内皮源性降钙素基因相关肽抗血管紧张素Ⅱ诱导的内皮细胞凋亡作用

发布时间：2018-05-27 07:16

  本文选题：内皮源性降钙素基因相关肽 + 人脐静脉内皮细胞　； 参考：《中国现代医学杂志》2013年24期

【摘要】：目的探讨内皮源性降钙素基因相关肽对血管紧张素II诱导的内皮细胞抗凋亡作用。方法培养HUVEC-12,分别加入不同剂量的AngII孵育内皮细胞,研究其量效和时效关系,并给予特异性AT1受体阻断剂氯沙坦研究其可能的机制。Real time-PCR检测内皮细胞中CGRP mRNA水平;放射免疫法检测内皮细胞CGRP的含量。在培养HUVEC-12给予AngII诱导细胞凋亡,外源性给予CGRP和辣椒素。Annexin V-FITC流式细胞检测细胞凋亡;比色法测定caspase-3活性;RT-PCR检测内皮细胞中CGRP、Bcl-2和Bax mRNA的表达。结果①AngII孵育内皮细胞可剂量和时间依赖性地降低α-和β-CGRP mRNA的表达水平和CGRP含量,此作用可被预先给予的氯沙坦阻断。②CGRP可剂量依赖性降低AngII诱导的内皮细胞凋亡,减弱AngII增加内皮细胞caspase-3活性,此作用能被预先给予的CGRP受体阻断剂CGRP8-37取消。③辣椒素可剂量依赖性上调内皮源性α-和β-CGRP mRNA的表达水平,此作用能被预先给予的VR1受体阻断剂capsazepine取消。④辣椒素可剂量依赖性降低AngII诱导的内皮细胞凋亡,减弱AngII增加内皮细胞caspase-3活性,此作用能被预先给予的capsazepine和CGRP8-37取消。⑤辣椒素可以上调抗凋亡分子Bcl-2 mRNA的表达,下调促凋亡分子Bax mRNA的表达,此作用能被预先给予的capsazepine和CGRP8-37取消。结论 AngⅡ能抑制内皮细胞合成与释放CGRP;用辣椒素促进内皮源性CGRP的表达能对抗AngⅡ诱导的内皮细胞凋亡。
[Abstract]:Objective to investigate the antiapoptotic effect of endothelium-derived calcitonin gene-related peptide on angiotensin II induced endothelial cells. Methods HUVEC-12 was incubated with different doses of AngII to investigate the dose-effect and time-dependent relationship, and the possible mechanism of Losartan, a specific AT1 receptor blocker, was studied. Real time-PCR was used to detect the level of CGRP mRNA in endothelial cells. The content of CGRP in endothelial cells was detected by radioimmunoassay. Apoptosis was induced by AngII in cultured HUVEC-12, apoptosis was detected by CGRP and capsaicin. Annexin V-FITC flow cytometry was used to detect apoptosis, and caspase-3 activity was detected by RT-PCR to detect the expression of Bcl-2 and Bax mRNA in endothelial cells. Results the expression of 伪-and 尾 -CGRP mRNA and the content of CGRP were decreased in a dose-and time-dependent manner after 1AngII incubation, which could be blocked by losartan in a dose-dependent manner, and the apoptosis of endothelial cells induced by AngII was decreased in a dose-dependent manner. Attenuated AngII increased caspase-3 activity in endothelial cells, which could be inhibited by CGRP receptor blocker CGRP8-37. 3 capsaicin could up-regulate the expression of endothelium-derived 伪-and 尾 -CGRP mRNA in a dose-dependent manner. This effect could be eliminated by pre-administered VR1 receptor antagonist capsazepine. Capsaicin could decrease the apoptosis of endothelial cells induced by AngII in a dose-dependent manner, and decrease the increase of caspase-3 activity of endothelial cells induced by AngII. This effect can be cancelled by pre-administration of capsazepine and CGRP8-37. 5 capsaicin can up-regulate the expression of anti-apoptotic molecule Bcl-2 mRNA and down-regulate the expression of pro-apoptotic molecule Bax mRNA. This effect can be cancelled by pre-administered capsazepine and CGRP8-37. Conclusion Ang 鈪，

本文编号：1941000