[关键词]
[摘要]
目的:探究炎症介质白细胞介素-10(IL-10)在青光眼引流物植入术后的动态表达,揭示其在术后瘢痕化中的作用。
方法:选取75只新西兰大白兔随机分为3组,球结膜下植入不同的引流物材料,其中PMMA组植入表面涂裹聚一氯对二苯(Parylene C)的聚甲基丙烯酸甲酯(PMMA)块; 硅胶-MMC组植入硅胶块,同时注射丝裂霉素C(MMC); 硅胶组植入硅胶块。收集术后第1、3d,1、2、3、4、8wk兔眼房水,采用ELISA法检测房水中IL-10蛋白表达。收集术后第1、2、3、4、8wk材料块周围结缔组织,采用HE染色检测组织中成纤维细胞增生和炎症细胞浸润情况; 采用免疫组织化学染色及RT-PCR法检测组织中IL-10蛋白及mRNA的表达。
结果:HE染色显示硅胶组中成纤维细胞增生和炎症细胞浸润较PMMA组及硅胶-MMC组显著。ELISA检测显示术后早期三组房水中IL-10表达呈先增高再降低的趋势,术后第3d达高峰; 术后早期(第1d~3wk)硅胶组的表达高于PMMA组和硅胶-MMC组(均P<0.05),术后晚期(第4~8wk)三组表达无明显差异。免疫组织化学及RT-PCR检测显示结缔组织中IL-10蛋白及mRNA的表达在术后第1wk最高,术后第2~3wk逐渐下降,术后第4~8wk再次升高,且硅胶组明显高于其他两组(均P<0.05)。术后晚期(第4~8wk)结缔组织中IL-10蛋白的表达与成纤维细胞增生水平呈正相关性。
结论:青光眼引流物材料植入术后,IL-10表达呈先升高后逐渐降低,4wk后再次升高的动态过程,推测IL-10可能对瘢痕形成起抑制作用。
[Key word]
[Abstract]
AIM:To investigate the dynamic expression characteristics of interleukin-10(IL-10)after implantation of glaucoma drainage material, and to reveal the role of IL-10 on scarring formation.
METHODS:Totally 75 New Zealand white rabbits were randomly divided into three groups, which were implanted with different types of material-Polymethyl methacrylate coated Parylene C(PMMA group), silicone together with injection of Mitomycin C(MMC)(silicon-MMC group)and silicone(silicone group). Aqueous humor were collected at 1, 3d, 1, 2, 3, 4 and 8wk after operation and enzyme-linked immunosorbent assay(ELISA)were utilized to detect the expression of IL-10 in the aqueous humor. The connective tissue surrounding the material were collected at 1, 2, 3, 4 and 8wk postoperatively. Hematoxylin-eosin(HE)staining was applied to evaluate the proliferation of fibroblasts and the infiltration of inflammatory cells. The protein expression and mRNA of IL-10 in the connective tissue were detected by immunohistochemistry and real-time PCR.
RESULTS:Compared with PMMA and silicon-MMC group, silicone group showed significantly increased proliferation of fibroblasts and infiltration of inflammatory cells according to the HE staining result. The result of ELISA showed the expression of IL-10 in the aqueous humor increased significantly at the early stage after surgery, and then decreased gradually,the highest appeared on the third day after operation,and in silicone group there was higher than the other two groups in the early stage postoperatively(1d-3wk)(all P<0.05), and there was no significant difference in the late stages(4-8wk). The protein expression and mRNA of IL-10 in connective tissue were the highest in the first week after operation, decreased gradually at 2-3wk after operation, and increased again at 4-8wk after operation by immunohistochemistry and real-time PCR. And the expression was higher in silicone group than in the other two groups at each time point(all P<0.05). Furthermore, there was a positive correlation between the expression of IL-10 protein and the proliferation of fibroblasts in the late stages(4-8wk).
CONCLUSION: After implantation of glaucoma drainage material, the process of IL-10 increased first, then decreased gradually, and increased again 4wk later, thus IL-10 may be a potential target for inhibiting the scar formation.
[中图分类号]
[基金项目]
安徽省重点研究与开发计划项目(No.202004j07020029); 安徽医科大学临床医学学科建设入库项目(No.2020lcxk029); 安徽医科大学校科研基金项目(No.2021xkj253)