紫杉烷类化合物抗氧化应激作用与凋亡的关系

紫杉烷类化合物抗氧化应激作用与凋亡的关系
段瑞生;张慧平;谢涛;李燕;李俐涛;董玫;王维平
【期刊名称】《医药导报》
【年(卷),期】2013(32)4
【摘要】Objective To study the anti-oxidative effect of taxane. Methods Oxidative SK-N-SH cell model was established by treatment with suitable concentration of hydrogen peroxide. 7-Deacetyl-taxine B was extracted and purified from Taxus cuspidate, and used to treat the oxidative SK-N-SH cells. Cell viability was evaluated by using 3-(4,5)-dimethylthiahiazo (-z-yl )-3,5-di-phenytetrazoliumromide (MTT) assay. Apoptosis rates were detected using flow cytometry and the ultrastructure was observed by electron microscopy. Expression of Caspase-3 was analyzed using Western blotting. Results The viability of SK-N-SH cells was detected with the ratio of ( 99. 32 ±4. 30 ) % ( P>0. 05 ) after treated with 100 μmol· L-1 7-deacetyl-taxine B, and the viability of oxidative SK-N-SH cell model after treated with 1,10 and 100 μmol· L-1 7-deacetyl-taxine B were (25.90±
1.61)% ,(31.20±1. 25)% , (57. 6±3. 74)% (P<0. 05) ,respectively. After Annexin-V/PI staining the livability rate and apoptosis rate in negative control group were (50. 23±5. 31)% and (23. 27±1. 62)% , and the rates were (71. 20±3. 20)% , (19.20±
2. 11)% (P<0. 05) after treated with EGCG, and they were (79. 95±6. 13 )% ( P<0. 05) , (1
3. 09±1. 8)% (P< 0.05) after treated with 100 μmol· L-1 7-deacetyl-taxine B. In negative control group,
cells showed features of apoptosis, but after treated with 7-Deacetyl-taxine B the apoptosis and ultra-microstructure destruction were obviously alleviated. The caspase-3 protein expression of negative control group obviously decreased from (1. 30±0. 16) to (0. 77±0. 20) (P<0. 05) after treated with 7-deacetyl-taxine B. Conclusion The anti-oxidative activity of 7-Deacetyl-taxine B can decrease apoptosis.%目的探讨紫杉烷类化合物的抗氧化应激作用与细胞凋亡的关系.方法选择适当浓度过氧化氢处理神经母细胞瘤细胞(SK-N-SH)建立氧化应激细胞模型,用自红豆杉科植物(Taxus cuspidate)中分离纯化的紫杉烷类化合物7-Deacetyl-taxine B干预氧化应激模型细胞,用四甲基偶氮唑盐(MTT)微量酶反应比色法筛选具有神经保护活性的化合物,电镜观察其超微结构改变,流式细胞术观察其细胞凋亡率的变化,Western blot方法检测细胞Caspase-3表达水平的改变.结果正常对照组细胞经过浓度为100 μmol·L-1 7-Deacetyl-taxine B处理后,细胞活力为(99.32±4.30)％(P＞0.05),7-Deacetyl-taxine B化合物浓度1,10,100 μmol·L-1时氧化应激细胞活力分别为
(25.90±1.61)％,(31.20±1.25)％,(57.60±3.74)％(P＜0.05).Annexin-v/ PI染色后阴性对照组细胞生存率为(50.23±5.31)％,细胞模型凋亡率为
(23.27±1.62)％;EGCG处理后生存率为(71.20±3.20)％(P＜0.05),凋亡率为(19.20±2.11)％(P＜0.05)；7-Deacetyl-taxine B处理后生存率为(79.95±6.13)％(P＜0.05),凋亡率为(13.09±1.80)％(P＜0.05).阴性对照组超微结构呈凋亡改变,应用7-Deacetyl-taxine B后超微结构显示超微结构破坏减轻,凋亡减少,7-Deacetyl-taxine B干预后细胞内caspase-3蛋白表达水平,灰度值比值为(0.77±0.20),阴性对照组为(1.30±0.16),7-Deacetyl-taxine B组明显降低,差异有统计学意义(P＜0.05).结论 7-Deaceyl-taxine B可通过抗氧化应激起到抑制凋亡的作用.
【总页数】4页(P428-431)
【作者】段瑞生;张慧平;谢涛;李燕;李俐涛;董玫;王维平
【作者单位】河北医科大学第二附属医院神经内科,石家庄050000
【正文语种】中文
【中图分类】R741.05
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