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作 者:宫璀璀[1] 郑乃刚[2] 张成德[3] 曹阳[1] 孟明利[1] 吴景兰[2]
机构地区:[1]中国人民解放军济南军区检验中心,河南郑州450042 [2]郑州大学分子细胞生物学研究中心,河南郑州450052 [3]中国人民解放军信息工程大学,河南郑州450075
出 处:《河南医学研究》2007年第2期108-110,共3页Henan Medical Research
摘 要:目的:探讨槲皮素对过氧化氢(H2O2)诱导NIH-3T3细胞凋亡的抑制作用。方法:体外培养小鼠成纤维细胞,取对数生长期的成纤维细胞分成四个实验组。对照组(C):仅加含有10%胎牛血清的DMEM培养基。槲皮素前保护组(Qb):先用含有50μmol/L的槲皮素培养基作用24 h,再换含有0.5 mmol/L H2O2作用细胞30 m in。槲皮素后保护组(Qa):先用0.5 mmol/L H2O2作用细胞30 m in,再换含有50μmol/L的槲皮素培养基作用24 h。H2O2实验组(H2):先用含有0.5 mmol/L H2O2作用细胞30 m in,再换仅含有10%胎牛血清的DMEM培养基作用24 h。取培养细胞提取DNA和制备1×107/m l细胞悬液的滴片,应用TUNEL和Ladder电泳检测细胞凋亡率;应用细胞免疫组化技术检测Bc l-2、Bax、Caspase-3蛋白质的表达。结果:由TUNEL和Ladder实验得出,Qb组的细胞凋亡率明显低于H2组和Qa组。由细胞免疫组化技术检测分析得出,Qb组与Qa、H2组相比,Bc l-2的表达增高,Bax、Caspase-3的表达减低。结论:槲皮素可能是通过上调Bc l-2的表达,下调Bax、Caspase-3的表达,对H2O2诱导NIH-3T3细胞的凋亡起到抑制作用。Objective: To explore effect of quercetin on antiapoptosis damage in NIH-3T3 cells. Methods : The NIH-3T3 cells in logarithmic growth phase incubated in DMEM were divided into 4 groups as follows. ①Q pre-protection group (Qb) ; prior to incubation with 0.5 mmol/L H2O2 for 30 min, the NIH-3T3 cells were incubated with 50umol/L quercetin for 24 h. ② Q post-protection group (Qa) : after incubation with 0.5 mmol/L H2O2 for 30 min, the NIH-3T3 cells were incubated with 50umoL/L quercetin for 24 h, ③ H2O2 damage group (H2) : the NIH-3T3 cells were incubated with 0, 5 mmol/L H2O2 for 30 min followed by subsequent incubation with 10% FBS for 24h. ④ The control group (C) : the NIH-3T3 cells were incubated with DMEM medium supplemented with 10% FBS. The apoptosis was detected by DNA ladder and TUNEL. The expressions of Bcl-2, Bax and Caspase-3 were demonstrated with immunocytochemistry . The NIH-3T3 cell survival rate was detected by MTT assay. Results: The apoptosis in the Qb group was decreased than that in the Qa and H2 groups. The Qb group compared with Q. and H2 groups, the expressions of Bcl-2 was increased; Bax and Caspase-3 were attenuated; and the NIH-3T3 cell survival rate was promoted. Conclusion: The anti-apoptosis effect of quercetin damage in the NIH-3T3 cells may act through down-regulated expressions of Bax and Caspase-3 and up-regulated expression of Bcl-2.
关 键 词:槲皮素 凋亡 NIH-3T3细胞系
分 类 号:R334[医药卫生—人体生理学]
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