学号:
泰山医学院毕业设计（论文）
题目： 蜂胶黄酮抗H2O2诱导PC12细胞
凋亡机制的研究
院（部）系 所 学 专 业 年级、班级 完成人姓名 指导教师姓名 专业技术职称
药学院 药学
2013年 6 月 18 日
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专业： 班级： 签名： 20 年 月 日
摘 要
目的 观察蜂胶黄酮对过氧化氢（H2O2）诱导大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡的影响及机制。
方法 培养PC12细胞，取对数生长期细胞分为五组，空白对照组、模型组、蜂胶黄酮高、中、低剂量组，剂量分别为200mg/L、100 mg/L、50 mg/L.药物预处理2h后，孵育H2O2（140μmol/L）24h诱导过氧化损伤。TUNEL试剂盒检测原位细胞凋亡,流式细胞仪检测细胞内活性氧水平以及细胞周期，ELISA检测细胞内Caspase-3蛋白含量。
结果 TUNEL细胞凋亡染色显示, H2O2组与空白对照组比较染色明显加深,而蜂胶黄酮组染色明显变浅，说明蜂胶黄酮能对抗H2O2诱导细胞凋亡；周期结果显示H2O2组处于G0/G1细胞明显增多，而处于G2/M、S期细胞明显减少，细胞增殖降低，而蜂胶黄酮增加S期细胞促进细胞增殖；与空白对照组相比H2O2组活性氧水平、细胞内Caspase-3含量明显增高，而蜂胶黄酮各剂量组活性氧水平降低，Caspase-3含量减少。
结论 蜂胶黄酮对H2O2诱发PC12神经细胞凋亡有显着的抑制作用，其机制可能其影响细胞周期、清除氧自由基、降低凋亡因子Caspase-3有关。
关键词 蜂胶黄酮；PC12细胞；H2O2;Caspase-3；细胞凋亡
Abstract
Objective:?To observe the protection of?propolis flavonoids?on rat with injuried pheochromocytomacells(PC12) induced by??hydrogen peroxide(H2O2)?and to explore its possible mechanism.
Methods:???Culture?PC12 cells,the cells in?the logarithmic growth phase were?divided into five groups,blank control group,model group,propolis flavone?of high,medium and low dose group,the dose of 200mg/L,100 mg/L,50 mg/L.?After 2h 0f?Pharmacological preconditioning,the cells?were incubated with H2O2(140 μ mol/L)?for?24h?to induce?oxidative damage.?TUNEL Kit?is used for?determining Cell apoptosis,Flow cytometry was used to detect the intracellular ROS level and cell cycle,ELISA is used for determining the concentration of protein Caspase-3 in cells.
Results:?Apoptosis of TUNEL cells staining, H2O2 group compared with the control group, staining was deepened, and the propolis flavone group was significantly lighter, that propolis flavonoids can antagonize the apoptosis induced by H2O2 Cycle showed that H2O2 group in G0/G1 cells were increased, and in the G2/M,S phase cells decreased significantly, reduced cell proliferation, and propolis flavonoids increased S phase cells promoting cell proliferation; compared with the blank control group, caspase-3 in H2O2group, the levels of reactive oxygen species in cells increased significantly, while the propolis flavone in all dose groups decrease the levels of reactive oxygen species, reduced caspase-3 content.
Conclusion: Propolis flavonoids on H2O2-induced PC12 cells damage a significant protective effect,The mechanism may be its effect on cell cycle, scavenging oxygen free radicals, decrease the apoptosis related factor caspase-3.
Key words: Propolis; flavonoids; PC12 cells; H2O2; apoptosis
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