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

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

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?

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 lipopdysaccharide (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 contiols 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.