基于Nrf2/HO-1通路探讨葛根素对高糖诱导成骨细胞凋亡的影响  

Effect of Puerarin on the Apoptosis of Osteoblasts Induced by High Glucose Based on Nrf2/HO-1 Pathway

作　　者：洪荣华[1] 李永华[1] 张富国[1] Hong Ronghua;Li Yonghua;Zhang Fuguo(Department of Orthopaedics,Taizhou First People’s Hospital,Zhejiang Taizhou 318020,China)

机构地区：[1]台州市第一人民医院骨科,浙江台州318020

出　　处：《中国药师》2020年第5期804-809,共6页China Pharmacist

基　　金：台州市社会发展类一般项目(编号1701KY45)。

摘　　要：目的:探讨葛根素对高糖诱导的成骨细胞凋亡的影响,并探讨其可能机制。方法:体外培养小鼠成骨细胞MC3T3-E1细胞,试验分为对照组(葡萄糖浓度为5.5 mmol·L-1)、高糖组(葡萄糖浓度为33 mmol·L-1)、高糖+葛根素组(葛根素终浓度分别为0.1,1,10μmol·L-1)。采用CCK-8法检测葛根素对高糖诱导的MC3T3-E1细胞活力影响;采用流式细胞法观察葛根素对高糖诱导的MC3T3-E1细胞凋亡能力的影响;采用免疫印迹法(WB)检测凋亡相关蛋白以及核相关E2因子2/血红素加氧酶-1(Nrf2/HO-1)信号通路相关蛋白表达;添加Nrf2/HO-1信号通路特异性抑制剂ML385检测其对细胞凋亡的影响。结果:与对照组相比,高糖组细胞活力、B淋巴细胞瘤-2(Bcl-2)蛋白水平、HO-1蛋白水平及细胞核Nrf2蛋白水平降低(P<0.05),细胞凋亡率、Bcl-2相关X蛋白质(Bax)、cleaved-caspase 3蛋白水平及细胞浆Nrf2蛋白水平升高(P<0.05);与高糖组相比,高糖+各浓度葛根素组细胞活力、Bcl-2蛋白水平、HO-1蛋白水平及细胞核Nrf2蛋白水平依次升高(P<0.05),细胞凋亡率、Bax、cleaved-caspase 3蛋白水平及细胞浆Nrf2蛋白水平依次降低(P<0.05);与高糖+高浓度葛根素相比,高糖+高浓度葛根素+抑制剂组细胞凋亡率、Bax、cleaved-caspase 3蛋白表达升高(P<0.05),Bcl-2蛋白表达降低(P<0.05)。结论:葛根素可能通过激活Nrf2/HO-1信号通路,抑制高糖诱导的成骨细胞凋亡。Objective:To investigate the effect of puerarin on the apoptosis of osteoblasts induced by high glucose,and to explore the possible mechanism.Methods:Human osteoblasts MC3T3-E1 cells were cultured in vitro,and divided into the control group(5.5 mmol·L-1 of glucose),high glucose group(33 mmol·L-1 of glucose),high glucose+puerarin group(the final puerarin concentration was 0.1,1 and 10μmol·L-1,respectively).CCK-8 method was used to detect the effect of puerarin on the viability of MC3T3-E1 cells induced by high glucose;flow cytometry was used to observe the effect of puerarin on the apoptotic ability of MC3T3-E1 cells induced by high glucose;Western blotting(WB)was used to detect apoptotic-related proteins and Nrf2/HO-1 signal pathway-related proteins;the effect on the apoptosis was detected by adding Nrf2/HO-1 signaling pathway specific inhibitor ML385.Results:Compared to those in the control group,the cell viability,Bcl-2 protein level,HO-1 protein level and nucleus Nrf2 protein level in high glucose group all decreased(P<0.05),while the apoptotic rate,Bax,cleaved-caspase 3 protein level and plasma Nrf2 protein level increased(P<0.05).Compared to those in high glucose group,the cell viability,Bcl-2 protein level,HO-1 protein level and nucleus Nrf2 protein level in the high glucose+low puerarin group,high glucose+medium puerarin group and high glucose+high puerarin group all increased in turn(P<0.05),while the apoptotic rate,Bax,cleaved-caspase 3 protein level and plasma Nrf2 protein level decreased in turn(P<0.05).Compared with those in high glucose+high concentration puerarin group,the apoptotic rate,expression of Bax and cleaved-caspase 3 proteins of high glucose+high concentration puerarin+inhibitor group increased(P<0.05),and the expression of Bcl-2 protein decreased(P<0.05).Conclusion:Puerarin may inhibit the apoptosis of high glucose-induced osteoblasts by activating Nrf2/HO-1 signaling pathway.

关 键 词：葛根素 成骨细胞 高糖 凋亡 核相关因子2 血红素加氧酶 

分 类 号：R965.1[医药卫生—药理学]