实用医学杂志
實用醫學雜誌
실용의학잡지
THE JOURNAL OF PRACTICAL MEDICINE
2014年
15期
2381-2383
,共3页
邹秀兰%王观峰%李文立%皮荣标%俞永珍%邹玉平
鄒秀蘭%王觀峰%李文立%皮榮標%俞永珍%鄒玉平
추수란%왕관봉%리문립%피영표%유영진%추옥평
丙烯醛%ARPE-19细胞%硫辛酸烟酸二联体%Bax%Bcl-2
丙烯醛%ARPE-19細胞%硫辛痠煙痠二聯體%Bax%Bcl-2
병희철%ARPE-19세포%류신산연산이련체%Bax%Bcl-2
Acrolein%ARPE-19 cell%Lipoic acid-niacin diad%Bax%Bcl-2
目的:探讨Bax、Bcl-2蛋白在硫辛酸烟酸二联体拮抗丙烯醛诱导的体外培养 ARPE-19细胞凋亡中可能的调控作用。方法:ARPE-19细胞置于含有10%体积分数胎牛血清的DMEM培养基,37℃、体积分数5%CO2培养箱中培养,待培养瓶底部ARPE-19细胞长至70%时,种6孔板,用不含血清的DMEM培养基培养24 h ,细胞换液。实验分为3组:空白对照组、丙烯醛组和硫辛酸烟酸二联体组。细胞流式检测各组ARPE-19细胞凋亡情况,Western blot检测Bax、Bcl-2蛋白的表达水平。结果:细胞流式显示正常细胞在空白对照组中为94.8%,丙烯醛组中为60.98%,硫辛酸烟酸二联体组中为91.34%。Western Blot显示空白对照组的Bax/Bcl-2蛋白表达的比值为0.2939,丙烯醛组的 Bax/Bcl-2蛋白表达的比值为1.3892,硫辛酸烟酸二联体组的Bax/Bcl-2蛋白表达的比值为0.5558。结论:硫辛酸烟酸二联体对ARPE-19细胞的保护作用机制是通过激活抗凋亡蛋白Bcl-2来抑制凋亡蛋白Bax的表达。
目的:探討Bax、Bcl-2蛋白在硫辛痠煙痠二聯體拮抗丙烯醛誘導的體外培養 ARPE-19細胞凋亡中可能的調控作用。方法:ARPE-19細胞置于含有10%體積分數胎牛血清的DMEM培養基,37℃、體積分數5%CO2培養箱中培養,待培養瓶底部ARPE-19細胞長至70%時,種6孔闆,用不含血清的DMEM培養基培養24 h ,細胞換液。實驗分為3組:空白對照組、丙烯醛組和硫辛痠煙痠二聯體組。細胞流式檢測各組ARPE-19細胞凋亡情況,Western blot檢測Bax、Bcl-2蛋白的錶達水平。結果:細胞流式顯示正常細胞在空白對照組中為94.8%,丙烯醛組中為60.98%,硫辛痠煙痠二聯體組中為91.34%。Western Blot顯示空白對照組的Bax/Bcl-2蛋白錶達的比值為0.2939,丙烯醛組的 Bax/Bcl-2蛋白錶達的比值為1.3892,硫辛痠煙痠二聯體組的Bax/Bcl-2蛋白錶達的比值為0.5558。結論:硫辛痠煙痠二聯體對ARPE-19細胞的保護作用機製是通過激活抗凋亡蛋白Bcl-2來抑製凋亡蛋白Bax的錶達。
목적:탐토Bax、Bcl-2단백재류신산연산이련체길항병희철유도적체외배양 ARPE-19세포조망중가능적조공작용。방법:ARPE-19세포치우함유10%체적분수태우혈청적DMEM배양기,37℃、체적분수5%CO2배양상중배양,대배양병저부ARPE-19세포장지70%시,충6공판,용불함혈청적DMEM배양기배양24 h ,세포환액。실험분위3조:공백대조조、병희철조화류신산연산이련체조。세포류식검측각조ARPE-19세포조망정황,Western blot검측Bax、Bcl-2단백적표체수평。결과:세포류식현시정상세포재공백대조조중위94.8%,병희철조중위60.98%,류신산연산이련체조중위91.34%。Western Blot현시공백대조조적Bax/Bcl-2단백표체적비치위0.2939,병희철조적 Bax/Bcl-2단백표체적비치위1.3892,류신산연산이련체조적Bax/Bcl-2단백표체적비치위0.5558。결론:류신산연산이련체대ARPE-19세포적보호작용궤제시통과격활항조망단백Bcl-2래억제조망단백Bax적표체。
Objective To investigate the regulations of Bax , Bcl-2 in the protection of lipoic acid-niacin diad in acrolein-induced apoptosis in ARPE-19 cells. Methods The ARPE-19 cells were cultured in medium containing 10% fetal bovine serum , at 37 ℃ with 5% CO2. The ARPE-19 was transferred to 6-well plate after reaching to 70% confluence. After starvation for 24 h , the cells in 6-well plates were divided into three groups , including the blank control group , the acrolein treatment group with 50 μmol/L acrolein for 24 h , and the protection group with 100 μmol/L lipoic acid-niacin diad for 24 h and with the acrolein for another 24 h. The apoptotic cells were detected by flow cytometry assay , and expressions of Bcl-2 , Bax protein were detected by Western Blot assay. Results The percentages of normal healthy cells were 94.8%, 60.98%, and 91.34% in the blank control group , 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group , respectively. The ratios of Bax/Bcl-2 protein expression were 0.293 9, 1.389 2, and 0.555 8 in the blank control group, 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group, respectively. Conclusion The protective effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cell through promoting Bcl-2 expression and inhibiting Bax expression.