摘要
目的探讨尼古丁抑制碘乙酸钠(MIA)诱导的骨关节炎软骨细胞凋亡。方法酶消化法分离大鼠原代软骨细胞,并用10-8,10-7,10-6,10-5mol/L尼古丁处理细胞48 h,随后实验分为5组,除正常组外,其余4组皆用4μM MIA处理24 h,并给予尼古丁。MTT法检测各组软骨细胞活力;Annexin V-FITC/PI流式双染细胞术检测各组软骨细胞凋亡;分光光度法检测各组软骨细胞中含半胱氨酸的天冬氨酸蛋白水解酶(Caspase 3)活性;Western blot分析磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)激活状况及下游靶分子Bax,Bcl-2的表达情况。结果 10-7,10-6mol/L尼古丁显著促进大鼠软骨细胞活力(P<0.05),10-5mol/L尼古丁显著降低大鼠软骨细胞活力(P<0.05),10-8mol/L尼古丁对大鼠软骨细胞活力无影响(P>0.05)。10-8,10-7,10-6mol/L尼古丁能剂量依赖性的提高MIA诱导的大鼠软骨细胞活力,并抑制MIA诱导的大鼠软骨细胞凋亡及Caspase 3活性(P<0.05);10-7,10-6mol/L尼古丁能提高PI3K表达及AKT磷酸化水平,并下调促Bax表达,上调Bcl-2表达(P<0.05)。结论一定剂量尼古丁能显著的抑制MIA诱导的大鼠软骨细胞凋亡,可能与PI3K/AKT信号通路有关。
Objective To explore inhibition of nicotine on apoptosis of chondrocytes induced by monosodium iodoacetate（ MIA）. Methods Rat primary chondrocytes were isolated by enzyme digestion,and the cells were treated with 10- 8,10- 7,10- 6,10- 5mol / L nicotine for 48 h. The cases were randomly divided into five groups,except for normal group,the other four groups were treated with 4 μmol / L MIA 24 h,and three groups were treated 10- 8,10- 7,10- 6mol /L nicotine. The viability of chondrocytes was detected by MTT assay. The apoptosis of chondrocytes was examed by Annexin V-FITC / PI flow dual-staining method. The activity of cysteinyl aspartate specific proteinase 3（ Caspase 3） was measured by spectrophotography method. The activation of phosphatidylinositol 3 kinase（ PI3K） / protein kinase B（ AKT）and the expression of down-stream molecule Bax,Bcl-2 was assayed by western blot. Results 10- 7,10- 6mol / L nicotine increased chondrocytes＇ viability（ P 〈 0. 05）,10- 5mol / L nicotine reduced chondrocytes＇ viability（ P 〈 0. 05）,and 10- 8mol / L nicotine didn‘t effect on chondrocytes＇ viability（ P 〉0. 05）. 10- 8,10- 7,10- 6mol / L nicotine could increaseMIA-induced chondrocytes＇ viability（ P 〈 0. 05）,suppress MIA-induced chondrocytes＇ apoptosis and the activity of MIAinduced Caspase 3（ P 〈 0. 05）. Moreover,10- 7,10- 6mol / L nicotine could increase the expression of PI3 K and phosphorylation of AKT（ P 〈 0. 05）,down-regulate the expression of Bax and up-regulate the expression of Bcl-2 in MIAinduced rat chondrocytes（ P 〈 0. 05）. Conclusion These results suggested nicotine could exert anti-apoptosis in MIAinduced rat chondrocytes,which might be related to PI3 K / AKT signal pathway.
出处
《中国比较医学杂志》
CAS
北大核心
2016年第3期40-45,共6页
Chinese Journal of Comparative Medicine
基金
海南省卫生厅基金资助项目(琼卫-2013资助-036号)
关键词
尼古丁
骨关节炎
软骨细胞
凋亡
Nicotine
Ostearthritis
Chondrocytes
Apoptosis
作者简介
韩贵宾（1974-），男，硕士，副主任医师，研究方向：关节外科。E-mail：hanguibin456@163．com。
[通讯作者]张寿，Email：shouzhang456@163．com.
