摘　要：为探究隐丹参酮对过氧化氢（H2O2）诱导的人晶状体上皮细胞（HLE-B3）对细胞氧化应激、增殖、凋亡以及c-Jun氨基末端蛋白激酶（JNK）通路的调控作用，体外培养HLE-B3细胞系，用100 μmol/L H2O2干预HLE-B3 24 h，再用不同浓度的隐丹参酮（2.5、5.0、10.0 μmol/L）分别处理后，根据HLE-B3细胞活力结果筛选最适隐丹参酮浓度。细胞再分为对照组（不做干预处理）、H2O2组（100 μmol/L H2O2处理）、隐丹参酮组（100 μmol/L H2O2+10.0 μmol/L隐丹参酮处理）、SP 600125组（100 μmol/L H2O2+20 μmol/L JNK信号通路抑制剂SP 600125处理）、抑制剂组（100 μmol/L H2O2+10.0 μmol/L隐丹参酮+20 μmol/L SP 600125处理）和激活剂组（100 μmol/L H2O2+10.0 μmol/L隐丹参酮+2 mg/L JNK信号通路激活剂茴香霉素处理），药物干预处理均为24 h。细胞计数试剂盒-8（CCK-8）测定细胞活力；试剂盒法检测超氧化物歧化酶（SOD）和丙二醛（MDA）的含量；5-乙炔基-2′脱氧尿嘧啶核苷（EdU）法检测细胞的增殖能力；Hoechst 33258染色试剂盒测定细胞凋亡能力；蛋白免疫印迹（Western blot）法测定细胞周期蛋白D1（CyclinD1）、半胱氨酸蛋白酶-3（Caspase-3）及JNK通路相关蛋白表达水平。根据HLE-B3细胞活力结果选择10.0 μmol/L隐丹参酮用于后续试验。H2O2组SOD水平、增殖率和CyclinD1蛋白水平低于对照组（P<0.05），MDA水平、凋亡率、Caspase-3和磷酸化（p）-JNK蛋白水平高于对照组（P<0.05）；隐丹参酮组和SP 600125组显著抑制了H2O2对HLE-B3细胞的上述作用（P<0.05）；与隐丹参酮组比较，抑制剂组增强了隐丹参酮在H2O2诱导的HLE-B3细胞中的作用（P<0.05），激活剂组则削弱了隐丹参酮在H2O2诱导的HLE-B3细胞中的作用（P<0.05）。隐丹参酮通过抑制JNK信号通路抑制H2O2诱导的HLE-B3氧化应激和凋亡，促进其增殖。
关键词：白内障；人晶状体上皮细胞；隐丹参酮；过氧化氢；c-Jun氨基末端激酶信号通路；氧化应激
中图分类号：R774                          文献标志码：A                 DOI：10.3969/j.issn.1007-7146.2025.01.011

Abstract: To explore the regulatory effects of cryptotanshinone on oxidative stress, proliferation, apoptosis and c-Jun amino-terminal protein kinase (JNK) pathway in human lens epithelial cells (HLE-B3) induced by hydrogen peroxide (H2O2). HLE-B3 cell line was cultured in vitro, and HLE-B3 was treated with 100 μmol/L H2O2 for 24 h, and then treated with different concentrations of cryptotanshinone (2.5, 5.0, 10.0 μmol/L) respectively. The optimal concentration of cryptotanshinone was selected according to the results of HLE-B3 cell viability. Then the cells were divided into control group (no intervention), H2O2 group (100 μmol/L H2O2 treatment), cryptotanshinone group (100 μmol/L H2O2+10.0 μmol/L cryptotanshinone treatment), and SP 600125 group (100 μmol/L H2O2+20 μmol/L JNK signaling pathway inhibitor SP 600125), inhibitor group (100 μmol/L H2O2+10.0 μmol/Lcryptotanshinone +20 μmol/L SP 600125) and activator group (100 μmol/L H2O2+10.0 μmol/L cryptotanshinone+2 mg/L anisomycin, JNK signaling pathway activator), and drug interventions were performed for 24 hours. The contents of superoxide dismutase (SOD) and malonaldehyde (MDA) were detected by kit method. 5-ethynyl-2′-deoxyuridine (EdU) assay was used to detect cell proliferation. Hoechst 33258 staining kit was used to detect cell apoptosis. Western blot (WB) was used to detect the expression of CyclinD1, Caspase-3 and JNK pathway-related proteins. According to the results of HLE-B3 cell viability, 10.0 μmol/L cryptotanshinone was selected for subsequent experiments. SOD level, proliferation rate and CyclinD1 protein level in H2O2 group were lower than those in control group (P<0.05), MDA level, apoptosis rate, Caspase-3 and phosphorylation (p) -JNK protein levels were higher than those in control group (P<0.05). Cryptotanshinone group and SP 600125 group significantly inhibited the above effects of H2O2 on HLE-B3 cells (P<0.05). Compared with cryptotanshinone group, inhibitor group enhanced the effect of cryptotanshinone on H2O2-induced HLE-B3 cells (P<0.05), and activator group weakened the effect of cryptotanshinone on H2O2-induced HLE-B3 cells (P<0.05). Cryptotanshinone inhibits H2O2-induced oxidative stress and apoptosis of HLE-B3 by inhibiting JNK signaling pathway, and promotes its proliferation.
Key words: cataract; human crystalline epithelial cells; cryptotanshinone; hydrogen peroxide; C-Jun N-terminal kinase signaling pathway; oxidative stress
（Acta Laser Biology Sinica, 2025, 34(1): 090-096）