Volatile anesthetics induce caspase-dependent, mitochondria-mediated apoptosis in human T lymphocytes in vitro
Volatile anesthetics induce caspase-dependent, mitochondria-mediated apoptosis in human T lymphocytes in vitro
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DOI：
10.1097/00000542-200506000-00014
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发表时间：
2005-06-01
期刊：
影响因子：
8.8
通讯作者：
Pannen, BHJ
中科院分区：
文献类型：
--
作者：
Loop, T;Dovi-Akue, D;Pannen, BHJ
背景：挥发性麻醉剂在手术中调节淋巴细胞功能，这损害了术后免疫能力。目前的工作是研究挥发性麻醉剂是否会诱导人T淋巴细胞凋亡以及可能使用的凋亡信号通路。方法：研究七氟醚、异氟醚和地氟醚对体外培养的人CD3+ T淋巴细胞和Jurkat T细胞的影响。绿色荧光蛋白-膜联蛋白V和DiOC(6)-荧光染色后，采用How细胞术检测细胞凋亡和线粒体膜电位。通过切割荧光效应caspase底物Ac-DEVD-AMC和抗caspase-3 Western blotting检测caspase 3的活性和蛋白水解过程。用抗细胞色素c Western blotting和酶联免疫吸附法研究细胞分离后线粒体细胞色素c的释放。结果：七氟醚和异氟醚诱导人T淋巴细胞凋亡呈剂量依赖性。相反地氟醚对细胞凋亡无促进作用。七氟醚使用的凋亡信号通路涉及线粒体膜电位的破坏和细胞色素c从线粒体释放到细胞质中。此外，作者观察到无活性的p32 procaspase 3被裂解为有活性的p17片段，caspase-3样活性增加，caspase-3底物聚adpriose -聚合酶被裂解。七氟醚诱导的细胞凋亡被通用caspase抑制剂Z-VAD.fmk阻断。死亡信号不是通过Fas/CD95受体途径介导的，因为无论是抗Fas/CD95受体拮抗剂还是FADD缺陷或caspase-8缺陷都不能减弱七氟醚介导的细胞凋亡。结论：七氟醚和异氟醚通过增加线粒体膜通透性和caspase-3活化诱导T淋巴细胞凋亡，但不依赖于死亡受体信号。
Background: Volatile anesthetics modulate lymphocyte function during surgery, and this compromises postoperative immune competence. The current work was undertaken to examine whether volatile anesthetics induce apoptosis in human T lymphocytes and what apoptotic signaling pathway might be used.Methods: Effects of sevoflurane, isoflurane, and desflurane were studied in primary human CD3+ T lymphocytes and Jurkat T cells in vitro. Apoptosis and mitochondrial membrane potential were assessed using How cytometry after green fluorescent protein-annexin V and DiOC(6)-fluorochrome staining. Activity and proteolytic processing of caspase 3 was measured by cleaving of the fluorogenic effector caspase substrate Ac-DEVD-AMC and by anti-caspase-3 Western blotting. Release of mitochondrial cytochrome c was studied after cell fractionation using anti-cytochrome c Western blotting and enzyme-linked immunosorbent assays.Results: Sevoflurane and isoflurane induced apoptosis in human T lymphocytes in a dose-dependent manner. By contrast, desflurane did not exert any proapoptotic effects. The apoptotic signaling pathway used by sevoflurane involved disruption of the mitochondrial membrane potential and release of cytochrome c from mitochondria to the cytosol. In addition, the authors observed a proteolytic cleavage of the inactive p32 procaspase 3 to the active p17 fragment, increased caspase-3-like activity, and cleavage of the caspase-3 substrate poly-ADPribose-polymerase. Sevoflurane-induced apoptosis was blocked by the general caspase inhibitor Z-VAD.fmk. Death signaling was not mediated via the Fas/CD95 receptor pathway because neither anti-Fas/CD95 receptor antagonism nor FADD deficiency or caspase-8 deficiency were able to attenuate sevoflurane-mediated apoptosis.Conclusion: Sevoflurane and isoflurane induce apoptosis in T lymphocytes via increased mitochondrial membrane permeability and caspase-3 activation, but independently of death receptor signaling.