This study aimed to elucidate the molecular mechanisms by which berberine protects against cerebral ischemia/reperfusion(I/R)injury.The oxygen-glucose deprivation/reperfusion(OGD/R)PC12 model was established.Cell coun...This study aimed to elucidate the molecular mechanisms by which berberine protects against cerebral ischemia/reperfusion(I/R)injury.The oxygen-glucose deprivation/reperfusion(OGD/R)PC12 model was established.Cell counting kit-8(CCK-8)was used to detect the toxicity of berberine and the viability of PC12 cells.Hoechst 33258 staining and flow cytometry were used to observe the nuclear morphology,and changes of apoptosis and reactive oxygen species(ROS),respectively.Western blotting and immunofluorescence assay were employed to detect autophagy-related proteins[microtubule-associated protein 1A/1B-light chain 3(LC3),P62/SQSTM-1,Beclin-1]and endoplasmic reticulum(ER)stress-related markers[glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),Bcl-2-associated X(Bax)and cleaved caspase-3].The GFP-RFP-LC3 adenovirus was used to assay the change of autophagic flux.Our results showed that berberine could increase the viability of PC12 cells,decrease the concentrations of ROS after OGD/R treatment,and suppress OGD/R-induced ER stress and autophagy.Moreover,the results revealed the involvement of the mammalian target of rapamycin(mTOR)pathway in the induction of autophagy,and berberine could activate the phosphorylation of mTOR and thus mitigate autophagy.In conclusion,our study suggested that berberine may protect against OGD/R-induced apoptosis by regulating ER stress and autophagy,and it holds promises in the treatment of cerebral I/R injury.展开更多
基金This work was supported by grants from the National Natural Science Foundation of China(No.81360199)Science and Technology Department of Guizhou Province(Guizhou Specific Grant[2019]4008)+2 种基金Science and Technology Plan Project of Guizhou Province(Basic Science and Technology Cooperation[2020]1Z060)Science and Technology Fund Project of Guizhou Health and Health Commission(No.gzwjkj2019-1-039)Science and Technology Fund Project of Southwest Guizhou Autonomous Prefecture(2019-1-10).
文摘This study aimed to elucidate the molecular mechanisms by which berberine protects against cerebral ischemia/reperfusion(I/R)injury.The oxygen-glucose deprivation/reperfusion(OGD/R)PC12 model was established.Cell counting kit-8(CCK-8)was used to detect the toxicity of berberine and the viability of PC12 cells.Hoechst 33258 staining and flow cytometry were used to observe the nuclear morphology,and changes of apoptosis and reactive oxygen species(ROS),respectively.Western blotting and immunofluorescence assay were employed to detect autophagy-related proteins[microtubule-associated protein 1A/1B-light chain 3(LC3),P62/SQSTM-1,Beclin-1]and endoplasmic reticulum(ER)stress-related markers[glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),Bcl-2-associated X(Bax)and cleaved caspase-3].The GFP-RFP-LC3 adenovirus was used to assay the change of autophagic flux.Our results showed that berberine could increase the viability of PC12 cells,decrease the concentrations of ROS after OGD/R treatment,and suppress OGD/R-induced ER stress and autophagy.Moreover,the results revealed the involvement of the mammalian target of rapamycin(mTOR)pathway in the induction of autophagy,and berberine could activate the phosphorylation of mTOR and thus mitigate autophagy.In conclusion,our study suggested that berberine may protect against OGD/R-induced apoptosis by regulating ER stress and autophagy,and it holds promises in the treatment of cerebral I/R injury.
