Wang Ren, Huang Fei, Chen Zhangyan, Li Shangrong
Department of Anaesthesiology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People's Republic of China.
J Biochem Mol Toxicol. 2015 Apr;29(4):189-97. doi: 10.1002/jbt.21684. Epub 2014 Dec 22.
Gap junction intercellular communication is involved in ischemia-reperfusion (IR) injury of organs. Connexins are proteins that are critical to the function of gap junctions. To clarify the role of gap junctions in IR injury in liver cells, the function of gap junctions was modulated in an in vitro hypoxia/reoxygenation (H/R) model. BRL-3A rat liver cells, endogenously expressing connexins Cx32 and Cx43, were used to model the process of hepatic IR injury. Suppression of gap junction activity was achieved genetically, using Cx32-specific small interfering RNA (siRNA), or chemically, with pharmacological inhibitors, oleamide, and 18-α-GA. BRL-3A cells subjected to H/R exhibited reduced cell survival and pathologies indicative of IR injury. Cx32-specific siRNA, oleamide, and 18-α-GA, respectively, decreased gap junction permeability, as assessed by the parachute assay. Pretreatment with Cx32-specific siRNA increased cell survival. Pretreatment with oleamide or 18-α-GA did not improve cell survival. Modulating gap junction by Cx32 gene silencing protected BRL-3A liver cells from H/R.
间隙连接细胞间通讯参与器官的缺血再灌注（IR）损伤。连接蛋白是对间隙连接功能至关重要的蛋白质。为阐明间隙连接在肝细胞IR损伤中的作用，在体外缺氧/复氧（H/R）模型中调节间隙连接的功能。使用内源性表达连接蛋白Cx32和Cx43的BRL-3A大鼠肝细胞来模拟肝脏IR损伤过程。通过使用Cx32特异性小干扰RNA（siRNA）进行基因抑制，或使用药理抑制剂油酰胺和18-α-甘草次酸进行化学抑制，来实现间隙连接活性的抑制。经历H/R的BRL-3A细胞表现出细胞存活率降低以及指示IR损伤的病理变化。通过降落伞试验评估，Cx32特异性siRNA、油酰胺和18-α-甘草次酸分别降低了间隙连接通透性。用Cx32特异性siRNA预处理可提高细胞存活率。用油酰胺或18-α-甘草次酸预处理并不能改善细胞存活率。通过Cx32基因沉默调节间隙连接可保护BRL-3A肝细胞免受H/R损伤。
