龙胆泻肝汤对葡萄膜炎大鼠关键炎症细胞因子表达的影响

Immunomodulatory effects of Longdan Xiegan Tang on related inflammatory cytokines in rats with experimental autoimmune uveitis

目的探讨龙胆泻肝汤对实验性自身免疫性葡萄膜炎(experimental autoimmune uveitis,EAU)大鼠关键炎症细胞因子表达的影响。方法 Lewis大鼠随机分为对照组(6只)、EAU组(18只)和LXT组(18只)。光感受器间维生素A结合蛋白(interphotoreceptor retinoid-binding protein,IRBP)免疫EAU组和LXT组大鼠,对照组大鼠注射等量不含IRBP的乳化液。使用实时荧光定量PCR及ELISA方法检测血液、淋巴结和脾脏中IFN-γ、IL-17、TNF-α和IL-10细胞因子在EAU大鼠体内治疗前后的变化情况。结果 LXT组大鼠全血中IFN-γmRNA表达在第8天虽然高于对照组(P=0.000),但已显著低于EAU组(P=0.000);LXT组IL-17 mRNA在第12天达峰值,显著低于EAU组(P=0.000),且均显著高于对照组(P=0.000);TNF-αmRNA仅在第12天显著高于对照组(P=0.000),但低于EAU组(P=0.000);IL-10 mRNA表达在第16天高于EAU组(P=0.042)。LXT组大鼠淋巴结和脾脏中IFN-γ、IL-17和TNF-αmRNA表达在第12天显著低于EAU组(均为P=0.000);LXT组淋巴结中IL-10 mRNA表达在第8天开始显著高于对照组(P=0.001),且在整个炎症期均维持在较高水平;LXT组脾脏IL-10 mRNA表达在第12天达峰值,且高于对照组和EAU组(均为P=0.000)。LXT组大鼠血清中IFN-γ、IL-17和TNF-α蛋白表达水平在第12天和第16天均低于EAU组(均为P<0.05);EAU组和LXT组血清中IL-10蛋白表达在第12天均高于对照组(均为P<0.05),第16天达峰值,但LXT组IL-10蛋白含量更高。结论龙胆泻肝汤通过抑制IFN-γ、IL-17和TNF-α相关炎症因子的表达发挥其抗炎作用,促进IL-10的表达,加速自身免疫炎症的恢复。龙胆泻肝汤可通过多种途径在EAU的治疗中发挥免疫调节作用。

Objective To study the immunomodulatory effects of Longdan Xiegan Tang on related inflammatory cytokines in rats with experimental autoimmune uveitis （EAU）. Methods Lewis rats were randomly divided into control group （5 rats）, EAU group （ 18 rats） and LXT group （ 18 rats）. Rats in EAU and LXT groups were im- munized with interphotoreceptor retinoid-binding protein （IRBP） emulsion. The expressions of IFN-γ,IL-17,IL-10 and TNF-α were investigated by quantitative real-time PCR and ELISA, respectively. Results In blood of LXT group, the mRNA level of IFN-γon day 8 was higher than that in control group （P = 0. 000）, but obviously lower than that in EAU group （P =0. 000 ） ; The mRNA level of IL-17 was peaked on day 12,but was lower than EAU group （P = 0. 000） ; The mRNA level of TNF-α on day 12 was higher than that in control group （ P = 0. 000 ）, but obviously lower than that in EAU group （P =0. 000） ; The mRNA level of IL-10 on day 15 was higher than that in EAU group （P=0. 042）. In lymph node and spleen of LXT group,the mRNA levels of IFN-γ,IL-17 and TNF-α on day 12 were lower than those in EAU group （ all P =0.000 ） ; The mRNA level of IL-10 was peaked on day 12,but was lower than EAU group （P =0. 000）. In serum of LXT group,the mRNA levels of IFN-γ,IL-17 and TNF-α on day 12 and day 15 were lower than those in EAU group;The level of IL-10 in EAU and LXT group on day 12 were higher than that in control group,peaked on day 15,but the LXT group was the highest. Conclusion Longdan Xiegan Tang can reduce the expressions of proinflammatory cytokines including IFN-γ,IL-17 and TNF-α. Meanwhile,it can also promote the production of IL-10 and further accelerate the recovery of EAU,indicating that Longdan Xiegan Tang can play a significant role in treating uveitis.