补骨脂酚抑制PI3K/Akt/Nrf2/HO-1对氧糖剥夺所致SH-SY5Y细胞损伤的作用及机制  

Study on protective effect and mechanism of bakuchiol inhibiting PI3K/Akt/Nrf2/HO-1 pathway against oxygen-glucose deprivation-induced injury in SHSY5Y cells

作　　者：徐玥玮[1] 张珅 高晓明[1] 王丽[1] 杨满琴[1] 张梦翔[1] 杨晓丹 戴文玲[2] 蔡明[1] XU Yuewei;ZHANG Shen;GAO Xiaoming;WANG Li;YANG Manqin;ZHANG Mengxiang;YANG Xiaodan;DAI Wenling;CAI Ming(Acupuncture Hospital of Anhui University of TCM,Hefei 230061,China;China Pharmaceutical University,Nanjing 210000,China)

机构地区：[1]安徽中医药大学第二附属医院,安徽合肥230061 [2]中国药科大学,江苏南京210000

出　　处：《药物评价研究》2025年第2期448-454,共7页Drug Evaluation Research

基　　金：安徽省高等学校科学研究项目(自然科学类)重点项目(2022AH050524,2023AH050866);安徽中医药大学第二附属医院“杏林英才”培育计划项目(2023-0500-48-41,2023-0500-48-46);安徽省高校优秀青年教师项目(YQYB2024030)。

摘　　要：目的研究补骨脂酚对氧糖剥夺/复糖复氧(OGD/R)诱导人源性神经母细胞瘤细胞(SH-SY5Y)损伤的影响。方法MTT法检测补骨脂酚(5、10、20、40、80μmol·L^(-1))对SH-SY5Y细胞活性的影响。将SH-SY5Y细胞分为对照组、模型组、补骨脂酚(5、10、20μmol·L^(-1))组、补骨脂酚(20μmol·L^(-1))+ML385(2μmol·L^(-1),Nrf2抑制剂)组、补骨脂酚(20μmol·L^(-1))+LY294002[5μmol·L^(-1),磷脂酰肌醇3-激酶(PI3K)抑制剂]组。MTT法检测SH-SY5Y细胞活力,倒置显微镜观察各组SH-SY5Y细胞形态,试剂盒法检测细胞上清超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,实时荧光定量PCR(q RT-PCR)法和Western blotting法测定磷脂酰肌醇3-激酶(PI3K)、丝氨酸/苏氨酸激酶B(Akt)、核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)mRNA和蛋白水平。结果补骨脂酚对正常SH-SY5Y细胞存活率无影响。与对照组比较,模型组细胞存活率显著下降(P<0.01),细胞个数明显减少,部分细胞裂解成碎片,部分细胞死亡;SOD、CAT活性显著降低(P<0.01),PI3K/Akt/Nrf2/HO-1 mRNA及蛋白水平显著下调(P<0.05、0.01、0.001)。与模型组比较,补骨脂酚显著增加细胞存活率(P<0.01),明显增加细胞个数,显著上调SOD、CAT活性(P<0.05、0.01),显著上调PI3K/Akt/Nrf2/HO-1 mRNA及蛋白水平(P<0.05、0.01、0.001)。与补骨脂酚(20μmol·L^(-1))组比较,补骨脂酚+ML385可逆转补骨脂酚对HO-1 mRNA及蛋白的上调作用(P<0.05、0.01),补骨脂酚+LY294002可逆转补骨脂酚对Nrf2、HO-1 mRNA及蛋白水平的上调作用(P<0.05、0.01)。结论补骨脂酚可显著改善OGD/R诱导的SH-SY5Y细胞损伤,且机制可能是通过激活PI3K/Akt进而上调Nrf2/HO-1抑制氧化应激改善神经损伤。Objective To study the protected effect of bakuchiol on human neuroblastoma cells(SH-SY5Y)injury induced by oxygen glucose deprivation/reoxygenation(OGD/R).Methods The effect of BAK(5,10,20,40,and 80μmol·L^(-1))on the viability of SHSY5Y cells was detected by MTT assay.SH-SY5Y cells were divided into control group,model group,the BAK(5,10,20μmol·L^(-1))groups,the BAK(20μmol·L^(-1))+ML385(2μmol·L^(-1),Nrf2 inhibitor)group,and the BAK(20μmol·L^(-1))+LY294002[5μmol·L^(-1),phosphatidylinositol 3-kinase(PI3K)inhibitor]group.The viability of SH-SY5Y cells was detected by MTT assay,the morphology of SH-SY5Y cells in each group was observed by inverted microscope,the activities of superoxide dismutase(SOD)and catalase(CAT)in the cell supernatant were detected by kit method,and the mRNA and protein levels of PI3K,serine/threonine kinase B(Akt),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase 1(HO-1)were determined by real-time fluorescence quantitative PCR(qRT-PCR)and Western blotting.Results Bakuchiol had no effect on the survival rate of normal SH-SY5Y cells.Compared with the control group,the cell survival rate in the model group was significantly decreased(P<0.01),the number of cells was significantly reduced,some cells were lysed into fragments,and some cells died;the activities of SOD and CAT were significantly decreased(P<0.01),and the mRNA and protein levels of PI3K/Akt/Nrf2/HO-1 were significantly down-regulated(P<0.05,0.01,0.001).Compared with the model group,psoralen significantly increased the cell survival rate(P<0.01),significantly increased the number of cells,significantly up-regulated the activities of SOD and CAT(P<0.05,0.01),and significantly up-regulated the mRNA and protein levels of PI3K/Akt/Nrf2/HO-1(P<0.05,0.01,0.001).Compared with the BAK(20μmol·L^(-1))group,BAK+ML385 could reverse the upregulation effect of BAK on HO-1 mRNA and protein(P<0.05,0.01),and BAK+LY294002 could reverse the up-regulation effect of BAK on Nrf2 and HO-1 mRNA and protein levels(P<0.05,0.01)

关 键 词：补骨脂酚 SH-SY5Y细胞 脑缺血再灌注 氧糖剥夺/复糖复氧 Nrf2/HO-1 PI3K/AKT 

分 类 号：R285.5[医药卫生—中药学]