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[Effect of ozonized saline on signaling passway of Keap1-Nrf2-ARE in rat hepatocytes]
Dan-dan Qu1, Fu-jiang Peng, Li Liu
1Department of Infectious Diseases, Nanfang Hospital of Southern Medical University, Guangzhou 510515, China.
Objective:
To study the effect of ozonized saline on the activation of the Keap1-Nrf2-ARE signaling pathway in rat liver cells.
Methods:
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.
Results:
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.
Conclusion:
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.
Insights
Ozonized saline preconditioning reduces carbon tetrachloride-induced liver injury in rats by activating the Keap1-Nrf2-ARE pathway. This treatment enhances antioxidant defenses and strengthens Nrf2 expression, offering a protective effect against oxidative damage.
Area of Science:
- Biochemistry
- Cell Biology
- Toxicology
Background:
- Carbon tetrachloride (CCl4) is a known hepatotoxin that induces significant liver injury through oxidative stress.
- The Keap1-Nrf2-ARE signaling pathway plays a crucial role in cellular defense against oxidative damage.
- Ozonated compounds have demonstrated therapeutic potential in various medical applications.
Purpose of the Study:
- To investigate the protective effects of ozonized saline (OS) on CCl4-induced liver injury in a rat model.
- To determine if OS activates the Keap1-Nrf2-ARE signaling pathway.
- To evaluate the impact of OS on antioxidant capacity and liver enzyme levels.
Main Methods:
- Male Sprague-Dawley rats were divided into four groups: ozonized saline (OS), model (CCl4), ozonized saline control (OSC), and normal control (NC).
- OS group received daily intravenous OS for 15 days followed by CCl4 injection; model group received oxygen saline followed by CCl4.
- Liver injury markers (ALT, AST), antioxidant parameters (TAOC, GSH, CAT, GPx), and Nrf2 expression (Western Blot, immunofluorescence) were assessed.
Main Results:
- Ozonized saline significantly reduced serum ALT and AST levels in the OS group compared to the model group (P < 0.01).
- OS treatment led to significantly higher levels of TAOC, CAT, GPx, and GSH in liver tissues compared to the model group (P < 0.01 or P < 0.05).
- Ozonized saline administration was shown to enhance Nrf2 expression in liver cells.
Conclusions:
- Preconditioning with ozonized saline effectively mitigates CCl4-induced liver injury in rats.
- Ozonized saline acts as a novel activator of the Keap1-Nrf2-ARE signaling pathway.
- Activation of this pathway by ozonized saline reduces oxidative damage from reactive oxygen species (ROS).
