Abstract
Objective:To investigate the effect of Oridonin(Ori)on neuronal damage induced by oxygen glucose deprivation/reoxygenation(OGD/R)by regulating the Hippo/YAP signaling pathway.Methods:OGD/R induced HT22 cells were treated with different concentrations of Ori.The experiment was separated into control group,OGD/R group,low concentration(2 μmol/L)Ori group(Ori-L group),medium concentration(4 μmol/L)Ori group(Ori-M group),high concentration(8 μmol/L)Ori group(Ori-H group)and Ori-H(10 μmol/L)+PY-60(10 μmol/L)group(Ori-H+PY-60 group).The morphological changes of cells was observed under microscope.Edu was applied to detect cell proliferation level.Flow cytometry was applied to detect cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)were applied to measure lactate dehydrogenase(LDH),catalase(CAT)and superoxide dismutase(SOD)levels.Reactive oxygen(ROS)detection kit was applied to measure ROS level.Immunoblotting was applied to determine the expression of B cell lymphoma 2(Bcl-2),Bcl-2 related X protein(Bax),Yes associated protein(YAP)and TAZ proteins.Results:Compared with 0 μmol/L Ori,the cell viability was significantly increased after treatment with 2,4,and 8 μmol/L Ori(P＜0.05),therefore,2,4,and 8 μmol/L Ori was selected for subsequent experiments.Compared with the control group,the OGD/R group showed sparse cell distribution,severe cell damage,shrinkage and swelling of cells,and obvious cell death,and the positive rate of Edu,the levels of CAT and SOD,and the level of Bcl-2 were obviously reduced in OGD/R group(P＜0.05).The apoptosis rate,the levels of LDH and ROS,and the levels of Bax,YAP and TAZ were significantly increased in OGD/R group(P＜0.05).Compared with the OGD/R group,the cells in the Ori-L group,Ori-M group and Ori-H group showed more normal state,intact cell bodies,reduced degree of cell damage and swelling,and reduced number of deaths,and the positive rate of Edu,the levels of CAT and SOD,and the level of Bel-2 were significantly increased in Ori-L group,Ori-M group and Ori-H group(P＜0.05).The apoptosis rate,the levels of LDH and ROS,and the levels of Bax,YAP,and TAZ were obviously reduced in Ori-L group,Ori-M group and Ori-H group(P＜0.05).It was concentration dependent.The YAP activator,PY-60,reversed the protective effect of Ori on OGD/R induced neuronal damage.Conclusion:Ori may improve neuronal damage caused by OGD/R by regulating the Hippo/YAP signaling pathway.关键词
神经元损伤/氧糖剥夺复氧/冬凌草甲素/Hippo/YAP信号通路Key words
neuron damage/oxygen glucose deprivation and reoxygenation/Oridonin/hippo/YAP signaling pathway分类
医药卫生
