木犀草苷对阿尔茨海默病模型细胞凋亡和炎性因子表达的研究  

Study of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer’s disease

作　　者：王翠[1] 杨畅[1] 金玉 高蜜[1] 张雯[1] 王琼[2] 金海涛[1] WANG Cui;YANG Chang;JIN Yu;GAO Mi;ZHANG Wen;WANG Qiong;JIN Haitao(Department of Encephalopathy,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430000,China;Department of Endocrinology,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430000,China)

机构地区：[1]武汉市中医医院脑病科,430000 [2]武汉市中医医院内分泌科,430000

出　　处：《天津医药》2023年第7期701-706,共6页Tianjin Medical Journal

基　　金：武汉市中医药科研项目(WZ22C65)。

摘　　要：目的探究木犀草苷通过下调有丝分裂原活化蛋白激酶激酶激酶3(MEKK3)对阿尔茨海默病(AD)模型细胞凋亡和炎性因子表达的影响及可能机制。方法体外培养PC12细胞,经不同剂量(6.25、12.5、25 mg/L)木犀草苷孵育24 h后,建立β淀粉样肽(Aβ)_(25-35)诱导的AD细胞模型;另转染MEKK3小干扰RNA或过表达载体至PC12细胞,经25 mg/L木犀草苷孵育24 h后,建立Aβ_(25-35)诱导的AD细胞模型。细胞计数试剂盒-8(CCK-8)检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,酶联免疫吸附试验(ELISA)检测炎性因子[肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1β]水平,蛋白免疫印迹法(Western blot)检测MEKK3蛋白表达。结果木犀草苷可降低Aβ_(25-35)诱导的PC12细胞增殖抑制率、凋亡率、炎性因子(TNF-α、IL-6、IL-1β)水平,且降低细胞中MEKK3蛋白表达(P<0.05);敲减MEKK3可降低Aβ_(25-35)诱导的PC12细胞增殖抑制率、凋亡率、炎性因子(TNF-α、IL-6、IL-1β)水平,过表达MEKK3呈相反作用;上调MEKK3表达可减弱木犀草苷对Aβ_(25-35)诱导的PC12细胞增殖抑制率、凋亡率和炎性因子水平的影响。结论木犀草苷可能通过下调MEKK3抑制AD模型细胞凋亡及炎性因子水平,具有治疗AD的潜在价值。Objective To explore the effect and possible mechanism of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer′s disease(AD)by down-regulating mitogen-activated protein kinase kinase kinase 3(MEKK3).Methods PC12 cells were cultured in vitro and incubated with different doses(6.25 mg/L,12.5 mg/L,25 mg/L)of cynaroside for 24 h to establish an AD cell model induced by Aβ_(25-35).In addition,MEKK3 small interfering RNA or overexpression vector was transfected into PC12 cells,and Aβ_(25-35) induced AD cell model was established after incubation with 25 mg/L luteolin for 24 h.CCK-8 was used to detect inhibition rate of cell proliferation,and flow cytometry was used to detect apoptosis rate.ELISA was used to detect inflammatory factors(TNF-α,IL-6,IL-1β).The protein expression of MEKK3 was detected by Western blot assay.Results Cynaroside could decrease the proliferation inhibition rate,apoptosis rate,inflammatory cytokines(TNF-α,IL-6,IL-1β)and MEKK3 protein expression in PC12 cells induced by Aβ_(25-35)(P<0.05).Knockdown of MEKK3 could reduce the proliferation inhibition rate,apoptosis rate and inflammatory cytokines(TNF-α,IL-6,IL-1β)of Aβ_(25-35)-induced PC12 cells,while overexpression of MEKK3 showed the opposite effect.Upregulation of MEKK3 expression attenuated effects of cynaroside on the proliferation inhibition rate,apoptosis rate and inflammatory factors of Aβ_(25-35)-induced PC12 cells.Conclusion Cynaroside may inhibit apoptosis and expression of inflammatory factors of AD model cells by down-regulating MEKK3,which has potential value in the treatment of AD.

关 键 词：木犀草苷 阿尔茨海默病 PC12细胞 MAP激酶激酶激酶类 细胞凋亡 炎性因子 

分 类 号：R742[医药卫生—神经病学与精神病学]