尼古丁对腰椎终板软骨细胞增殖及AQP1表达调控的影响

Effect of nicotine on expression of AQP1 and proliferation of endplate chondrocytes

目的探讨尼古丁对腰椎终板骨细胞增殖的影响,了解尼古丁对水通道蛋白1(Aquaporin 1,AQP1)mRNA和蛋白表达的调控作用。方法培养大鼠腰椎终板软骨细胞,置于培养板上,实验组加入0.025、0.05、0.1、0.2 g/L系列浓度的尼古丁硫酸盐,对照组只加入培养液,观察不同浓度的尼古丁对终板软骨细胞生物活性的影响及其对AQP1蛋白及mRNA表达的影响。结果尼古丁浓度为0.05 g/L时,对终板软骨细胞的增殖即有显著的抑制作用。当尼古丁的浓度为0.025 g/L时,在蛋白和mRNA水平,其对终板软骨细胞AQP1的表达无明显抑制作用(P>0.05);当其浓度为0.05 g/L及以上时,与对照组相比,对终板软骨细胞AQP1的表达有显著的抑制作用(P<0.05)。结论吸烟对软骨细胞的活性有抑制作用,可以下调AQP1的表达,其对AQP1的调控作用和对终板软骨细胞活性的影响可能是其作为椎间盘退变高风险因素的原因之一。

Objective To investigate the effect of nicotine on the expression of AQP1 and the proliferation of endplate chon- droeytes. Methods Rat lumbar spinal endplate chondrocytes were cultivated. By Western blotting and reverse transcription polymer- ase chain reaction, the different concentrations of nicotine＇s effects on the expression of AQP1 was assayed, and the bioactivity of end- plate chondrocytes was evaluated by MTT. Results Nicotine inhibited the proliferation of endplate chondrocytes at 0.05 g/L level. At 0. 025 g/L level, nicotine had no effect on the protein and mRNA expression of AQP1 ; but at 0.05 g/L level, it inhibited the protein and mRNA expression of AQP1 significantly, compared with the control group（ P 〈 0.05 ） ; at 0.1 g/L and 0.2 g/L level, the effect of inhibition was more significant（P 〈 0.01 ）. Conclusions The high risk factors of intervertebral disc degeneration, such as smoking, had significant negative effect on the expression of AQP1. Its modulations of AQP1 may be one of reasons of causing the degeneration of intervertebral discs.