槲皮素抗NIH-3T3细胞凋亡的作用

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

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.