去甲二氢愈创木酸类似物对过氧化氢诱导细胞损伤的保护作用  

Protective effect of nordihydroguaiaretic acid analogue on HO-induced cell damage

作　　者：王石健[1] 汪佳兵 WANG Shijian;WANG Jiabing(Department of Pharmacy,Taizhou Municipal Hospital,Taizhou318000,CHINA)

机构地区：[1]台州市立医院药剂科,浙江318000

出　　处：《江苏医药》2021年第10期977-980,F0003,共5页Jiangsu Medical Journal

基　　金：台州市科技计划项目(1901ky48)。

摘　　要：目的探讨去甲二氢愈创木酸(NDGA)类似物3g对过氧化氢(H_(2)O_(2))诱导细胞氧化应激损伤的作用。方法人神经母细胞瘤细胞系SH-SY5Y分为DMSO组(A组)、NDGA类似物3g 5μmol/L组(B组)、NDGA 5μmol/L组(C组)、H_(2)O_(2)组(D组)、NDGA类似物3g 5μmol/L+H_(2)O_(2)组(E组)。采用MTT法检测细胞生存率,4’,6-二脒基-2-苯基吲哚(DAPI)染色检测细胞凋亡情况,流式细胞术检测活性氧(ROS)水平,试剂盒检测丙二醛(MDA)水平,免疫荧光观察核因子E2相关因子2(Nrf2)入核情况,Western blot法检测Bax、Bcl-2、血红素氧化酶1(HO-1)和谷氨酸半胱氨酸连接酶催化亚基(GCLC)的蛋白表达。结果B组和C组细胞生存率较A组增加(P<0.05)。与A组相比,D组细胞发生凋亡现象,细胞生存率降低,MDA和ROS水平升高(P<0.05);而E组加入NDGA类似物3g后能逆转上述变化过程(P<0.05)。与A组相比,D组和E组Bax蛋白表达增加,Bcl-2蛋白表达减少(P<0.05),D组变化更加明显(P<0.05)。B组经NDGA类似物3g处理后能够诱导Nrf2进入细胞核。B组HO-1和GCLC蛋白表达较A组增加(P<0.05)。结论NDGA类似物3g通过降低ROS和MDA水平及细胞凋亡,并激活Nrf2通路,从而起到细胞保护作用。Objective To investigate the effect of nordihydroguaiaretic acid(NDGA)analogue 3 g on H_(2)O_(2)-induced cellular oxidative stress damage.Methods Human neuroblastoma cell line SH-SY5 Y was divided into five groups of A(DMSO negative control),B(NDGA analogue 3 g 5μmol/L),C(NDGA 5μmol/L),D(H_(2)O_(2)450μmol/L)and E(NDGA analogue 3 g 5μmol/L+H_(2)O_(2)450μmol/L).The cell survival and apoptosis were measured by MTT assay and DAPI staining,respectively.The level of reactive oxygen species(ROS)was determined by flow cytometry.The level of malondialdehyde was analyzed by the kit.The nuclear translocation of Nrf2 was observed by immunofluorescence assay.The protein expressions of Bax,Bcl-2,hemeoxygenase-1(HO-1)and glutamate-cysteine ligase catalytic subunit(GCLC)were detected by Western blot.Results The cell survival rate in groups of B and C was higher than that in group A(P<0.05).Compared with group A,the cell apoptosis occurred,the cell survival rate was decreased,and the levels of MDA and ROS were increased in group D(P<0.05),while which were significantly reversed in group E after the pretreatment of NDGA analogue 3 g(P<0.05).Compared with group A,the protein expression of Bax was increased,and the protein expression of Bcl-2 was decreased in groups of D and E(P<0.05),which changed more obviously in group D(P<0.05).The treatment of NDGA analogue 3 g could induce Nrf2 into the cell nucleus in group B.The protein expressions of HO-1 and GCLC in group B were higher than those in group A(P<0.05).Conclusion NDGA analogue 3 g plays a protective role in cell damage by reducing the levels of ROS and MDA,cell apoptosis and activating the Nrf2 pathway.

关 键 词：去甲二氢愈创木酸类似物 氧化损伤 细胞保护 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]