银杏二萜内酯通过拮抗PAFR和调节SIRT1/STAT3抑制氧化应激诱导的PC12神经元衰老研究  

Ginkgo diterpene lactones inhibit oxidative stress-induced PC12 neuronal aging by antagonizing PAFR and regulating SIRT1/STAT3

作　　者：杜雨芯 操娇娇 张倩霞 杨颖博[2,3] 吴伟 肖保国 肖伟 DU Yu-xin;CAO Jiao-jiao;ZHANG Qian-xia;YANG Ying-bo;WU Wei;XIAO Bao-guo;XIAO Wei(School of Pharmacy,Nanjing University of Traditional Chinese Medicine,Nanjing 210046,China;Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Jiangsu Kanion Pharmaceutical Co.,Ltd.,Lianyungang 222001,China;Institute of Neurology,Huashan Hospital Affiliated to Fudan University,Shanghai 200040,China)

机构地区：[1]南京中医药大学药学院,江苏南京210046 [2]上海中医药大学中药研究所,上海201203 [3]江苏康缘药股份有限公司,江苏连云港222001 [4]复旦大学附属华山医院神经病学研究所,上海200040

出　　处：《中草药》2023年第9期2793-2801,共9页Chinese Traditional and Herbal Drugs

基　　金：2021年国家中医药管理局青年岐黄学者项目。

摘　　要：目的探讨银杏二萜内酯(ginkgo diterpene lactone,GDL)对氧化应激诱导的细胞衰老的保护作用及机制。方法建立H_(2)O_(2)诱导PC12神经元衰老细胞模型,分别给予血小板活化因子拮抗剂WEB-2086或GDL预处理,采用CCK-8试剂盒检测细胞活性;β-半乳糖甘酶染色法检测衰老细胞率;DAPI染色法观察细胞平均核体积;Western blotting检测p21、p53、核纤层蛋白B1(Lamin B1)、血小板活化因子受体(platelet activating factor receptor,PAFR)、沉默调节蛋白1(Sirtuin 1,SIRT1)、磷酸化信号转导和转录激活蛋白3(phosphorylated signal transducer and activator of transcription 3,p-STAT3)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、神经营养因子3(neurotrophin 3,NT3)和神经生长因子(nerve growth factor,NGF)蛋白表达;TUNEL染色法观察细胞凋亡。结果与对照组比较,模型组显著降低了细胞的活性(P<0.05),衰老细胞数显著增加(P<0.01),PAFR表达显著升高(P<0.001)。与模型组比较,WEB-2086显著抑制PAFR表达(P<0.001),显著升高SIRT1蛋白表达(P<0.05),显著减少β-半乳糖甘酶衰老细胞率(P<0.05);GDL显著升高了细胞活力(P<0.05),显著上调Lamin B1、SIRT1、BDNF、NT3的表达(P<0.05、0.01),显著下调PAFR、p-STAT3、p21、p53蛋白表达(P<0.05、0.01)。结论GDL可以保护神经元并抑制细胞凋亡,延缓H_(2)O_(2)代表的氧化应激诱导PC12神经元的衰老,其机制可能与拮抗PAFR、调节SIRT1/STAT3信号通路有关。Objective To explore the protective effect and mechanism of ginkgo diterpene lactone(GDL)on cell senescence induced by oxidative stress.Methods The cell model of PC12 neuron aging induced by H_(2)O_(2)was established,and platelet activating factor antagonist WEB-2086 or GDL was pretreated respectively,cell activity was detected by CCK-8 kit;β-Galactosidase staining was used to detect the aging cell rate;The average nuclear volume of cells was observed by DAPI staining;p21,p53,Lamin B1,platelet activating factor receptor(PAFR),Sirtuin 1(SIRT1),phosphorylated signal transducer and activator of transcription 3(p-STAT3),brain-derived neurotrophic factor(BDNF)and neurotrophic factor 3(NT3)and nerve growth factor(NGF)protein expressions were detected by Western blotting;Apoptosis was observed by TUNEL staining.Results Compared with control group,the activity of cells in model group was significantly decreased(P<0.05),the number of aging cells was significantly increased(P<0.01)and the expression of PAFR was significantly increased(P<0.001).Compared with model group,WEB-2086 significantly inhibited the expression of PAFR(P<0.001),significantly increased the expression of SIRT1 protein(P<0.05),and significantly reduced the aging cell rate ofβ-galactosidase(P<0.05).GDL significantly increased the cell viability(P<0.05),significantly increased the expressions of Lamin B1,SIRT1,BDNF and NT3(P<0.05,0.01),and significantly decreased the expressions of PAFR,p-STAT3,p21 and p53(P<0.05,0.01).Conclusion GDL can protect neurons,inhibit apoptosis and delay the aging of PC12 neurons induced by oxidative stress represented by H_(2)O_(2),and its mechanism may be related to antagonizing PAFR and regulating SIRT1/STAT3 signaling pathway.

关 键 词：银杏二萜内酯 血小板活化因子受体 应激诱导的早衰 SIRT1/STAT3信号通路 凋亡 

分 类 号：R285.5[医药卫生—中药学]