Downregulation of Connexin 32 Attenuates Hypoxia/Reoxygenation Injury in Liver Cells
Downregulation of Connexin 32 Attenuates Hypoxia/Reoxygenation Injury in Liver Cells
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DOI：
10.1002/jbt.21684
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发表时间：
2015-04-01
影响因子：
3.6
通讯作者：
Li, Shangrong
中科院分区：
文献类型：
--
作者：
Wang, Ren;Huang, Fei;Li, Shangrong
细胞间隙连接通讯参与器官缺血再灌注损伤。连接蛋白是对间隙连接的功能至关重要的蛋白质。为了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。BRL-3A大鼠肝细胞，内源性表达连接蛋白Cx 32和Cx43，被用来模拟肝脏IR损伤的过程。间隙连接活性的抑制是通过遗传学方法实现的，使用Cx 32特异性小干扰RNA（siRNA），或通过化学方法实现的，使用药理学抑制剂、油酰胺和18-GA。BRL-3A细胞进行H/R表现出减少的细胞存活和病理指示IR损伤。Cx 32特异性siRNA、油酰胺和18-GA分别降低间隙连接通透性，如降落伞试验所评估。用Cx 32特异性siRNA预处理增加细胞存活。用油酸酰胺或18-GA预处理不能提高细胞存活率。通过Cx 32基因沉默调节间隙连接保护BRL-3A肝细胞免受H/R。
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--GA. BRL-3A cells subjected to H/R exhibited reduced cell survival and pathologies indicative of IR injury. Cx32-specific siRNA, oleamide, and 18--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--GA did not improve cell survival. Modulating gap junction by Cx32 gene silencing protected BRL-3A liver cells from H/R.