目的：探究炎症因子白介素-8(IL-8)影响后发性白内障(PCO)细胞迁移的过程中对晶状体上皮细胞(LEC)分泌的单核细胞趋化蛋白1(MCP-1)的调控作用。

方法：构建大鼠晶状体囊袋模型，采用10%胎牛血清培养撕囊后的晶状体囊袋使LEC迁移至后囊30%-50%面积后，撤去血清，将囊袋分为对照组和15 ng/mL IL-8组，并在不同时间点进行拍照观察囊袋LEC迁移。ELISA法和RT-qPCR检测不同组MCP-1的分泌和信使RNA(mRNA)相对表达水平。免疫荧光检测不同组MCP-1表达水平。将SRA01/04细胞分为对照组、15 ng/mL IL-8组和15 ng/mL IL-8+200 μmol/L Bindarit(BND)组，用Transwell法检测不同组迁移细胞数量。将SRA01/04细胞分为阴性对照(NC)组、NC+15 ng/mL IL-8组及15 ng/mL IL-8+p65 siRNA组，ELISA法和RT-qPCR检测不同组MCP-1的分泌和mRNA相对表达水平。

结果：体外培养的大鼠晶状体囊袋LEC迁移水平显示，在48、72、96 h时15 ng/mL IL-8组囊袋内细胞迁移明显增加(均P<0.05)。ELISA的结果显示，15 ng/mL IL-8组囊袋和SRA01/04细胞与对照组相比较，在12、24 h分泌的MCP-1的水平升高(均P<0.05)； RT-qPCR的结果同样显示15 ng/mL IL-8组SRA01/04细胞在12、24 h分泌的MCP-1 mRNA的相对表达水平增高(均P<0.05)。免疫荧光的结果显示，与对照组相比，24 h时15 ng/mL IL-8组的囊袋上皮细胞MCP-1表达水平升高(P=0.007)。Transwell的检测结果显示，与对照组相比，15 ng/mL IL-8组迁移数量增加(P=0.001)； 与15 ng/mL IL-8组相比，15 ng/mL IL-8+200 μmol/L BND组迁移数量减少(P=0.003)。ELISA和RT-qPCR的结果显示，与NC组相比，NC+15 ng/mL IL-8组在12、24 h时的MCP-1分泌和mRNA的相对表达增加(均P<0.01)； 与NC+15 ng/mL IL-8组比较，15 ng/mL IL-8+p65 siRNA组在12、24 h时的MCP-1分泌和mRNA的相对表达减少(均P<0.01)。

结论：IL-8可促进囊袋内残留上皮细胞的迁移，调控晶状体上皮细胞MCP-1的分泌和表达水平，推测其作用机制与NF-κB/p65信号通路有关。

METHODS: A rat lens capsule model was established and cultured in medium supplemented with 10% fetal bovine serum. Upon migration of LEC to 30%-50% of the posterior capsule, serum was removed. The capsule was subsequently divided into two groups: a control group and an IL-8(15 ng/mL)treatment group. LEC migration was captured at multiple time points. The secretion and mRNA expression of MCP-1 were quantified using ELISA and RT-qPCR, respectively. Immunofluorescence was used to assess MCP-1 expression in the different experimental groups. SRA01/04 cells were divided into three groups: control, IL-8(15 ng/mL), and IL-8(15 ng/mL)+200 μmol/L Bindarit(BND)groups, with migration measured by the Transwell assay. Additionally, SRA01/04 cells were divided into negative control(NC), NC+15 ng/mL IL-8, and 15 ng/mL IL-8+p65 siRNA groups, and MCP-1 secretion and mRNA expression were further analyzed by ELISA and RT-qPCR.

RESULTS:LEC migration in the rat lens capsule cultured in vitro showed that the cells migration of the 15 ng/mL IL-8 group significantly increased at 48, 72 and 96 h(all P<0.05). ELISA results revealed that MCP-1 levels in SRA01/04 cells from the 15 ng/mL IL-8-treated group were markedly higher than those in the control group at both 12 and 24 h(all P<0.05). RT-qPCR analysis also demonstrated a significant increase in MCP-1 mRNA expression in the 15 ng/mL IL-8 group at both time points(all P<0.05). Immunofluorescence staining indicated greater MCP-1 expression in capsular epithelial cells of the 15 ng/mL IL-8 group at 24 h(P=0.007). Transwell assays further confirmed increased cell migration in the 15 ng/mL IL-8 group compared to the control group(P=0.001), while the migration reduced in the 15 ng/mL IL-8+200 μmol/L BND group compared to the 15 ng/mL IL-8 group(P=0.003). Moreover, ELISA and RT-qPCR results demonstrated a significant increase in MCP-1 secretion and mRNA expression in the NC+15 ng/mL IL-8 group at both 12 and 24 h compared to the NC group(all P<0.01). In contrast, MCP-1 secretion and mRNA expression were reduced in the 15 ng/mL IL-8+p65 siRNA group compared to the NC+15 ng/mL IL-8 group at both time points(all P<0.01).

CONCLUSION: IL-8 promotes the migration of residual epithelial cells and regulates the secretion and expression of MCP-1 in LEC. The mechanism underlying IL-8's effects appears to be mediated through the activation of the NF-κB/p65 signaling pathway.