[1]彭磊,程少文,沈跃,等.二氮嗪对软骨细胞氧化损伤的影响[J].中医正骨,2014,26(05):3-8.
　Peng Lei*,Cheng Saowen,Shen Yue,et al.Diazoxide influence on cartilage cells from oxidative damage[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2014,26(05):3-8.
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二氮嗪对软骨细胞氧化损伤的影响()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第26卷
期数:
2014年05期
页码:
3-8
栏目:
基础研究
出版日期:
2014-05-31

文章信息/Info

Title:
Diazoxide influence on cartilage cells from oxidative damage
作者:
彭磊1; 程少文1; 沈跃2; 应晓洲2; 赵志蓉2; 安涛2; 周英勇2; 程晓杰2; 陈庆玉2; 吕传柱3
1.海南医学院附属医院,海南 海口 570100; 2.温州医科大学,浙江 温州 325000; 3.海南医学院,海南 海口 570100
Author(s):
Peng Lei*;  Cheng Saowen;  Shen Yue;  Ying Xiaozhou;  Zhao Zhirong;  An Tao;  Zhou Yingyong;  Cheng Xiaojie;  Chen Qingyu;  Lu Chuanzhu.*
The affiliated hospital to Hainan medical colledge, Haikou, 570100,Hainan,China
关键词:
二氮嗪 软骨细胞 活性氧 凋亡
Keywords:
Diazoxide;  Chondrocytes;  Reactive oxygen species;  Apoptosis
摘要:
目的:探讨二氮嗪对双氧水诱导的软骨细胞氧化损伤的影响。方法:取SD乳鼠膝关节软骨细胞进行原代培养,将对数生长期的软骨细胞分成5组,分别为阴性对照组(A组),双氧水组(B组),H2O2+0.1 μmol·L-1二氮嗪组(C组),H2O2+1.0 μmol·L-1二氮嗪组(D组),H2O2+10 μmol·L-1二氮嗪组(E组); A组细胞不做特殊处理,B组用0.3 mmol·L-1双氧水在37 ℃恒温箱内孵育4 h,C、D、E、F组预先分别用0.1 μmol·L-1 DZ, 1.0 μmol·L-1DZ,10 μmol·L-1DZ,100 μmol·L-1在37℃的恒温箱孵育30分钟,PBS洗涤细胞,再用0.3 mmol·L-1双氧水在37℃恒温箱内孵育4小时。分别检测ABCDE各组细胞的细胞活性,ROS的量,细胞凋亡情况。结果:①MTT法检测各组细胞活性,细胞活性率从大至小依次为D组>C组>E组>F组>B组; ②用活性氧探针DCFH-DA检测各组细胞中的ROS的量,ROS产量从多至少依次为B组>E组>D组>C组>A组; ③流式细胞仪检测各组细胞凋亡情况,各组细胞凋亡率由大至小依次为B组>E组>D组>C组。④Western bolt检测各组软骨细胞中HIF-1α蛋白的表达,各组软骨细胞中HIF-1α蛋白表达量依次为C组>D组>E组>A组>B组。结论:二氮嗪可降低双氧水诱导的软骨细胞的凋亡率,同时也降低双氧水诱导的软骨细胞ROS的产生,并诱导软骨细胞产生HIF-1α蛋白,可能存在二氮嗪诱导HIF-1α的表达,引起下游相关基因的转录,总体提高软骨细胞对氧化损伤的耐受力,阻碍自由基诱导的软骨细胞的凋亡。
Abstract:
Objective:To investigate diazoxide infection in hydrogen peroxide induced cartilage cells.Method:We obtain the carilage cells from the SD neonatal rat articular cartilages. The cartilage cells were divided into five groups,namely the control group(A), the hydrogen peroxide group(B),H2O2 + 0.1 μmol·L-1DZ group(C), H2O2 +1.0 μmol·L-1 DZ group(D), H2O2 +10 μmol·L-1 DZ group(E); group A had no special treatment,group B incubated with 0.3 mmol/L hydrogen peroxide for 4 hours in 37 ℃ incubator, the C, D, E group were preincubated with 0.1 μmol·L-1DZ 1.0 μmol·L-1DZ,10 μmol·L-1 DZ in a 37 ℃ incubator incubated 30 minutes, then incubated with 0.3 mmol·L-1 hydrogen peroxide solution in 37 ℃ incubator.The amount of the ROS and the apoptosis rate of cells in each group were detected.Results:①.Using the MTT assay, the order of the rate of cells activity: group A> group D > group C > group E >group B. ②. The ROS productions were detected by the DCFH-DA, the order of ROS production as follow: group B> group E> group D> group C> group A. ③. The apoptosis rates of cells were detected by Flow cytometer.The order of apoptosis rates of cartilage cells as follow: group B> group E> group D> group C group.④. the expression of HIF-1α in each chondrocytes group detected by western blot,as follow: group C> group D > group E > group A > group B.Conclusion:Diazoxide can reduce the apoptosis rates of hydrogen peroxide-induced chondrocytes, while also reducing the ROS production of the hydrogen peroxide-induced chondrocyte.It can be inferred that,diazoxide may reduce the apoptosis rates and mortality of chondrocyte by reducing the ROS production of chondrocyte and increasing the expression of HIF-1α protein.

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更新日期/Last Update: 1900-01-01