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                   WANG Tao,HE Jin,LIU Ya,et al.Inhibitory effect of curcumin on ultraviolet-B-induced DNA oxidative damage and apoptosis in human retinal pigment epithelial cells[J].Recent Advances in Ophthalmology,2023,43(7):530-535.[doi:10.13389/j.cnki.rao.2023.0107]





                  Inhibitory effect of curcumin on ultraviolet-B-induced DNA oxidative damage and apoptosis in human retinal pigment epithelial cells
                  215000 江蘇省蘇州市,蘇州科技城醫院眼科
                  WANG TaoHE JinLIU YaLU PingZHANG MengyuLIU Jianjun
                  Department of Ophthalmology,Suzhou Science and Technology Town Hospital,Suzhou 215000,Jiangsu Provience,China
                  curcumin ultraviolet-B human retinal pigment epithelial cell DNA oxidative damage apoptosis
                  目的 探討姜黃素在紫外線B(UVB)誘導的人視網膜色素上皮(RPE)細胞DNA氧化損傷和凋亡中的作用機制。
                  方法 將人視網膜色素上皮細胞株(ARPE-19)隨機分為空白對照組、二甲基亞砜(DMSO)組、姜黃素組、UVB照射組、UVB+DMSO組和UVB+姜黃素組。CCK-8實驗檢測姜黃素對UVB照射前后ARPE-19細胞活力的影響;免疫熒光檢測DNA氧化損傷標志蛋白磷酸化組蛋白H2AX(γH2AX)的定位;Western blot檢測γH2AX、凋亡相關蛋白[B淋巴細胞瘤-2基因(BCL-2)和BCL-2相關X蛋白(BAX)]的表達變化;使用X-rosamine衍生物(Mito-Tracker Red CMXRos)和異硫氰酸熒光素(FITC)標記的鈣離子依賴的磷脂結核蛋白Annexin V(Annexin V-FITC)來檢測UVB誘導的ARPE-19細胞線粒體膜電位變化和細胞凋亡情況。
                  結果 與空白對照組相比,DMSO組DMSO處理24 h后的ARPE-19細胞活力無明顯改變(P>0.05)。與DMSO組相比,姜黃素組采用15 μmol·L-1 姜黃素處理24 h后ARPE-19細胞活力輕度升高(P<0.05),表明15 μmol·L-1姜黃素對ARPE-19細胞無毒性作用,后續實驗采用該濃度處理細胞。與空白對照組相比,UVB照射組ARPE-19細胞活力降低,γH2AX和BAX蛋白表達水平升高,BCL-2蛋白表達水平降低,線粒體膜電位降低,細胞凋亡增加(均為P<0.05)。與UVB照射組相比,UVB+DMSO組ARPE-19細胞活力,γH2AX、BAX以及BCL-2蛋白表達水平均無明顯變化(均為P>0.05),線粒體膜電位和細胞凋亡均無明顯改變(均為P>0.05)。與UVB+DMSO組相比,UVB+姜黃素組ARPE-19細胞活力升高,γH2AX和BAX蛋白表達水平降低,BCL-2蛋白表達水平升高,線粒體膜電位升高,細胞凋亡減少(均為P<0.05)。
                  結論 姜黃素能減輕UVB誘導的RPE細胞DNA氧化損傷和凋亡,有望為姜黃素在年齡相關性黃斑變性中的作用機制研究提供新的思路。
                  Objective To investigate the effects of curcumin on DNA oxidative damage and apoptosis in human retinal pigment epithelial (RPE) cells induced by ultraviolet-B (UVB).
                  Methods Human RPE cell lines (ARPE-19) were divided into the blank control (BC) group,dimethyl sulfoxide (DMSO) group,curcumin group,UVB irradiation group,UVB+DMSO group,and UVB+curcumin group.The effect of curcumin on the activity of ARPE-19 cells before and after UVB irradiation was assessed by CCK-8 assay.The H2AX phosphorylation (γH2AX),a marker of DNA oxidative damage,was localized by immunofluorescence.The expression levels of γH2AX and apoptosis-related proteins,including B-cell lymphoma 2 (BCL-2) and Bcl-2-associated X protein (BAX),were measured by Western blot.X-rosamine derivative Mito-Tracker Red CMXRos and Annexin V labeled by fluorescein isothiocyanate (Annexin V-FITC) were used to detect UVB-induced mitochondrial membrane potential changes and apoptosis in ARPE-19 cells.
                  Results Compared with the BC group,ARPE-19 cells in the DMSO group showed no significant changes in viability after being treated with DMSO for 24 h (P>0.05).Compared with the DMSO group,the viability of ARPE-19 cells after being treated with 15 μmol·L-1 curcumin for 24 h in the curcumin group increased slightly (P<0.05),indicating that curcumin at this concentration had no cytotoxicity,and it was used in the subsequent experiments.Compared with the BC group,ARPE-19 cell viability,BCL-2 level,and mitochondrial membrane potential in the UVB irradiation group decreased,while γH2AX and BAX levels and cell apoptosis increased (all P<0.05).Compared with the UVB irradiation group,ARPE-19 cell viability,γH2AX,BAX and BCL-2 levels,mitochondrial membrane potential,and cell apoptosis in the UVB+DMSO group showed no significant changes (all P>0.05).Compared with the UVB+DMSO group,ARPE-19 cell viability,BCL-2 level,and mitochondrial membrane potential increased,while γH2AX and BAX levels and cell apoptosis decreased in the UVB+curcumin group (all P<0.05).
                  Conclusion Curcumin can reduce DNA oxidative damage and apoptosis in RPE cells induced by UVB,which provides a new idea for studying the effects of curcumin on age-related macular degeneration.


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                  更新日期/Last Update: 2023-07-05

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