| 陈星,周力量,刘东敬,等.微小 RNA-595在人晶状体上皮细胞凋亡中的调控作用[J].安徽医药,2021,25(11):2232-2236. |
| 微小 RNA-595在人晶状体上皮细胞凋亡中的调控作用 |
| The regulatary effect of miR-595 on apoptosis of human lens epithelial cells and its mechanism |
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| DOI：10.3969/j.issn.1009-6469.2021.11.025 |
| 中文关键词: 白内障 微小 RNA-595 细胞凋亡 B细胞淋巴瘤 -2 |
| 英文关键词: Cataract MiR-595 Apoptosis Bcl-2 |
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| 中文摘要: |
| 目的探讨微小 RNA-595(miR-595)在年龄相关性白内障晶状体组织中的表达情况及其对人晶状体上皮细胞凋亡的影响与机制。方法收集年龄相关性白内障病人晶状体前囊膜组织(白内障组)及正常供体眼球晶状体前囊膜组织(健康组)标本，以 H2O2诱导构建人晶状体上皮细胞 SRA01/04凋亡模型( H2O2组)并以正常培养的 SRA01/04细胞作为对照(对照组)，采用实时荧光定量 PCR检测 miR-595的表达改变。将体外培养的 SRA01/04，细胞分为 mimics-NC组、 miR-595 mimics组、 inhibitorNC组和 miR-595 inhibitor组，将 miR-595 mimics/inhibitor及其阴性对照 mimics/inhibitor-NC转染至 SRA01/04细胞后，采用实时荧光定量 PCR检测检测各组细胞中 miR-595的表达情况；转染 48 h后暴露于 200 μmol/L H2O2 1 h，采用流式细胞仪检测各组细胞凋亡率，免疫印迹法检测细胞中 B细胞淋巴瘤 -2(Bcl-2)蛋白的表达水平；采用双荧光素酶报告基因实验检测 miR-595和 Bcl2的靶向关系。结果与健康组相比，白内障组中 miR-595表达水平明显升高［(5.52±1.64)比( 1.00±0.00)P<0.05］；同时， H2O2组细胞中 miR-595表达水平明显高于对照组［(3.86±1.12)比( 1.00±0.00)，P<0.05］。与 mimics-NC组比较，m，iR-595 mimics组细胞中 miR-595表达水平［(4.16±0.78)比( 1.00±0.00)］、细胞凋亡率［(17.52±3.05)%比(8.86±1.21)%］明显升高，而 Bcl-2蛋白的表达水平明显降低( P<0.05)；但 miR-595 inhibitor组细胞中 miR-595表达水平、细胞凋亡率明显低于 inhibitor-NC组，而 Bcl-2蛋白的表达水平明显高于 inhibitor-NC组( P<0.05)。 Bcl-2是 miR-595的靶基因。结论 miR-595在年龄相关性白内障晶状体组织中高表达，并靶向调控 Bcl-2表达促进人晶状体上皮细胞凋亡。 |
| 英文摘要: |
| Objective To investigate the expression of miR-595 in the lens of age-related cataract and its effect on the apoptosis of human lens epithelial cells.Methods The anterior capsular tissues of age-related cataract patients (cataract group) and normal donors(normal group) were collected, the apoptosis model of human lens epithelial cells SRA01/04 (H2O2 group) was established by H2O2 induction, and the normal cultured SRA01/04 cells were used as the control group, and the expression of miR-595 was detected by real-time fluorescent quantitative PCR. SRA01/04 cells cultured in vitro were divided into mimics-NC group, miR-595 mimics group, inhibitor-NC group and miR-595 inhibitor group. After miR-595 mimics/inhibitor and its negative control mimics/inhibitor-NC were transfected into SRA01/04 cells, the expression of miR-595 in the cells of each group was detected by real-time fluorescent quantitative PCR. After transfected for 48h, and exposed to 200 μmol/L H2O2for 1h, the apoptosis rateof each groupwas detected by flow cytometry,the expression of Bcl-2 protein was detected by Western blotting, and the targeting relationship between miR-595 and Bcl-2 was detected by double luciferase reporter gene assay.Results Compared with the normal group, the expression level of miR-595 in cataract group was significantly higher [(5.52±1.64) vs. (1.00±0.00), P<0.05]; meanwhile, the expression level of miR-595 in H2O2 group was significantly higher than that in the control group[(3.86±1.12) vs. (1.00±0.00), P<0.05]. Compared with mimics-NC group, miR-595 expression level [(4.16±0.78) vs. (1.00±0.00)] and apoptosis rate [(17.52±3.05) % vs. (8.86±1.21) %] in miR-595 mimics group were significantly increased, while Bcl-2 protein expression level was significantly decreased (P<0.05), but miR-595 expression level and apoptosis rate in miR-595 inhibitor group were significantly lower than those in inhibitor NC group, while Bcl-2 protein expression level was significantly higher than that in inhibitor NC group (P<0.05). Bcl-2 was the target gene of miR-595.Conclusion miR-595 is highly expressed in the lens of age-related cataract and can promote the apoptosis of human lens epithelial cells, and the mechanism may be related to the targeted regulation of Bcl-2 expression. |
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