| Objective: Explore the impact of the QingKaiLing Ophthalmic Gel on Th1and Th2cells in the rats with experimental autoimmune uveitis(EAU) and provide the theoreticalbasis for clinical widely usingMethods:1.To observe the effect of QingKaiLing Ophthalmic Gel on the rats withexperimental autoimmune uveitis(EAU)and compared its effect with the dexamethasoneinjection through the intraperitoneal injection, appraising it effect, for its further applied toclinical uveitis lay the foundation for the prevention and control.EAU was induced in Lewis rats by immunization with IRBP1177–1191in completeFreund’s adjuvant. The rats were randomly divided into three groups.Group A (modelcontrol), group B (QingKaiLing Ophthalmic Gel pointing eye), group C (dexamethasoneinjection through the intraperitoneal injection) and evaluated clinically on days0,7,9,11,13,15,17and evaluated inflammation and pathologically on days9.2. Discuss the impact of the QingKaiLing Ophthalmic Gel on Th1and Th2cells in therats with experimental autoimmune uveitis(EAU)EAU was induced in Lewis rats by immunization with IRBP1177–1191in completeFreund’s adjuvant. The rats were randomly divided into three groups.Group A (modelcontrol), group B (QingKaiLing Ophthalmic Gel pointing eye), group C (dexamethasoneinjection through the intraperitoneal injection). Executed rats on days9and lymphocytesfrom lymph nodes (LNs) and spleen were subjected and cultured that stimulated byIRBP1177–1191to flow cytometry to analyze the frequency of CD4~+cells、CD8~+cells、Interferon γ (IFN-γ) and the ratio of CD4~+/CD8~+. The electrophoresis were used to analyze the expression of IFN-γ and IL-4in peripheral blood on days0,3,5,7,9. Optimizedquantitative real-time RT–PCR was used for consecutive measurement of the relativeexpression of Th1-associated molecules, including interferon γ (IFN-γ) and Th2cells-associated molecules, including interleukin4(IL-4) in Lymph nodes on days0,3,5,7,9.The levels of IFN-γ and IL-4in the supernatants and aqueous humor were determined byenzyme-linked immunosorbent assay.Results:1.To observe the effect of QingKaiLing Ophthalmic Gel on the rats withexperimental autoimmune uveitis(EAU)(1)From the inflammation performance of rat eyes, we found that both B and Cgroups can inhibit the rats occurring uveitis and the difference between the two is small.The inflammation scores of the QingKaiLing Ophthalmic Gel group was（1.2±0.63）gradeand slighter than the model group that the scores was (3.6±0.52) grade. The scores of thedexamethasone group was（1.0±0.67）grade.(2)The result of clinical histopathologic examination(HE stain)The eyes of the control rats were that the outer layer of the retina was extensive andsevere damage with part of the inner retinal damage, many inflammatory cells invasion.This were more serious than B and C group that the performance of the eyes was only afew of inflammatory cells infiltration but no damage. The histopathologic grade of theQingKaiLing Ophthalmic Gel group was （1.5±0.53）grade and slighter than the modelgroup that the scores was (3.3±0.53) grade. The grade of the dexamethasone group was（1.6±0.52）grade.2.The impact of the QingKaiLing Ophthalmic Gel on Th1and Th2cells in the ratswith experimental autoimmune uveitis(1)The result of flow cytometry: The percentages of CD4~+T cells、IFN-γ and theratio of CD4+/CD8+in the lymphocytes from lymph nodes (LNs) and spleen of the group Band C were obviously decreased than group A(P<0.05). The percentages of CD8~+T cellswere obviously increased than group A(P<0.05). The difference between the group B and Ais small(P>0.05).(2)The result of PCR: The expression of IFN-γin the peripheral blood and the lymphnodes of the group B and C were obviously decreased than group A(P<0.05). Theexpression of IL-4were obviously increased than group A(P<0.05). The difference between the group B and A is small(P>0.05).(3) The result of Elisa: The concentration of IFN-γ in the supernatants and aqueoushumor of the group B and C were obviously decreased than group A(P<0.05). Theexpression of IL-4were obviously increased than group A(P<0.05). The differencebetween the group B and A is small(P>0.05).Conclusions:1. QingKaiLing Ophthalmic Gel has obviously treatment to the rats with autoimmuneuveitis and this provides theoretical basis for the widely used in clinical.2. QingKaiLing Ophthalmic Gel can inhibit the activation expression of CD4~+T cellsand promote the expression of CD8~+T cells in the spleen and Lymph nodes.3. QingKaiLing Ophthalmic Gel can influence the activation expression of Th1cellsand Th2cells in the spleen、peripheral blood、aqueous humor and Lymph nodes. |
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