Tian K Y, Liu X J, Xu J D, Deng L J, Wang G
Department of Anesthesiology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
Department of Anesthesia, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Braz J Med Biol Res. 2015 May;48(5):401-7. doi: 10.1590/1414-431X20144107. Epub 2015 Mar 6.
Recent studies have revealed that an intrinsic apoptotic signaling cascade is involved in vascular hyperpermeability and endothelial barrier dysfunction. Propofol (2,6-diisopropylphenol) has also been reported to inhibit apoptotic signaling by regulating mitochondrial permeability transition pore (mPTP) opening and caspase-3 activation. Here, we investigated whether propofol could alleviate burn serum-induced endothelial hyperpermeability through the inhibition of the intrinsic apoptotic signaling cascade. Rat lung microvascular endothelial cells (RLMVECs) were pretreated with propofol at various concentrations, followed by stimulation with burn serum, obtained from burn-injury rats. Monolayer permeability was determined by transendothelial electrical resistance. Mitochondrial release of cytochrome C was measured by ELISA. Bax and Bcl-2 expression and mitochondrial release of second mitochondrial-derived activator of caspases (smac) were detected by Western blotting. Caspase-3 activity was assessed by fluorometric assay; mitochondrial membrane potential (Δψm) was determined with JC-1 (a potential-sensitive fluorescent dye). Intracellular ATP content was assayed using a commercial kit, and reactive oxygen species (ROS) were measured by dichlorodihydrofluorescein diacetate (DCFH-DA). Burn serum significantly increased monolayer permeability (P<0.05), and this effect could be inhibited by propofol (P<0.05). Compared with a sham treatment group, intrinsic apoptotic signaling activation - indicated by Bax overexpression, Bcl-2 downregulation, Δψm reduction, decreased intracellular ATP level, increased cytosolic cytochrome C and smac, and caspase-3 activation - was observed in the vehicle group. Propofol not only attenuated these alterations (P<0.05 for all), but also significantly decreased burn-induced ROS production (P<0.05). Propofol attenuated burn-induced RLMVEC monolayer hyperpermeability by regulating the intrinsic apoptotic signaling pathway.
最近的研究表明，一种内在的凋亡信号级联反应与血管通透性增加和内皮屏障功能障碍有关。据报道，丙泊酚（2,6 - 二异丙基苯酚）可通过调节线粒体通透性转换孔（mPTP）开放和半胱天冬酶 - 3激活来抑制凋亡信号。在此，我们研究了丙泊酚是否能通过抑制内在凋亡信号级联反应来减轻烧伤血清诱导的内皮细胞高通透性。用不同浓度的丙泊酚预处理大鼠肺微血管内皮细胞（RLMVECs），然后用从烧伤大鼠获得的烧伤血清进行刺激。通过跨内皮电阻测定单层通透性。用ELISA法测定细胞色素C的线粒体释放。通过蛋白质印迹法检测Bax和Bcl - 2表达以及第二线粒体衍生的半胱天冬酶激活剂（smac）的线粒体释放。用荧光测定法评估半胱天冬酶 - 3活性；用JC - 1（一种电位敏感荧光染料）测定线粒体膜电位（Δψm）。使用商业试剂盒测定细胞内ATP含量，并用二氯二氢荧光素二乙酸酯（DCFH - DA）测量活性氧（ROS）。烧伤血清显著增加单层通透性（P<0.05），而丙泊酚可抑制这种作用（P<0.05）。与假处理组相比，在溶剂对照组中观察到内在凋亡信号激活，表现为Bax过表达、Bcl - 2下调、Δψm降低、细胞内ATP水平降低、细胞色素C和smac胞质增加以及半胱天冬酶 - 3激活。丙泊酚不仅减轻了这些改变（所有P<0.05），而且显著降低了烧伤诱导的ROS产生（P<0.05）。丙泊酚通过调节内在凋亡信号通路减轻烧伤诱导的RLMVEC单层高通透性。
