槲皮素抗NIH-3T3细胞凋亡的作用

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。

Anti-apoptosis effect of quercetin on damage in NIH-3T3 cells

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.① Q pre-protection group(Qb);prior to incubation with 0.5 mmol/LH2O2 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 Qa 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.