伤寒杆菌内毒素对GATA-3表达及前房相关免疫偏离形成的影响

目的　观察眼部炎症时前房注射抗原后GATA 3表达的变化以及对前房相关免疫偏离(ACAID)形成的影响。方法　将 5 μl光感受器间维生素A类结合蛋白 (IRBP) ( 10g/L)注射至SD大鼠前房诱导ACAID的动物模型 ,于前房注射IRBP前 4d( - 4d组 )、2 4h( - 2 4h组 )、0h( 0h组 )以及前房注射IRBP后 3d( 3d组 )、7d( 7d组 )从足底注射 2 0 0 μg伤寒杆菌内毒素 (LPS)。各组于前房注射IRBP后第 8天 ,采用酶联免疫吸附分析 (ELISA)法检测血清白细胞介素 4 (IL 4 )水平 ,以监测ACAID的形成 ,并取脾组织行逆转录 聚合酶链反应 (RT PCR)和Western免疫印迹 ,检测GATA 3基因及蛋白表达的变化 ,进行半定量分析。每组 6只 ,每个样品实验重复 3次。结果　 - 2 4h组和 0h组LPS注射后 2 4h眼部炎症到达高峰 ,前房注射IRBP 8d后血清中测不出IL 4 ,GATA 3基因和蛋白表达无明显增加 ;- 4d组LPS注射 4d后眼部炎症消退 ,此时前房注射IRBP ,8d后血清中IL 4水平显著升高( 10 4pg/ml± 3 4pg/ml,P <0 0 1) ,GATA 3基因和蛋白表达增加 ;3d组前房注射IRBP 8d后血清中未测出IL 4 ,GATA 3基因和蛋白表达无增加 ;7d组前房注射IRBP 8d后血清中IL 4升高 ( 10 0pg/ml± 2 5pg/ml,P <0 0 1) ,GATA 3基因和蛋白表达增加。结论　LPS诱导眼部炎

Effect of endotoxin-induced uveitis on GATA-3 expression and ACAID development

OBJECTIVE: To investigate the changes of the GATA-3 expression and development of anterior chamber associated immune deviation (ACAID) after anterior chamber (AC) injection of interphotoreceptor retinoid binding protein (IRBP) under ocular inflammation. METHODS: ACAID was induced by injection of IRBP into the AC of 30 Spar-Dawley (SD) rats. Then the animals were divided into -4 days group, -24 hours group, 0 hour group, 3 days group, and 7 days group according to the lipopolysaccharide (LPS) injection 4 days and 24 hours before, or 0 hour, 3 days and 7 days after IRBP inoculation respectively. 6 rats were used as controls 8 days after IRBP injection, Serum interleukin-4 (IL-4) was evaluated to count the development of ACAID; Western blot and RT-PCR were used to determine the protein and mRNA levels of GATA-3 expression. RESULTS: In -24 hours group and 0 hour group, the ocular inflammation reached a maximum 24 hours after LPS injection; on 8 days after IRBP inoculation, serum IL-4 couldn't be detected and GATA-3 expression were not changed both on mRNA and protein levels compared with control group. In -4 days group, the ocular inflammation was subsided gradually 24 hours and disappeared 96 hours after LPS injection; serum IL-4 and GATA-3 expression were significantly elevated 8 days after IRBP injection. In 7 days group, the serum IL-4 and GATA-3 expression in spleen increased 8 days after IRBP inoculation. CONCLUSION: In ocular inflammation, the up-regulation of GATA-3 expression is inhibited and ACAID development is blocked after antigen was injected into anterior chamber. Once GATA-3 is up-regulated, LPS injection cannot affect ACAID development.