β-细辛醚对缺氧损伤的PC12细胞NMDAR亚基表达的影响  

Effects of β-Asarone on Expression of NMDAR Subunits in Hypoxia Injured PC12 Cells

作　　者：马进 李振万 江湧[1] MA Jin;LI Zhenwan;JIANG Yong(The First Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405

出　　处：《中药新药与临床药理》2020年第9期1015-1020,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金项目(04300378)。

摘　　要：目的观察石菖蒲挥发油主要成分β-细辛醚对缺氧损伤的PC12细胞N-甲基-D-天冬氨酸受体(Nmethyl-D-aspartate receptor,NMDAR)亚基mRNA及蛋白表达的影响。方法将细胞分为正常组、模型组和治疗组。无血清培养基培养细胞为正常组,10 mmol·L^-1Na2S2SO4无血清培养基培养细胞为模型组,1μmol·L^-1β-细辛醚+10 mmol·L^-1Na2S2SO4无血清培养基培养细胞为治疗组,各组均处理1 h。倒置显微镜观察细胞形态,CCK-8法检测细胞活性,流式细胞术检测细胞凋亡率,Q-PCR法检测NMDAR亚基mRNA表达,Western Blot法检测NR1、NR2A、NR2B蛋白表达。结果与模型组比较,治疗组细胞形态呈现明显差异;细胞活性明显升高(P<0.01);凋亡率明显降低(P<0.01);NR2B mRNA表达明显升高(P<0.05),其他亚基mRNA表达的差异无统计学意义(P>0.05),扩增未得到NR3B基因的产物;NR1蛋白表达的差异无统计学意义(P>0.05),未检测到NR2A、NR2B蛋白表达。结论β-细辛醚对PC12细胞缺氧损伤的保护作用可能与NR2B mRNA表达升高有关。Objective To observe effects of β-asarone on expression of N-methyl-D-aspartate receptor(NMDAR)subunits mRNA and protein in hypoxia-injured PC 12 cells.Methods Cells were divided into normal,model and treatment groups.Hypoxia model was established by treating cells with 10 mmol·L^-1 Na2 S2 SO4.The treatment group was treated with 1 μmol·L^-1 β-asarone+10 mmol·L^-1 Na2 S2 SO4,this process took a total of 1 h.Cell morphology was observed by inverted microscope,cell activity was detected by CCK-8 method,apoptosis rate was detected by flow cytometer,NMDAR subunit mRNA expression was detected by Q-PCR,NR1 and NR2 A-B protein expression were detected by Western Blot.Results Compared with the model group,the cells of the treatment group had smaller cell shrinkage,better transparency and higher cell density.Compared with the model group,the intervention of β-asarone can improve cell viability(P<0.01),reduce hypoxia-induced PC 12 cell apoptosis(P<0.01),and increase NR2 B mRNA expression(P<0.05);but it has no significant effect on the mRNA expression of other subunits(P>0.05).And there is no significant effect on NR1 protein expression(O> 0.05).NR2 A and NR2 B protein expressions were not detected.Conclusion The protective mechanism of β-asarone against hypoxia-injury in PC 12 cells may be related to the increase of mRNA expression of NR2 B.

关 键 词：石菖蒲 Β-细辛醚 PC12细胞 缺氧损伤 NMDAR亚基 NR2B 

分 类 号：R285.5[医药卫生—中药学]