【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。更多还原

【Abstract】 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.更多还原