Cryptotanshinone alleviates hypoxia/reoxygenation-induced neuronal damage by regulating the Hippo/YAP signaling pathway

CHEN Liang; MAN Xiaorui; BIAN Junli

doi:10.3969/j.issn.1002-7386.2025.01.008

Abstract：Objective To investigate the impacts of cryptotanshinone(CPT)on hypoxia/reoxygenation(H/R)-induced neuronal damage by regulating the Hippo/Yes-associated protein(YAP)signaling pathway.Methods Human cortical neurons(HCN)were treated with blank control,or induced with H/R,H/R+low-dose CPT(25 μg/mL CPT),H/R+medium-dose CPT(50 μg/mL CPT),H/R+high-dose CPT(80 μg/mL CPT),H/R+YAP inhibitor Verteporfin(VTPF,5 µmol/L)and H/R+high-dose CPT+VTPF(80 μg/mL CPT+5 µmol/L VTPF).Cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.Relative levels of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA)were measured using commercial kits.The 2',7'-dichlorofluorescein diacetate(DCFH-DA)probe was applied to detect the level of reactive oxygen species(ROS)in HCN cells.Western blot was applied to detect the protein expressions of proteins in the Hippo/YAP signaling pathway and those associated with apoptosis.Results Compared with those treated with blank control,HCN cells induced with H/R presented significantly lower OD450,SOD level and protein expression of p-YAP(P＜0.05),and higher LDH,MDA and ROS levels,apoptotic rate and protein expression of YAP(P＜0.05).Compared with those induced with H/R,HCN cells induced with H/R and low-dose,medium-dose and high-dose CPT presented significantly higher OD450,SOD level and protein expression of p-YAP(P＜0.05),and lower LDH,MDA and ROS levels,apoptotic rate and protein expression of YAP in a dose-dependent manner(P＜0.05).Opposite trends were observed in HCN cells induced with H/R plus VTPF(P＜0.05).VTPF treatment reversed the protective effect of high-dose CPT on H/R-induced neuronal damage.Conclusion CPT improves neuronal damage induced by hypoxia/reoxygenation(H/R)by activating the Hippo/YAP signaling pathway.