rHDL抑制CACs衰老的机制研究

发布时间：2018-05-19 17:10

  本文选题：rHDL + CACs衰老　； 参考：《北京协和医学院》2011年硕士论文

【摘要】：目的 循环成血管细胞(circulating angiogenic cells, CACs)参与新生血管的形成和动脉修复。高密度脂蛋白(HDL)可提高CACs的活性,但其中的调节机制迄今仍不清楚。本研究旨在阐述rHDL是通过什么分子机制来影响CACs衰老的。方法 1、从健康志愿者外周血中分离单个核细胞,并诱导分化培养为CACs。 2、CACs与rHDL共培养后(加或不加雷帕霉素),用β-半乳糖苷酶染色法检测CACs衰老程度。 3、Western Blot口免疫沉淀法检测：nTOR磷酸化、(?)nTOR-rictor复合物形成和mTOR-rictor依赖的Akt活化。 4、免疫细胞组织化学法、RT-PCR及Western Blot检测人端粒酶逆转录酶(hTERT)的核转位,试剂盒检测核端粒酶活性。结果 1、rHDL可减弱CACs的衰老,然而此效应可被哺乳动物雷帕霉素靶蛋白(mTOR)抑制剂雷帕霉素所阻断。 2、rHDL能促使mTOR发生磷酸化、mTOR-rictor复合物形成和mTOR-rictor依赖的Akt活化；促使人端粒酶逆转录酶(hTERT)的核转位增加及核端粒酶活性增强。 3、用SiRNA抑制rictor基因表达后,(?)nTOR-rictor复合物形成和Akt活化均被阻断,同时rHDL对CACs衰老的抑制作用和核端粒酶活性的提高作用均被抑制。结论 rHDL能促使(?)nTOR的持久磷酸化和(?)nTOR-rictor复合物形成,并能通过mTORC2信号通路的激活而抑韦CACs的衰老。
[Abstract]:Purpose Circulating angiogenic cells, CACs) is involved in angiogenesis and arterial repair. High density lipoprotein (HDL) can improve the activity of CACs, but the regulatory mechanism is still unclear. The aim of this study was to elucidate the molecular mechanism by which rHDL affects CACs senescence. Method 1. Mononuclear cells were isolated from the peripheral blood of healthy volunteers and differentiated into CACs. (2) after co-culture with rHDL (with or without rapamycin), 尾 -galactosidase staining was used to detect the senescence of CACs. 3Western Blot immunoprecipitation assay was used to detect the formation of nTOR-rictor complex and the activation of mTOR-rictor dependent Akt. 4. The nuclear translocation of human telomerase reverse transcriptase (hTERT) was detected by RT-PCR and Western Blot, and the nuclear telomerase activity was detected by kit. Result 1rHDL can attenuate the senescence of CACs, but this effect can be blocked by rapamycin, an inhibitor of rapamycin target protein mTORin in mammals. 2rHDL could induce the formation of phosphorylated mTOR-rictor complex and the activation of mTOR-rictor dependent Akt, and increase the nuclear translocation and nuclear telomerase activity of human telomerase reverse transcriptase (hTERT). 3. The inhibition of rictor gene expression by SiRNA and the inhibition of rHDL on CACs senescence and the increase of nuclear telomerase activity were all blocked. Conclusion RHDL can promote the sustained phosphorylation of nTOR and the formation of nTOR-rictor complex, and inhibit the senescence of CACs through the activation of mTORC2 signaling pathway.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

【参考文献】

相关期刊论文 前4条

1 尹扬光;黄岚;赵晓辉;于世勇;方玉强;赵景红;崔斌;;基质细胞衍生因子1α介导小鼠内皮祖细胞修复损伤血管内膜[J];中国动脉硬化杂志;2007年01期

2 童中艺;王佐;姜志胜;宋砚明;周晓峰;田永凤;;基质细胞衍生因子1α对大鼠骨髓源内皮祖细胞迁移的影响[J];中国动脉硬化杂志;2007年07期

3 陈剑飞;黄岚;晋军;武晓静;赵晓晖;崔斌;于世勇;赵刚;;年龄对骨髓内皮祖细胞功能的影响及意义[J];四川医学;2006年05期

4 朱军慧,陈君柱,王兴祥,朱建华,尚云鹏,郭晓纲,戴禾敏;氧化低密度脂蛋白对外周血内皮祖细胞数量和功能的影响[J];中华内分泌代谢杂志;2005年01期

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