Butein Alleviates Cerebral Ischemic Reperfusion Injury by Modulating the TLR4/NF-κB Pathway

Abstract

Background: Cerebral ischemia-reperfusion (CI/R) injury poses significant threats to human health due to its high disability, morbidity, and mortality rates. This study investigates the effects of Butein in oxidative stress-induced injuries in PC12 cells and its underlying mechanism on CI/R injury. Methods: An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established using PC12 cells to examine the impact of Butein on CI/R injury. Cell viability and apoptosis were assessed using Cell Counting Kit-8 (CCK-8) and flow cytometry (FCM) techniques, respectively. Protein expressions of toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB), B-cell lymphoma-2 (Bcl-2) and BCL-2-associated X protein(Bax) were determined by Western blot assay. Additionally, Buteins effects on the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS) in OGD/R cells were evaluated using xanthine oxidase, thiobarbituric acid methods, and flow cytometry, respectively. Results: Butein augmented the viability of PC12 cells under OGD/R conditions and diminished cell apoptosis. It notably suppressed ROS and MDA production while increased SOD levels. Furthermore, Butein treatment led to a marked reduction in TLR4 and NF-κB expressions. Notably, combining Butein with si-TLR4 enhanced the protective effect of Butein against CI/R injury. Conclusions: Butein mitigates oxidative stress and reduces apoptosis by inhibiting the TLR4/NF-κB pathway, offering potential therapeutic benefits against CI/R injury.