Dexmedetomidine Regulates Autophagy Via AMPK/mTOR Pathway To Improve SH-SY5Y/APP695 Cells Damage Induced By High Glucose

Objective To observe the protective effect of dexmedetomidine(DEX)on the SH-SY5Y/APP695(SAS)cells injury induced by high glucose,and explore its possible mechanism.Methods1.SAS cells were cultured at different glucose concentrations and incubation time to establish the model of SAS cells injury induced by high glucose.The cell viability was detected by CCK-8,and the appropriate glucose concentration and treatment time were determined to establish high-glucose-induced cell injury model.2.The model of SAS cells injury induced by high glucose was treated with different concentrations of DEX to explore the most protective concentration of DEX against high glucose injury.3.In order to clarify the protective effect of DEX against high-glucose-induced injury,SAS cells were subjected to control group(Control),DEX group(DEX),high glucose group(HG)and high glucose + DEX group(HG+DEX).The cell viability was assessed by CCK-8.The morphological changes of cells were observed under microscope,and the apoptosis rate was analyzed by TUNEL fluorescence staining and flow cytometry.4.SAS cells were divided into control group(Control),high glucose group(HG),high glucose+DEX group(HG+DEX),high glucose+rapamycin(RAPA)group(HG+RAPA),high glucose+DEX+RAPA group(HG+DEX+RAPA),high glucose+3-MA group(HG+3-MA)and high glucose+DEX+3-MA group(HG+DEX+3-MA).Cell damage was detected by cell viability which explored through CCK-8 and cell morphological changes which observed under microscope.Apoptosis rate was analyzed by TUNEL fluorescence staining and flow cytometry.Autophagy activity was evaluated by MDC staining.And the expression of autophagy protein p62,Beclin-1 and LC3II/LC3 I was detected by Western blot.5.SAS cells were divided into control group(Control),DEX group(DEX),high glucose group(HG)and high glucose+DEX group(HG+DEX)to study the mechanism of DEX on autophagy of SAS cells in high glucose environment.The relative expressions of p-AMPK,AMPK,p-m TOR and m TOR were detected by Western blot.6.SAS cells were divided into control group(Control),high glucose group(HG),high glucose+DEX group(HG+DEX)and high glucose+DEX+compound C group(HG+DEX+CC)to explore how DEX regulate the level of autophagy on high-glucose-injured cells.Cell damage was detected by the cell viability which explored through CCK-8 and cell morphological changes which observed under microscope,.Apoptosis rate was evaluated by TUNEL fluorescence staining,flow cytometry and the expression level of apoptosis proteins Caspase 3,BAX and Bcl-2.The level of autophagy was detected by the expression of autophagy protein p62,Beclin-1 and LC3II/LC3 I and MDC stainingResult1.DEX attenuated high-glucose-induced injury of SAS cells.1)The damage of SAS cells induced by high glucose was time-and dose-dependent.2)DEX attenuated high-glucose-induced injury of SAS cells and improve.cell morphology and viability.2.DEX attenuated high-glucose-induced injury of SAS cells through upregulation of autophagy.1)High glucose caused the decline of the autophagy activity in SAS cells.2)DEX upregulated the level of autophagy in SAS cells injured by high glucose.3)The autophagy activity affected the cell activity in model of SAS cells injury induced by high glucose.3.DEX regulated autophagy of SAS cells in high glucose environment through AMPK/m TOR pathway.1)High glucose inhibited AMPK/m TOR pathway in SAS cells.2)DEX activated AMPK/m TOR pathway in SAS cells incubated with high glucose and increased the level of autophagy.ConclusionDEX could protect SAS cells against the high-glucose-induced damage by upregulating the level of autophagy through AMPK/m TOR pathway.