【摘要】 目的:探讨缺氧条件下Wnt/β-catenin通路在晶状体上皮细胞上皮-间充质转化（EMT）中的作用，分析Dickkopf-1（DKK-1）的表达对晶状体上皮细胞EMT的影响。方法:将体外培养人晶状体上皮细胞（HLEB3细胞）分为正常氧培养组（加入含10%FBS的DMEM培养液）和缺氧培养组（加入含100μmol/L CoCl₂溶液的培养液进行缺氧处理6、12、24、48h）。利用免疫荧光染色法检测Wnt3a和DKK-1蛋白的表达及β-catenin蛋白的表达和定位；利用qRT-PCR法检测DKK-1 mRNA的表达。结果:免疫荧光染色结果显示，随着缺氧时间的延长，HLEB3细胞中Wnt3a和DKK-1蛋白表达水平明显上调，β-catenin蛋白在细胞核内积聚逐渐增多。qRT-PCR检测结果显示，与正常氧培养组相比较，缺氧培养6h组细胞中DKK-1 mRNA无显著差异（P>0.05）,而缺氧培养12、24、48h组细胞中DKK-1 mRNA表达明显增加（P<0.001）。结论:缺氧可以激活晶状体上皮细胞Wnt/β-catenin通路，并诱导Dickkopf-1的表达，参与调控Wnt/β-catenin通路，影响EMT进程。更多还原

【Abstract】 ?AIM: To scrutinize the role of the Wnt/β-catenin signaling pathway in the epithelial-mesenchymal transition(EMT) of lens epithelial cells under hypoxic conditions, and to further analyze the effect of Dickkopf-1(DKK-1) expression on EMT of lens epithelial cells.?METHODS: Human lens epithelial cells(HLEB3 cells) were propagated in vitro and then separated into two groups: one exposed to standard oxygen levels, added DMEM culture solution containing 10% FBS(normoxic group) and another subjected to low oxygen levels(hypoxic group). The hypoxic condition was emulated by applying a concentration of 100 μmol/L cobalt chloride(CoCl2) for 6, 12, 24, and 48h. The utilization of immunofluorescence staining enabled the detection of Wnt3a and DKK-1 expressions, along with the expression and localization of β-catenin protein in these groups. The expression of DKK-1 mRNA was discerned by quantitative real-time polymerase chain reaction(qRT-PCR).?RESULTS: Immunofluorescence assays indicated an escalating trend in the Wnt3a and DKK-1 protein expression, which corresponded with the increasing duration of hypoxia. Likewise, an intensified nuclear accumulation of β-catenin protein was observed to be directly proportional to the length of hypoxia treatment. The qRT-PCR demonstrated that the difference in DKK-1 mRNA expression between the normoxic group and the group exposed to hypoxia for 6h was not statistically significant(P>0.05), whereas the DKK-1 mRNA expression of the 12, 24, and 48h hypoxia groups were significantly increased(P<0.001).?CONCLUSION: Hypoxia can activate Wnt/β-catenin pathway in lens epithelial cells and induce the expression of DKK-1, thus regulating the Wnt/β-catenin pathway and affecting the EMT process.更多还原