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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）系药学院

所学专业药学

年级、班级

完成人姓名

指导教师姓名

专业技术职称

2013年 6 月 18 日

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20 年月日

摘要

目的观察蜂胶黄酮对过氧化氢（H

2O

2

）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H

2O

2

（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, H

2O

2

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H

2O

2

诱导细胞凋亡；周期结果显示H

2

O

2

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H

2O

2

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H

2O

2

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；H

2O

2

;Caspase-3；细胞凋亡

Abstract

Objective:

To observe the protection ofpropolis flavonoidson rat with injuried

pheochromocytomacells(PC

12) induced by?hydrogen peroxide(H

2

O

2

)

and to explore its possible mechanism.

Methods:Culture?PC

12

cells,the cells inthe logarithmic growth phase weredivided into five groups,blank control group,model group,propolis flavoneof

high,medium and low dose group,the dose of 200mg/L,100 mg/L,50 mg/L.After 2h

0fPharmacological preconditioning,the cellswere incubated with H

2O

2

(140 μ

mol/L)for24hto induceoxidative damage.TUNEL Kitis used fordetermining 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, H

2O

2

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 H

2O 2

Cycle showed that H

2O

2

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 H

2O

2

group, 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 H

2O

2

-induced PC

12

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; PC

12 cells; H

2

O

2

; apoptosis