[1]徐丽珲,郭玉,伏等弟,等.紫外线照射对人视网膜色素上皮细胞活性和沉默信息调节因子6（SIRT6)表达的影响[J].眼科新进展,2021,41(9):817-821.[doi:10.13389/j.cnki.rao.2021.0171]
　XU Lihui,GUO Yu,FU Dengdi,et al.Effect of ultraviolet radiation on the activity of human retinal pigment epithelial cells and the expression level of silent information regulator 6[J].Recent Advances in Ophthalmology,2021,41(9):817-821.[doi:10.13389/j.cnki.rao.2021.0171]
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紫外线照射对人视网膜色素上皮细胞活性和沉默信息调节因子6（SIRT6)表达的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
41卷
期数:
2021年9期
页码:
817-821
栏目:
实验研究
出版日期:
2021-09-05

文章信息/Info

Title:
Effect of ultraviolet radiation on the activity of human retinal pigment epithelial cells and the expression level of silent information regulator 6
作者:
徐丽珲; 郭玉; 伏等弟; 蔡莲君; 芦晓红; 罗明秀; 姚青
750004　宁夏回族自治区银川市，宁夏医科大学基础医学院生物化学与分子生物学系（徐丽珲，伏等弟，蔡莲君，芦晓红，罗明秀，姚青）；750004　宁夏回族自治区银川市，宁夏医科大学基础医学院生物技术专业（郭玉）
Author(s):
XU Lihui1; GUO Yu2; FU Dengdi1; CAI Lianjun1; LU Xiaohong1; LUO Mingxiu1; YAO Qing1
1.Department of Biochemistry and Molecular Biology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Autonomous Region,China
2.Department of Biotechnology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Autonomous Region,China
关键词:
紫外线; 人视网膜色素上皮细胞; 沉默信息调节因子6
Keywords:
ultraviolet;  human retinal pigment epithelial cells;  silent information regulator 6
分类号:
R774
DOI:
10.13389/j.cnki.rao.2021.0171
文献标志码:
A
摘要:
目的　探讨紫外线照射对人视网膜色素上皮细胞（ARPE-19细胞）活性和沉默信息调节因子6（SIRT6)表达的影响。方法　将对数生长期的ARPE-19细胞按照照射剂量随机分为对照组（0 mJ·cm-2）及7.5 mJ·cm-2、15.0 mJ·cm-2、30.0 mJ·cm-2、60.0 mJ·cm-2、120.0 mJ·cm-2、240.0 mJ·cm-2照射剂量组进行干预培养。照射后继续常规培养24 h，采用倒置荧光显微镜观察细胞形态并拍照，CCK-8法检测细胞存活率，Western blot检测各组细胞中SIRT6的表达情况，采用免疫荧光染色对对照组、30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组ARPE-19细胞中SIRT6进行定位检测。结果　对照组ARPE-19细胞呈梭形贴壁生长，边界清楚，连接紧密，细胞状态良好；30.0 mJ·cm-2照射剂量组ARPE-19细胞形态不规整，细胞数量略有减少，细胞间连接疏松。CCK-8法检测结果显示：与对照组相比，ARPE-19细胞在不同剂量紫外线照射后，细胞存活率显著降低且呈剂量依赖性（均为P＜0.05）。与对照组相比，除较低照射剂量组（7.5 mJ·cm-2、15.0 mJ·cm-2）外，其余各组SIRT6蛋白相对表达量均降低，且随着照射剂量的增加呈剂量依赖性逐渐降低（均为P＜0.05）。经过30.0 mJ·cm-2紫外线照射后，ARPE-19细胞随着再培养时间的延长，细胞中SIRT6蛋白的相对表达量逐渐下降（均为P＜0.05）。对照组、30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组SIRT6主要表达于ARPE-19细胞核内；与对照组相比，30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组ARPE-19细胞中SIRT6的荧光强度显著降低，且240.0 mJ·cm-2照射剂量组降低更明显。结论　紫外线照射能够降低ARPE-19细胞的增殖活性，同时也能下调细胞中SIRT6的表达。
Abstract:
Objective　To observe the effects of the ultraviolet radiation on the activity of human retinal pigment epithelial cells (ARPR-19) and the expression level of silent signal regulatory protein 6 (SIRT6).Methods　ARPE-19 cells in the logarithmic phase were randomly divided into seven groups and treated with ultraviolet radiation at the doses of 0 mJ·cm-2 (control group), 7.5 mJ·cm-2, 15.0 mJ·cm-2, 30.0 mJ·cm-2, 60.0 mJ·cm-2, 120.0 mJ·cm-2 and 240.0 mJ·cm-2, respectively. After 24 h radiation, the cell morphology was observed by the inverted microscope. CCK-8 was used to detect cell activity. Protein level of SIRT6 was detected by Western blot. Immunofluorescence staining was used to detect the localization of SIRT6 in ARPE-19 cells in control group, 30.0 mJ·cm-2 group and 240.0 mJ·cm-2 group.Results　In the control group, fusiform-like ARPE-19 cells adherently grew with a clear boundary, tight connection and good cell condition. In the 30.0 mJ·cm-2 group, ARPE-19 cells presented an irregular morphology and loose cell-cell connection, and the number of cells decreased slightly. CCK-8 results showed that compared with the control group, cell survival rate of ARPE-19 cells significantly decreased in a dose-dependent manner after different doses of ultraviolet radiation (P<0.05). Compared with the control group, protein level of SIRT6 dose-dependently downregulated in all groups except for the lower dose radiation at 7.5 mJ·cm-2 and 15.0 mJ·cm-2 (P<0.05). After 30.0 mJ·cm-2 ultraviolet radiation, protein level of SIRT6 protein in ARPE-19 cells time-dependently downregulated with the prolongation of re-culture time (P<0.05). SIRT6 was mainly expressed in the nucleus of ARPE-19 cells in the control group, 30.0 mJ·cm-2 group and 240.0 mJ·cm-2 group. The fluorescence intensity of SIRT6 in ARPE-19 cells of 30.0 mJ·cm-2 group and 240.0 mJ·cm-2 group was significantly lower than that in control group, especially in 240.0 mJ·cm-2 group.Conclusion　Ultraviolet radiation can reduce the proliferative activity of ARPE-19 cells and downregulate the protein level of SIRT6.

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备注/Memo

备注/Memo:
国家自然科学基金资助（编号：82060819）；宁夏回族自治区自然科学基金资助（编号：2020AAC02024）；宁夏医科大学校级科研项目（编号：XM2020012）
更新日期/Last Update: 2021-09-05