摘要目的 研究大鼠全层巩膜切除眼外滤过术后前房相关性免疫偏离(ACAID)诱导能力的变化及其与术后时间的关系.方法 对照实验研究.50只近交系Wistar大鼠,右眼为实验眼,建立全层巩膜切除眼外滤过术模型后,采用计算机随机数字表法分为5组,每组10只鼠.另外5只SD大鼠,1只Wistar大鼠,提供诱导ACAID所需去上皮角膜片.A组为阴性对照组,在全层巩膜切除眼外滤过术后1周,前房植入Wistar大鼠角膜片;B组为阳性对照组,在全层巩膜切除眼外滤过术后1周,颈部皮下注射SD大鼠脾细胞,前房不植入角膜片;C、D、E组为实验组,分别在全层巩膜切除眼外滤过术后1、4、8周,前房植入SD大鼠角膜片.B组在颈部皮下注射SD大鼠脾细胞悬液,1周后右耳廓皮下注射SD大鼠脾细胞悬液,诱导迟发型超敏反应.其他各组均在前房植入角膜片,2周右耳廓皮下注射SD大鼠脾细胞悬液,诱导迟发型超敏反应.耳廓膨胀指数反映诱导迟发型超敏反应的程度.同时各组取心脏血,检测血清中白细胞介素(IL)4和IL-10浓度,取脾脏组织检测细胞转录因子GATA-3 mRNA表达水平.通过诱导迟发型超敏反应,检测Th2的GATA-3和Th2细胞来源细胞因子IL-4及IL-10,分析机体免疫状态,并进而评价ACAID.此外,各组鼠取心脏血和脾脏组织后,再摘除眼球,进行组织病理学检查,观察前房炎性反应变化.采用SPSS 13.0统计学软件进行数据分析.多组间诱导迟发型超敏反应、IL-4、IL-10及逆转录聚合酶链反应(RT-PCR)检测结果比较采用单因素方差分析,进一步两两比较采用LSD法,各组数据与阳性对照组或阴性对照组比较采用t检验.结果 (1)诱导迟发型超敏反应检测,结果显示全层巩膜切除眼外滤过术后,B、C、D、E组鼠均诱发出迟发型超敏反应.(2)血清IL-4和IL-10浓度检测,结果表明全层巩膜切除眼外滤过术后,3个实验组间血清IL-4及IL-10浓度差异均有统计学意义(F=49.124,6.336;均P<0.01);其中D组鼠血清IL-4及IL-10浓度明显低于C组和E组;血清IL-4及IL-10浓度呈现随术后短时间内上升,而后下降,并随时间推移再次增高的趋势.(3)脾脏组织GATA-3表达检测,结果显示D组和E组鼠脾脏组织中GATA-3表达明显上调.(4)眼球组织病理学检查,可见C组鼠前房内有少量炎性渗出和炎性细胞,D组和E组鼠前房内未见明显炎性渗出和炎性细胞.结论 大鼠全层巩膜切除和眼外滤过术后,将暂时失去诱导ACAID的能力.但随术后前房炎性反应的消退,血清IL-10及IL-4浓度再次增高,脾脏GATA-3表达明显上调,表明机体ACMD的诱导能力有逐渐恢复的趋势.(中华眼科杂志,2009,45:32-37)

abstractsObjective To study glaucoma infiltration surgery induced the anterior chamber associated immune deviation (ACAID) by with or without anterior chamber (AC) implantation of foreign epithelium removed corneal graft in mice. Methods In this study, fifty Wistar mice were randomly divided into five groups (ten in each group) and additional five SD and one wistar mouse were used as the providers of epithelium removed corneal grafts to induce ACAID as follows: Group A: corneal grafts from Wistar mouse were implanted into the AC 1 week after the infiltration surgery. Group B : the spleen cells were injected into the nape of neck 1 week after the glaucoma infiltration surgery. In group C, D, and E, the corneal grafts from SD mice were implanted into the AC at 1 week, 4 week, and 8 week after the glaucoma infiltration surgery, respectively. To induce the delayed type hypersensitivity(DTH), spleen cells were injected into the right ear pinna 1 week after the neck injection in group B, 2 weeks after the AC implantation of corneal grafts in other groups. At the same time points as the induction of DTH, heart blood was collected to detect the concentration of IL-4 and IL-10. The spleens were removed to evaluate the expression of the GATA-3 mRNA by RT-PCR. The eyeballs were enucleated and used to evaluate the histopathoiogical changes. Results (1) The DTH were found in group B, C, D, and E with corneal grafts of SD, but not in group A with that of Wistar. (2)The serum concentrations of IL-4 and IL-10 were statistically different in group C, D, and E (F=49.124,6. 336;P<0.01). The concentrations of IL-4 and IL-10 were siguificantly (P=0.002) lower in group D than those in group C and E (3.759±0.250 vs 5.916±0.500 or 4.566±0.518, and 17.170± 3.943 vs 44.447±17.167 or 35.643±21.233 ug/L, n=10, respectively), which showed a biphase response fluctuation of ILs. (3)The GATA-3 mRNA was obviously up-regulated in group D and E. (4)The exudation and a few inflammatory cells in the AC were observed in group C, but not in group D and E. Conclusions The eye with the glaucoma infiltration surgery results in a temporary incapable of inducing ACAID, which is gradually recovered following the diminish of intraocular inflammation and the increase of levels of IL-4, IL-10 and GATA-3 mRNA. (Chin J Ophtbalmol, 2009,45:32-37)