expression of IL-17 was significantly decreased in Qingre Mingmu granule group(P < 0.01),and the expression of IL-10 was increased in Qingre Mingmu granule group and Tripterygium wilfordii group(P < 0.01 or P < 0.05).Compared with Tripterygium wilfordii group,the expression of IL-2 and IL-17 in Qingre Mingmu granule group decreased(P < 0.01),and the expression of IL-10 increased(P <0.01)Conclusions 1 Qingre Mingmu granule can reduce the retinal tissue damage caused by uveitis,reduce the intraocular inflammatory reaction,and has preventive therapeutic effect on EAU.2 Qingre Mingmu granule can significantly reduce the levels of IRBP specific T cells secreting IL-2,IFN-γ and IL-17,suggesting that Qingre Mingmu granule has antiinflammatory effect and can inhibits the expression of Th1 and Th17 cytokines;meanwhile,the level of IL-10 in the supernatant of IRBP specific T cells intervened by Qingre Mingmu granule is significantly increased,suggesting that Qingre Mingmu granule can promote the secretion of IL-10 and protect the development of EAU.3 Qingre Mingmu granule can improve the severity of autoimmune uveitis in rats,mainly by inhibiting the secretion of proinflammatory effect,which may be related to the regulation of cytokine IL-10,and improve EAU by regulating the relationship between Th1/Th2.Figure 8;Table 6;Reference 158... |
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