中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
10期
1839-1844
,共6页
王雯婕%陈剑%刘小勇%曲艺欣%周清%袁晟辰%郝晓宁
王雯婕%陳劍%劉小勇%麯藝訢%週清%袁晟辰%郝曉寧
왕문첩%진검%류소용%곡예흔%주청%원성신%학효저
白藜芦醇%ARPE-19细胞%细胞周期%细胞增殖
白藜蘆醇%ARPE-19細胞%細胞週期%細胞增殖
백려호순%ARPE-19세포%세포주기%세포증식
Resveratrol%ARPE-19 cells%Cell cycle%Cell proliferation
目的：探讨白藜芦醇（ resveratrol，Res）对视网膜色素上皮细胞增殖的影响，并初步探讨其作用机制。方法：用0、50、100、150、200和300μmol/L Res作用于ARPE-19细胞24、48和72 h后，CCK-8法检测Res对ARPE-19细胞增殖的影响，0、100、150和200μmol/L Res作用ARPE-19细胞48 h，PI单染流式细胞术检测细胞周期，Annexin V-FITC/PI双染流式细胞术检测细胞凋亡，免疫荧光化学法检测PCNA蛋白表达，实时荧光定量PCR （real-time PCR）法检测PCNA、P21和P27的mRNA表达水平。结果： CCK-8结果显示，Res抑制ARPE-19细胞增殖，呈时间和剂量依赖性；流式细胞术显示，Res使S期细胞百分比显著上升且各组凋亡率没有区别；免疫荧光显示， Res抑制PCNA蛋白表达，荧光减弱；real-time PCR显示， Res浓度依赖性地抑制PCNA的mRNA表达，而诱导表达P21和P27的mRNA。结论：Res能抑制ARPE-19细胞的增殖，使细胞阻滞在S期，其机制可能与白藜芦醇诱导P21与P27的mRNA表达、抑制PCNA的mRNA表达有关。
目的：探討白藜蘆醇（ resveratrol，Res）對視網膜色素上皮細胞增殖的影響，併初步探討其作用機製。方法：用0、50、100、150、200和300μmol/L Res作用于ARPE-19細胞24、48和72 h後，CCK-8法檢測Res對ARPE-19細胞增殖的影響，0、100、150和200μmol/L Res作用ARPE-19細胞48 h，PI單染流式細胞術檢測細胞週期，Annexin V-FITC/PI雙染流式細胞術檢測細胞凋亡，免疫熒光化學法檢測PCNA蛋白錶達，實時熒光定量PCR （real-time PCR）法檢測PCNA、P21和P27的mRNA錶達水平。結果： CCK-8結果顯示，Res抑製ARPE-19細胞增殖，呈時間和劑量依賴性；流式細胞術顯示，Res使S期細胞百分比顯著上升且各組凋亡率沒有區彆；免疫熒光顯示， Res抑製PCNA蛋白錶達，熒光減弱；real-time PCR顯示， Res濃度依賴性地抑製PCNA的mRNA錶達，而誘導錶達P21和P27的mRNA。結論：Res能抑製ARPE-19細胞的增殖，使細胞阻滯在S期，其機製可能與白藜蘆醇誘導P21與P27的mRNA錶達、抑製PCNA的mRNA錶達有關。
목적：탐토백려호순（ resveratrol，Res）대시망막색소상피세포증식적영향，병초보탐토기작용궤제。방법：용0、50、100、150、200화300μmol/L Res작용우ARPE-19세포24、48화72 h후，CCK-8법검측Res대ARPE-19세포증식적영향，0、100、150화200μmol/L Res작용ARPE-19세포48 h，PI단염류식세포술검측세포주기，Annexin V-FITC/PI쌍염류식세포술검측세포조망，면역형광화학법검측PCNA단백표체，실시형광정량PCR （real-time PCR）법검측PCNA、P21화P27적mRNA표체수평。결과： CCK-8결과현시，Res억제ARPE-19세포증식，정시간화제량의뢰성；류식세포술현시，Res사S기세포백분비현저상승차각조조망솔몰유구별；면역형광현시， Res억제PCNA단백표체，형광감약；real-time PCR현시， Res농도의뢰성지억제PCNA적mRNA표체，이유도표체P21화P27적mRNA。결론：Res능억제ARPE-19세포적증식，사세포조체재S기，기궤제가능여백려호순유도P21여P27적mRNA표체、억제PCNA적mRNA표체유관。
AIM:To investigate the effects of resveratrol ( Res) on the proliferation of ARPE-19 cells and to ex-plore the possible mechanisms.METHODS:After ARPE-19 cells were treated with Res at concentrations of 0, 50, 100, 150, 200 and 300 μmol/L for 24 h, 48 h and 72 h, the effects of Res on the proliferation of the cells were tested by CCK-8 assay.The ARPE-19 cells were treated with Res at concentrations of 0, 100, 150 and 200 μmol/L for 48 h.The effects of Res on the cell cycle and apoptosis were detected by flow cytometry with Annexin V-FITC/PI staining.The protein expression of proliferating cell nuclear antigen (PCNA) was detected by immunofluorescent assay.The mRNA expression of PCNA, P21 and P27 was determined by real-time PCR.RESULTS:The results of CCK-8 assay showed that Res inhibited the prolifera-tion of ARPE-19 cells in a time-and dose-dependent manner.The treatment with Res for 48 h resulted in an arrest of cell cycle at S phase without increasing cell apoptosis.Res inhibited the protein expression of PCNA in ARPE-19 cells.The re-sults of real-time PCR showed that Res increased the mRNA expression of P21 and P27, and decreased the mRNA expres-sion of PCNA.CONCLUSION: Res inhibits the proliferation of ARPE-19 cells and induces the cell cycle arrest at S phase.The mechanism may be related to up-regulation of P21 and P27, and down-regulation of PCNA.
