Silencing TRPV4 partially reverses the neurotoxic effects caused by excess Ketamine

抄録

Excessive use of Ketamine (KET) has a neurotoxic effect on the brain. This study explored the effect of Transient Receptor Potential Vanilloid 4 (TRPV4) on KET-induced neurotoxicity in the hippocampus. We extracted and identified rat hippocampal neuronal cells. The hippocampal neurons were treated with different concentrations (0, 0.1, 1, 10, 100, 300 and 1000 μmol/L) of KET (6, 12 and 24 hr). Cell viability was detected by cell counting Kit-8 (CCK-8), and TRPV4 expression was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and western blot. After silencing TRPV4, we tested cell viability and apoptosis. The contents of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), and catalase (CAT) were detected by colorimetry, and the contents of TNF-α, IL-1β, IL-6 and reactive oxygen species (ROS) were detected by Enzyme-Linked ImmunoSorbent Assay (ELISA). Finally, the expression levels of apoptosis-related proteins Bcl-2, Bax and Cleaved caspase-3, and phosphorylated-p65 (p-65), p65, phosphorylated-IκBα (p-IκBα) and IκBα were detected by qRT-PCR and western blot. KET inhibited the viability of hippocampal neurons in a dose-dependent manner, and up-regulated TRPV4 expression. SiTRPV4 inhibits KET-induced decrease in cell viability and promotes apoptosis. SiTRPV4 reduced MDA and ROS content, increased SOD, GSH and CAT levels. The release of proinflammatory factors TNF-α, IL-1β and IL-6 was also inhibited by siTRPV4. In addition, siTRPV4 up-regulated KET-induced Bcl-2 expression in hippocampal neurons, down-regulated Bax and Cleaved caspase-3, and inhibited the activation of the inflammatory pathway. Silencing TRPV4 partially reverses the neurotoxic effects induced by KET through regulating apoptosis-related proteins and p65/IκBα pathway.