| 促红细胞生成素对TNF-α诱导的血管内皮细胞凋亡的影响 |
| Alternative Title | Effects of erythropoietin on vascular endothelial cells apoptosis induced by tumor necrosis factor-alpha
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| 李侃 |
| Thesis Advisor | 李建华
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| 2012-05-25
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| Degree Grantor | 兰州大学
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| Place of Conferral | 兰州
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| Degree Name | 硕士
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| Keyword | 促红细胞生成素
慢性肾脏病
肿瘤坏死因子-α
血管内皮细胞
内皮型一氧化氮合酶
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| Abstract | 目的：本研究通过观察rhEPO对TNF-α诱导的HUVEC凋亡的影响，评价EPO对eNOS的积极作用是否同样也发生于血管内皮细胞以及EPO对细胞凋亡相关的胞内信号传导元件Caspase-3表达的影响。以期为EPO替代治疗在降低CVD发生率和延缓CKD进展中的应用提供理论依据。方法：体外培养的HUVEC随机分为5组：正常对照组、TNF-α组、rhEPO低、中、高剂量组。倒置荧光显微镜观察细胞生长形态，MTT法检测细胞生存率，SCGE法（彗星实验）检测细胞凋亡率，ELISA法检测eNOS的浓度，免疫细胞化学法检测Caspase-3蛋白的表达。结果：TNF-α作用24h后HUVEC呈现出胞体皱缩、胞膜发泡和胞核致密等凋亡形态学变化。与正常对照组相比，TNF-α组细胞生存率显著降低，而细胞凋亡率显著升高，凋亡诱导效应的产生伴随eNOS浓度的降低和Caspase-3相对含量的升高（P均<0.01）。与TNF-α组相比，rhEPO预处理各组细胞形态更趋近于正常；中、高剂量rhEPO可以提高eNOS的浓度（P均<0.05），而低剂量rhEPO对eNOS的浓度没有产生显著的影响（P>0.05）；与此同时，rhEPO剂量依赖性地降低Caspase-3蛋白的相对含量，提高细胞生存率，降低细胞凋亡率（P均<0.05）。rhEPO预处理各组组间比较差异有统计学意义（P均<0.05）。结论：EPO可以抑制TNF-α诱导的血管内皮细胞凋亡，抗凋亡机制可能与其恢复eNOS的活化和抑制Caspase-3的表达有关。 |
| Other Abstract | Objective: We investigated the effects of rhEPO on HUVEC apoptosis induced by TNF-α. Methods: Cultured endothelial cells were randomly divided into five study groups. Morphology of endothelial cells was observed by inverted fluorescence microscope. Cell viability was measured by MTT assay. Cell apoptosis was evaluated using SCGE assay. The concentration of eNOS was assessed using ELISA. The expression of Caspase-3 protein was examined by immunocytochemistry assay. Results: TNF-α-treated HUVEC displayed morphological features of apoptosis. TNF-α-induced endothelial cell apoptosis was associated with reduced eNOS concentration but increased Caspase-3 protein expression. rhEPO-pretreated HUVEC exhibited better morphology. Pretreatment with rhEPO at 25 and 50 IU/ml restored eNOS activation. This phenomenon was accompanied by reduced the relative content of Caspase-3 protein, as resulting in improved cell viability and reduced apoptosis in a dose-dependent manner. Conclusion: The anti-apoptotic effects of EPO on vascular endothelial cells are mediated by restoring eNOS activation and inhibiting Caspase-3 expression. |
| URL | 查看原文
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| Language | 中文
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| Document Type | 学位论文
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| Identifier | https://ir.lzu.edu.cn/handle/262010/223714
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| Collection | 学院待认领
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| Affiliation | 临床医学院
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Recommended Citation
GB/T 7714 |
李侃. 促红细胞生成素对TNF-α诱导的血管内皮细胞凋亡的影响[D]. 兰州. 兰州大学,2012.
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