Abstract
Objective To verify whether ginsenoside Rg1 alleviates liver hepatic ischemia-reperfusion injury (IRI) in mice by upregulating the expression of Yes-associated protein (YAP) through estrogen receptor alpha pathway. Methods The whole hepatic IRI model and the local (70%) hepatic IRI model were established, respectively. The whole hepatic IRI model was used to observe the survival curve of mice, and the mouse models with 70% hepatic IRI were used to explore the mechanism of liver injury about Rg1 in hepatic IRI. Wild-type C57BL/6 mice were randomly divided into some groups: (1) the whole hepatic IRI model group: the survival rate of mice was observed at 0, 30, 60, 90, and 120 min after ischemia and Rg1 intervention (90 min after ischemia), with 10 mice in each group, and (2) the 70% hepatic IRI model group: sham operation group, I/R model group, verteporfin (VP) group, doxycycline (Doxy) group, 17β-estradiol (E2) group, clomiphene (Clom) group, and Rg1 group with 6 mice in each group. The level of serum alanine aminotransferase (ALT) was measured by enzyme labeling instrument, the degree of liver injury was analyzed after hematoxylin-eosin (HE) staining, and the function of mitochondria was detected in fresh liver tissue, including mitochondrial membrane potential with JC-1 (5,5′,6,6′-tetrachloro1,1′,3,3′-tetramethylbenzimidazolylcarbocyanine iodide), adenosine triphosphate (ATP), and mitochondrial reactive oxygen species (ROS), and the expression of YAP and estrogen receptor alpha (ERα) genes and proteins were detected by real‐time reverse‐transcriptase polymerase chain reaction (RT-PCR) and Western blot. Results The whole hepatic IRI model showed that the survival rate of mice decreased with the prolongation of ischemia time. IRI model mice showed mitochondrial damage, JC-1 red/green fluorescence value and ATP significantly decreased, and ROS production increased; in comparison, in the Doxy and E2 intervention group, JC-1 red/green fluorescence value and ATP production increased and ROS downregulated, indicating that mitochondrial function returned to normal. The level of serum ALT showed that the liver enzyme increased with the time of reperfusion and decreased gradually after 6 hours. The results of Western blot and PCR showed that the expression of YAP and ERα showed the same trend. The IRI model mice were observed after 90 minutes of ischemia and 6 hours of reperfusion. Compared with the corresponding sham group, the expression of YAP in the liver tissue of the Doxy group, E2 group, and Rg1 intervention group increased, and the expression of ERα in the E2 group and Rg1 group increased. HE staining showed that a large number of inflammatory cell infiltration could be seen in the liver tissue of the model group, but it decreased in the Doxy and E2 intervention groups. Conclusion Ginsenoside Rg1 exerts an estrogenic effect by activating ERα, upregulating the expression of YAP, reducing liver oxidative stress injury, and inhibiting mitochondrial injury to protect the liver from ischemia-reperfusion injury in mice.
Highlights
ischemia-reperfusion injury (IRI) is an exogenous non-antigen-dependent local inflammatory reaction caused by hypoxic stress, which often occurs in orthotopic liver transplantation (OLT), vascular embolism, hemorrhagic shock, acute ischemic trauma, druginduced liver injury, and other diseases. e repair of liver injury mediated by IR is the difficulty in the treatment of these diseases
We established a mouse model of hepatic IRI. rough drug preconditioning, we investigated the expression of Yes-associated protein (YAP) and the protective effects of estrogen on liver injury and liver fibrosis and studied the mechanism of Rg1 alleviating hepatic IRI by upregulating the expression of YAP by ERα
When ischemia and hypoxia occur in the liver, the transport of oxygen and nutrients is impaired, and the lack of oxygen as the terminal electron carrier of the mitochondrial respiratory chain will immediately interrupt the mitochondrial electron flow, resulting in respiratory chain dysfunction, increase in the intracellular NADH/NAD+ ratio, termination of oxidative phosphorylation, and the depletion of cellular adenosine triphosphate (ATP). e respiratory chain of hepatocytes is interrupted, and the mitochondria that lose the supply of oxygen and glucose cannot produce enough ATP supply, which will lead to the production of reactive oxygen species (ROS), and the accumulation of toxic substances, accelerated glycolysis, and increased lactic acid formation, resulting in hepatocyte damage [2]
Summary
IRI is an exogenous non-antigen-dependent local inflammatory reaction caused by hypoxic stress, which often occurs in orthotopic liver transplantation (OLT), vascular embolism, hemorrhagic shock, acute ischemic trauma, druginduced liver injury, and other diseases. e repair of liver injury mediated by IR is the difficulty in the treatment of these diseases. IRI is an exogenous non-antigen-dependent local inflammatory reaction caused by hypoxic stress, which often occurs in orthotopic liver transplantation (OLT), vascular embolism, hemorrhagic shock, acute ischemic trauma, druginduced liver injury, and other diseases. E repair of liver injury mediated by IR is the difficulty in the treatment of these diseases. The clinical treatment of hepatic IRI mainly focuses on reducing ischemic injury, controlling. Some studies have confirmed that estrogen has a protective effect on liver injury in IR. Other studies have confirmed that Hippo-YAP pathway plays a key role in liver IR-mediated hepatocyte injury and liver fibrosis, and there is a mutual regulation between estrogen and Hippo-YAP signal pathway. Rough drug preconditioning, we investigated the expression of YAP and the protective effects of estrogen on liver injury and liver fibrosis and studied the mechanism of Rg1 alleviating hepatic IRI by upregulating the expression of YAP by ERα We established a mouse model of hepatic IRI. rough drug preconditioning, we investigated the expression of YAP and the protective effects of estrogen on liver injury and liver fibrosis and studied the mechanism of Rg1 alleviating hepatic IRI by upregulating the expression of YAP by ERα
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