Department of Ophthalmology, Kim's Eye Hospital, Myung-Gok Eye Research Institute, Konyang University College of Medicine, Seoul, Korea.
Jpn J Ophthalmol. 2010 Mar;54(2):151-5. doi: 10.1007/s10384-009-0772-6. Epub 2010 Apr 18.
To evaluate the influence of H(2)O(2) on lens epithelial cells (LECs) and to determine the effect of the Janus kinase (JAK) inhibitor AG490 and the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor U0126 on LEC death after H(2)O(2) exposure.
Human lens epithelial (HLE) B-3 cells were cultured. Cells were treated for 45 min with H2O2 and signal transducers and activators transcription (STAT) 3, JAK2, and ERK1/2 phosphorylation were surveyed by Western blot analysis. After pretreatment with either 40 microM AG490 or 25 microM U0126, LECs were exposed to H(2)O(2). LEC death was evaluated by microscopy and flow cytometry.
H(2)O(2) induced phosphorylation of Tyr-705 STAT3, JAK2, and ERK1/2 in LECs. In cells pretreated with both AG490 and U0126, phosphorylation of Tyr-705 STAT3, JAK2, and ERK1/2 was suppressed. Microscopic findings, however, showed that only AG490 noticeably enhanced cell survival, and flow cytometry showed that cell necrosis decreased to 4.05% after pretreatment with 40 microM AG490.
AG490 may prevent H(2)O(2)-induced LEC death by blocking an unknown necrosis pathway. Further investigation is required to characterize that pathway.
评估 H₂O₂对晶状体上皮细胞（LEC）的影响，并确定 Janus 激酶（JAK）抑制剂 AG490 和丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）激酶（MEK）抑制剂 U0126 对 H₂O₂ 暴露后 LEC 死亡的影响。
培养人晶状体上皮（HLE）B-3 细胞。用 H₂O₂处理细胞 45 分钟，通过 Western blot 分析检测 STAT3、JAK2 和 ERK1/2 的磷酸化。用 40 μM AG490 或 25 μM U0126 预处理后，将 LEC 暴露于 H₂O₂中。通过显微镜和流式细胞术评估 LEC 死亡。
H₂O₂诱导 LEC 中 Tyr-705 STAT3、JAK2 和 ERK1/2 的磷酸化。在用 AG490 和 U0126 预处理的细胞中，Tyr-705 STAT3、JAK2 和 ERK1/2 的磷酸化被抑制。然而，显微镜观察结果表明，只有 AG490 明显增强了细胞存活，流式细胞术显示，用 40 μM AG490 预处理后，细胞坏死减少到 4.05%。
AG490 可能通过阻断未知的坏死途径来预防 H₂O₂诱导的 LEC 死亡。需要进一步研究来表征该途径。
