环孢素 A-对微血管高通透性的保护作用不依赖于钙调神经磷酸酶。

Cyclosporine A--protection against microvascular hyperpermeability is calcineurin independent.

机构信息

Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 2401 South 31st St., Temple, TX 76508, USA.

出版信息

Am J Surg. 2010 Apr;199(4):542-8. doi: 10.1016/j.amjsurg.2009.11.002.

DOI:10.1016/j.amjsurg.2009.11.002

PMID:20359571

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2864542/

Abstract

BACKGROUND

Mitochondria-mediated apoptotic signaling contributes to microvascular hyperpermeability. We hypothesized that cyclosporine A (CsA), which protects mitochondrial transition pores, would attenuate hyperpermeability independent of its calcineurin inhibitory property.

METHODS

Hyperpermeability was induced in microvascular endothelial cell monolayers using proapoptotic BAK or active caspase-3 after CsA or a specific calcineurin inhibitor, calcineurin autoinhibitory peptide (CIP), treatment. Permeability was measured based on fluorescein isothiocyanate-albumin flux across the monolayers. Mitochondrial transmembrane potential (MTP) was determined using 5,5',6,6'-tetrachoro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide. Mitochondrial release of cytochrome c was measured using an enzyme-linked immunosorbent assay and caspase-3 activity fluorometrically.

RESULTS

CsA-attenuated (10 nmol/L) but not CIP-attenuated (100 mumol/L) BAK induced hyperpermeability (P < .05), CsA- but not CIP-attenuated BAK induced a decrease in MTP and an increase in cytochrome c levels and caspase-3 activity (P < .05). CsA and CIP were ineffective against caspase-3-induced hyperpermeability.

CONCLUSIONS

CsA attenuated hyperpermeability by protecting MTP, thus preventing mitochondria-mediated apoptotic signaling. The protective effect of CsA is independent of calcineurin inhibition.

摘要

背景

线粒体介导的凋亡信号通路参与了微血管通透性的增加。我们假设环孢素 A（CsA），作为一种可以保护线粒体通透转换孔的物质，能够通过不依赖于其钙调神经磷酸酶抑制特性的方式减轻通透性的增加。

方法

使用促凋亡的 BAK 或活性半胱天冬酶-3 诱导微血管内皮细胞单层的通透性增加，之后用 CsA 或特定的钙调神经磷酸酶抑制剂钙调神经磷酸酶自身抑制肽（CIP）进行处理。通过测定单层细胞中荧光素异硫氰酸酯-白蛋白的通量来测量通透性。使用 5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑基碳酰二亚胺碘化物来测定线粒体跨膜电位（MTP）。通过酶联免疫吸附试验和荧光法测定线粒体释放细胞色素 c 和半胱天冬酶-3 的活性。

结果

CsA 减弱（10 nmol/L）但 CIP 没有减弱（100 mumol/L）BAK 诱导的通透性增加（P <.05），CsA 减弱但 CIP 没有减弱 BAK 诱导的 MTP 下降以及细胞色素 c 水平和半胱天冬酶-3 活性的增加（P <.05）。CsA 和 CIP 对 caspase-3 诱导的通透性增加没有作用。