兰州大学机构库 >学院待认领
促红细胞生成素预防人脐静脉内皮细胞凋亡时对Caspase-3亚家族的影响
Alternative TitleEffect of erythropoietin on the Caspase-3 subfamily against human umbilical vein endothelial cells apoptosis
熊建文
Thesis Advisor王志禄
2009-05-26
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword促红细胞生成素 脐静脉 内皮细胞 信号通路 氧化低密度脂蛋白
Abstract目的和背景:促红细胞生成素(Erythropoietin,EPO)对心肌缺血再灌注损伤具有保护作用,但对氧化低密度脂蛋白(Oxidized low density lipoprotein, Ox-LDL)诱导的血管内皮细胞凋亡中的Caspases-3亚家族有无影响尚不清楚。我们拟探讨重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO)在预防Ox-LDL诱导人脐静脉内皮细胞(Human umbilical vein endothelial cells, HUVECs)凋亡过程中对Caspases-3亚家族的影响。 方法:体外培养HUVECs,分别加入Caspase-3抑制剂(DEVD-CHO)、Caspase-8抑制剂(z-IETD-fmk)、Caspase-9抑制剂(z-LEHD-fmk) 各25 μmol/L预孵育24h后,再加入100 μg/mL Ox-LDL处理48h,比色法检测Caspase-3,-8,-9 活性的变化,噻唑蓝(Methyl thiazolyl tetrazolium, MTT)法检测HUVECs活力。平行实验中,加入不同浓度的rhEPO预处理HUVECs 24h,再加入100 μg/mL Ox-LDL处理48h,MTT法检测HUVECs活力,流式细胞术(Flow cytometry, FCM)检测HUVECs Caspase-3蛋白阳性表达率和细胞凋亡率、比色法分别检测HUVECs Caspase-3, -8, -9酶活性。 结果:Caspase-3抑制剂组HUVECs Caspase-3活性较Ox-LDL组降低(0.80±0.17 VS 3.12±1.03,P<0.05),细胞存活率增加(74.21% ±1.68% VS 58.03%±3.21%,P<0.05); Caspase-8抑制剂组Caspase-8活性较Ox-LDL组降低(0.60±0.16 VS 1.03±0.36,P<0.05),细胞存活率无明显差异(57.24% ±4.45% VS 58.03%±3.21%,P>0.05);Caspase-9抑制剂组Caspase-3,9活性较Ox-LDL组均降低(1.02±0.21 VS 3.12±1.03;1.30±0.42 VS 3.07±0.29,P<0.05,依次),细胞存活率增加(74.08% ±3.64% VS 58.03%±3.21%,P<0.05)。6.25 IU/ml rhEPO预处理组HUVECs Caspase-3,-9活性、Caspase-3蛋白阳性表达率和细胞凋亡率较Ox-LDL组均降低(10.81±0.67 VS 12.74±1.34,3.64±0.21 VS 5.01±0.36,45.67%±6.65% VS 48.93%±3.11%,22.13%±2.16% VS 35.42%±1.32%,P<0.05,依次);25 IU/ml rhEPO预处理组HUVECs Caspase-3,-9活性、Caspase-3蛋白阳性表达率和细胞凋亡率较Ox-LDL组均降低(6.43±1.06 VS 12.74±1.34,2.03±0.37 VS 5.01±0.36,13.65% ± 4.46% VS 48.93%±3.11%,13.38%±2.35% VS 35.42%±1.32%,P<0.05,依次);100 IU/ml rhEPO预处理组人HUVECs Caspase-3,-9活性、Caspase-3蛋白阳性表达率和细胞凋亡率较Ox-LDL组均降低(3.65±0.78 VS 12.74±1.34,1.67±0.13 VS 5.01 ±0.36,3.36%±0.42% VS 48.93%±3.11%,5.77%±1.13% VS 35.42%±1.32%,P<0.05,依次);而不同浓度rhEPO预处理组HUVECs Caspase-8活性无明显变化(1.01±0.41 VS 1.06±0.23, P>0.05)。rhEPO预处理对HUVECs Caspase-3,-9活性、Caspase-3蛋白阳性表达率和细胞凋亡率的影响呈浓度依赖性(10.81±0.67 VS 6.43±1.06 VS 3.65±0.78;3.64±0.21 VS 2.03±0.37 VS 1.67±0.13;45.66%±6.65% VS 13.65%±4.46% VS 3.36%±0.42%;22.13%±2.16% VS 13.38%±2.35% VS 5.77% ± 1.13%;P<0.05,依次)。6.25、25、100 IU/ml rhEPO预处理组HUVECs 存活率...
Other AbstractObjective and Backgroud:There were many studies on the non-hematopoietic protective effect of erythropoietin on ischemia-reperfusion injury in myocardial, but the effect of erythropoietin on the Caspase-3 sub-family activity mediated by oxidized low density lipoprotein in endothelial cell is unclear. We will investigate effect of rhEPO on Caspases signaling pathway in preventing human umbilical vein endothelial cell (HUVECs) from apoptosis induced by oxidized LDL (Ox-LDL) . Methods:HUVECs were cultured in a standard medium (M 131), preincubated with inhibitors of either Caspase-3 or Caspase-8 or Caspase-9(25 μmol/L, respectively) for 24hours, then treated with Ox-LDL for 48 hours, the activities of Caspase-3,-8,-9 were measured by Caspase Colorimetric Assay Kit, cell viability was detected by methyl thiazolyl tetrazolium method. In parallel experiments, HUVECs were incubated with rhEPO (0, 6.25, 25, 100 IU/ml) for 24 hours and then treated with Ox-LDL for 48 hours, cell viability was detected by methyl thiazolyl tetrazolium method, the positive expression rate of Caspase-3 and apoptotic rate were measured by flow cytometer, the activities of Caspase-3,-8,-9 were measured by Caspase Colorimetric Assay Kit. Results:Compared with Ox-LDL group, pretreatment of Caspase-3 inhibitor decreased Caspase-3 activity (0.80 ± 0.17 VS 3.12 ± 1.03, P<0.05), increased cell survival rate (74.21%±1.68% VS 58.03%±3.21%, P<0.05 ); Caspase-8 activity of Caspase-8 inhibitor group was higher than that of Ox-LDL group(0.60± 0.16 VS 1.03 ± 0.36, P<0.05), but have no significant difference between Caspase-8 inhibitor group and Ox-LDL group in cell viability (57.24%±4.45% VS 58.03%±3.21%, P>0.05); Caspase-3, 9 activities of Caspase-9 inhibitior group were lower than that of Ox-LDL group(1.02±0.21 VS 3.12±1.03;1.3±0.42 VS 3.07±0.29, P<0.05, respectively), meanwhile, cell survival rate of Caspase-9 inhibitior group was higher (74.08%± 3.64% VS 58.03%±3.21%, P<0.05). 6.25 IU/mL rhEPO pretreatment to the culture, Caspase-3, -9 activity, positive expression rate of Caspase-3 and apoptotic rate were decreased as compared with Ox-LDL group(10.81±0.67 VS 12.74±1.34, 3.64±0.21 VS 5.01±0.36, 45.67%±6.65% VS 48.93%±3.11%, 22.10%±2.10% VS 35.40%±1.30%, P<0.05, respectively), 25 IU/mL rhEPO pretreatment to the culture, Caspase-3, -9 activity, positive expression rate of Caspase-3 and apoptotic rate were decreased as compared with Ox-LDL group (6.43±1.06 VS 12.74±1....
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223710
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
熊建文. 促红细胞生成素预防人脐静脉内皮细胞凋亡时对Caspase-3亚家族的影响[D]. 兰州. 兰州大学,2009.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Altmetrics Score
Google Scholar
Similar articles in Google Scholar
[熊建文]'s Articles
Baidu academic
Similar articles in Baidu academic
[熊建文]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[熊建文]'s Articles
Terms of Use
No data!
Social Bookmark/Share
No comment.
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.