摘要
目的探讨利多卡因对氧糖剥夺/复氧(OGD/R)诱导神经细胞损伤的影响及其作用机制。方法常规培养大鼠肾上腺嗜铬细胞(PC12细胞),将培养好的细胞随机分为对照组、模型组、低浓度组、中浓度组、高浓度组、miR-NC+模型组、miR-125b-5p+模型组、anti-miR-NC+高浓度组、anti-miR-125b-5p+高浓度组。对照组常规培养;模型组用OGD/R诱导建立细胞损伤模型;低浓度组、中浓度组、高浓度组分别加入2.5、5、10μmol/L利多卡因培养24 h后进行OGD/R处理;miR-NC+模型组、miR-125b-5p+模型组分别转染miR-NC、miR-125b-5p mimics后,进行OGD/R处理;anti-miR-NC+高浓度组、anti-miR-125b-5p+高浓度组分别转染anti-miR-NC、anti-miR-125b-5p加入浓度为10μmol/L利多卡因培养24 h,然后进行OGD/R处理。用MTT法检测细胞活力[吸光度值(A值)],流式细胞术检测细胞凋亡率,实时荧光定量PCR法检测miR-125b-5p表达,Western blotting法检测凋亡相关蛋白[剪切的半胱氨酸天冬氨酸蛋白水解酶3(Cleaved-caspase-3)、Bcl2关联X蛋白(Bax蛋白)]、磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路相关蛋白(p-PI3K、p-Akt蛋白)表达,用微板法、WST-1法及钼酸铵法检测氧化应激相关指标[丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)]。结果与对照组比较,模型组细胞活力低(P<0.05);与模型组比较,低浓度组、中浓度组、高浓度组细胞活力高(P均<0.05);不同浓度组细胞活力(A值)比较差异有统计学意义(P均<0.05)。与对照组比较,模型组细胞凋亡率,Cleaved-caspase-3、Bax相对表达量,MDA水平高(P均<0.05),SOD、CAT水平低(P均<0.05);与模型组比较,低浓度组、中浓度组、高浓度组细胞凋亡率,Cleaved-caspase-3、Bax蛋白相对表达量,MDA水平低,SOD、CAT水平高(P均<0.05);低浓度组、中浓度组、高浓度组细胞凋亡率,Cleaved-caspase-3、Bax蛋白相对表达量,MDA水平依次降低(P均<0.05),SOD、CAT水平依次升高(P均<0.05)。�
Objective To investigate the effect of lidocaine on oxygen-glucose deprivation/reoxygenation(OGD/R)-induced nerve cell damage and its mechanism of action.Methods Rat adrenal chromaffin cells(PC12 cells)were routinely cultured and randomly divided into the control group,model group,low-concentration group,medium-concentration group,high-concentration group,miR-NC+model group,microRNA-125b-5p(miR-125b-5p)+model group,anti miR-NC+high-concentration group,and anti-miR-125b-5p+high-concentration group,respectively.Cells in the control group underwent routine culture;cells in the model group were induced with OGD/R to establish cell injury models;cells in the low-concentration group,medium-concentration group,and high-concentration group were cultured with 2.5,5 and 10μmol/L lidocaine for 24 h,respectively,and then were treated with OGD/R.Cells in the miR-NC+model group and miR-125b-5p+model group were transfected with miR-NC and miR-125b-5p mimics,respectively,and then were treated with OGD/R.Cells in the anti-miR-NC+high-concentration group and the anti-miR-125b-5p+high-concentration group were transfected with anti-miR-NC and anti-miR-125b-5p,respectively,and then were added with 10μmol/L lidocaine for 24 h,and finally were treated with OGD/R.MTT assay was used to detect cell viability[absorbance value(A value)],flow cytometry was used to detect the apoptosis rate,and real-time fluorescence quantitative PCR was used to detect miR-125b-5p expression.Western blotting was used to detect the expression levels of apoptosis-related proteins[Cleaved caspase-3 and Bcl2 associated X protein(Bax protein)],phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)signaling pathway-related proteins(p-PI3K,p-Akt protein)],and microplate method,WST-1 method,and ammonium molybdate method were used to detect oxidative stress-related indicators[malondialdehyde(MDA),superoxide dismutase(SOD),and catalase(CAT)].Results Compared with the control group,the cell viability of the model group was lower(P<0.05).Compared with the model group
作者
杨文俊
许楠
董家瑞
黄天丰
李润林
YANG Wenjun;XU Nan;DONG Jiarui;HUANG Tianfeng;LI Runlin(Department of Anesthesiology,Jiangdu People's Hospital Affiliated to Medical College of Yangzhou University,Yangzhou 225200,China;不详)
出处
《山东医药》
CAS
2023年第31期38-42,71,共6页
Shandong Medical Journal
基金
国家自然科学基金资助项目(82001170)。
