Qu Dan-dan, Peng Fu-jiang, Liu Li, Yang Shu-ling, Guo Ya-bing
Department of Infectious Diseases, Nanfang Hospital of Southern Medical University, Guangzhou 510515, China.
Zhonghua Gan Zang Bing Za Zhi. 2011 May;19(5):367-71. doi: 10.3760/cma.j.issn.1007-3418.2011.05.013.
To study the effect of ozonized saline on the activation of the Keap1-Nrf2-ARE signaling pathway in rat liver cells.
Twenty male Sprague-Dawley rats were randomly divided into ozonized saline (OS) group, model group, ozonized saline control (OSC) group and normal control (NC) group. The rats in OS group and model group were intravenously administered with OS or oxygen saline (5 ml/kg) respectively, once a day for 15 days, and then intraperitoneally injected with CCl4 dissolved in oliver oil. The rats in OSC group were pretreated with OS for 15 days. The rats in NC group were fed normally for 15 days. On the 16th day, the rats in OSC group and NC group were intraperitoneally injected with oliver oil (2 ml/kg) without CCl4. After 24 hours of CCl4 or olive oil intraperitoneal injection, the serum levels of alanine transaminase (ALT) and aspertate aminotransferase (AST) were measured. The liver tissues were also collected for detection of total anti-oxygen capability (TAOC), glutathione (GSH), catalase (CAT), Glutathione peroxidase (GPx). Western Blot was used to detect Nrf2 and immunofluorescence staining assay to display intracelluar distribution of Nrf2.
Compared with the rats in model group,the serum ALT and AST levels of rats in OS group were significantly lower (P < 0.01) ,which were (1240.4 ± 188.2) U/L and (1245.4 ± 176.9) U/L vs (539.8 ± 175.3) U/L and (546.0 ± 130.2) U/L, and the TAOC, CAT, GPx and GSH activity of rats in OS group were significantly higher, which were (0.72 ± 0.24) U/mg, (1.05 ± 0.21) mg/g, (676.9 ± 115.1) U/mg and (45.2 ± 14.3) U/mg vs (1.37 ± 0.19) U/mg, (2.23 ± 0.55) mg/g, (1024.6 ± 162.9) U/mg and (68.2 ± 9.9) U/mg, respectively. In contrast with NC group, pretreatment of OS in OSC group elevated TAOC, CAT, GPx and GSH activity (P < 0.01 or P < 0.05). Ozonized saline can strengthen the Nrf2 expression in liver cells.
Preconditioning injection of ozonized saline can reduce rat's liver injury induced by CCl4. The ozonized saline, as a novel Nrf2 activator, can reduce the oxidative damage of radical oxygen species (ROS) and the deleterious substance by activating the Keap1-Nrf2-ARE signaling pathway and its downstream genes expression.
研究臭氧盐水对大鼠肝细胞中Keap1-Nrf2-ARE信号通路激活的影响。
将20只雄性Sprague-Dawley大鼠随机分为臭氧盐水（OS）组、模型组、臭氧盐水对照（OSC）组和正常对照（NC）组。OS组和模型组大鼠分别静脉注射臭氧盐水或氧盐水（5 ml/kg），每天1次，共15天，然后腹腔注射溶于橄榄油的CCl4。OSC组大鼠用臭氧盐水预处理15天。NC组大鼠正常饲养15天。第16天，OSC组和NC组大鼠腹腔注射不含CCl4的橄榄油（2 ml/kg）。腹腔注射CCl4或橄榄油24小时后，测定血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平。同时收集肝组织检测总抗氧化能力（TAOC）、谷胱甘肽（GSH）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）。采用蛋白质免疫印迹法检测Nrf2，并通过免疫荧光染色法显示Nrf2在细胞内的分布。
与模型组大鼠相比，OS组大鼠血清ALT和AST水平显著降低（P < 0.01），分别为（1240.4 ± 188.2）U/L和（1245.4 ± 176.9）U/L，而模型组为（539.8 ± 175.3）U/L和（546.0 ± 130.2）U/L；OS组大鼠TAOC、CAT、GPx和GSH活性显著升高，分别为（0.72 ± 0.24）U/mg、（1.05 ± 0.21）mg/g、（676.9 ± 115.1）U/mg和（45.2 ± 14.3）U/mg，而模型组为（1.37 ± 0.19）U/mg、（2.23 ± 0.55）mg/g、（1024.6 ± 162.9）U/mg和（68.2 ± 9.9）U/mg。与NC组相比，OSC组用臭氧盐水预处理后TAOC、CAT、GPx和GSH活性升高（P < 0.01或P < 0.05）。臭氧盐水可增强肝细胞中Nrf2的表达。
预先注射臭氧盐水可减轻CCl4诱导的大鼠肝损伤。臭氧盐水作为一种新型的Nrf2激活剂，可通过激活Keap1-Nrf2-ARE信号通路及其下游基因表达，减轻活性氧（ROS）和有害物质的氧化损伤。
