miR-146b-5p对脑缺血再灌注损伤细胞的影响

Effect of miR-146b-5p on cerebral ischemia-reperfusion injury cells

目的研究miR-146b-5p在脑缺血再灌注损伤(CIRI)中的作用和调控机制。方法建立神经元细胞PC12的糖-氧剥夺/再氧合(OGD/R)体外模型,并将细胞随机分为NC mimic组和miR-146b-5p mimic组。以细胞计数法检测细胞存活率;以酶联免疫吸附(ELISA)法检测肿瘤坏死因子-α(TNF-α)和白细胞介素-β(IL-β)含量;以试剂盒法检测活性氧(ROS)和超氧化物歧化酶(SOD)含量;以双荧光素酶报告实验验证miR-146b-5p和硫氧还蛋白互作蛋白(TXNIP)的结合关系;以蛋白质印迹法检测TXNIP蛋白表达。结果72 h时,NC mimic组和miR-146b-5p mimic组细胞存活率分别为(61.32±8.11)%和(81.45±9.34)%,TNF-α含量分别为(125.17±11.03)和(81.14±9.83)pg·m L^(-1),IL^(-1)β含量分别为(98.33±8.17)和(63.22±7.91)pg·m L^(-1),ROS含量分别为(45.67±3.98)和(35.20±3.01)U·m L^(-1),SOD含量分别为(30.11±2.99)和(51.08±6.75)U·m L^(-1),差异均有统计学意义(均P<0.05)。结论在脑缺血再灌注损伤中,miR-146b-5p靶向TXNIP促进细胞存活,抑制炎症反应和氧化应激改善脑缺血再灌注损伤。

ObjectiveTo explore the functions and regulatory mechanism of miR-146b-5p in cerebral ischemia-reperfusion injury(CIRI).Methods An in vitro model of oxygen-glucose deprivation/reoxygenation(OGD/R)of PC12 neurons was established,and the cells were randomly divided into two groups:NC mimic group and miR-146b-5p mimic group.Cell count kit(CCK)-8 assay was adopted to detect cell survival rate.Enzyme linked immunosorbent assay(ELISA)was performed to detect the content of tumor necrosis factor-α(TNF-α)and interlenkin-β(IL-β).The kit was used to detect the content of reactive oxygen species(ROS)and superoxide dismutase(SOD).Dual-luciferase reporter assay was used to verify the binding relationship between miR-146b-5p and TXNIP.Western blot was utilized to detect the expression of TXNIP protein.Results At 72 h,the cell survival rates of the NC mimic and miR-146b-5p mimic groups were(61.32±8.11)%and(81.45±9.34)%,the content of TNF-αwere(125.17±11.03)and(81.14±9.83)pg·m L^(-1),the content of IL^(-1)βwere(98.33±8.17)and(63.22±7.91)pg·m L^(-1);the content of ROS were(45.67±3.98)and(35.20±3.01)U·m L^(-1);the content of SOD were(30.11±2.99)and(51.08±6.75)U·m L^(-1),all with significant differences(all P<0.05).Conclusion In cerebral ischemia-reperfusion injury,miR-146b-5p targets TXNIP to promote cell survival,inhibit inflammation and oxidative stress,and improve cerebral ischemiareperfusion injury.