熊果苷调控Notch1/Hes1通路对缺氧/复氧诱导的心肌细胞损伤影响的研究OA

Objective To investigation the effect of arbutin on hypoxia/reoxygenation(H/R)-induced injury of cardiomyocytes(H9C2)through the neurogenic locus notch homolog protein 1(Notch 1)/hairy and enhancer of split 1(Hes1)signaling pathway.Methods An H/R-induced H9C2 cell injury model was established.The optimal intervention concentration of arbutin was screened by pretreating cells with various concentrations(12.50-200.00μmol·L-1).H9C2 cells were divided into a normal group,a model group,experimental-L,M,H groups(25.00,50.00,and 100.00 μmol·L-1 arbutin,respectively),and an inhibitor group { 100.00 μmol·L-1 arbutin+10.00μmol·L-1N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester(DAPT)}.Cells in the normal group were cultured under normal conditions,while H/R models were established in the remaining groups.All treatment groups received arbutin pretreatment at concentrations ranging from 12.50 to 200.00 μmol·L-1 for 24 hours before modeling.Cell proliferation was detected by the CCK-8 assay,and apoptosis was assessed by TUNEL assay;mitochondrial membrane potential was measured by JC-1;reactive oxygen species(ROS),lactate dehydrogenase(LDH),glutathione peroxidase(GSH-Px),malondialdehyde(MDA),and superoxide dismutase(SOD)levels were detected using kits.Expression of Notch1,Hes1,Bcl-2,Bax,and caspase-3 was detected by Western blot.Results Arbutin at concentrations of 25.00-200.00 μmol·L-1 significantly increased the survival rate of H/R-induced H9C2 cells(P＜0.05).Arbutin at 25.00,50.00,and 100.00 μmol·L-1 was selected as the low,medium,and high doses for subsequent experiments.The creatine kinase isoenzyme MB(CK-MB)levels in the normal group,model group,experimental-H group,and inhibitor group were(2.45±0.39),(8.72±0.82),(3.56±0.42),and(5.48±0.57)U·mL-1,respectively;the cTnI levels were(22.58±2.23),(68.34±5.96),(30.12±3.26),and(49.62±5.15)ng·L-1,respectively;the LDH levels were(53.42±5.44),(163.72±13.41),(65.45±6.57),and(98.33±9.85)U·L-1,respectively;the relative ROS levels were 1.01±0.12,3.34±0.35,1.46±0.18,and 2.24±0.23,respectively;the SOD levels were(96.23±8.12),(24.31±2.24),(78.39±7.93),and(43.78±4.49)U·mg-1,respectively;the GSH-Px levels were(74.82±7.55),(16.42±1.83),(61.32±6.29),and(35.72±3.38)U·mg-1,respectively;the MDA levels were(15.45±2.03),(58.33±5.57),(21.56±2.24),and(42.47±4.13)μmol·L-1,respectively;the apoptosis rates were(2.45±0.39)％,(29.86±3.25)％,(9.86±0.91)％,and(21.34±2.25)％,respectively;the red/green fluorescence intensity ratios were(5.78±0.65)％,(1.42±0.20)％,(4.89±0.54)％,and(2.85±0.33)％,respectively;the Notch1 protein expression levels were 1.27±0.12,0.34±0.03,1.05±0.08,and 0.54±0.05,respectively;the Hes1 protein expression levels were 0.95±0.10,0.25±0.03,0.81±0.08,and 0.57±0.05,respectively;the Bcl-2 protein expression levels were 1.01±0.11,0.31±0.03,0.89±0.08,and 0.61±0.06,respectively;the Bax protein expression levels were 0.45±0.04,1.34±0.12,0.61±0.06,and 0.93±0.09,respectively;and the caspase-3 protein expression levels were 0.23±0.03,0.91±0.09,0.37±0.04,and 0.64±0.06,respectively.Comparisons between the normal group and the model group,between the model group and the experimental-L,M,H groups,and between the experimental-H group and the inhibitor group all showed statistically significant differences for the above indicators(all P＜0.05).Conclusion Arbutin alleviates H/R-induced H9C2 cell injury by activating the Notch1/Hes1 pathway and inhibiting oxidative stress.