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为了探究不同浓度尼古丁对体外细胞增殖分化的影响以及维生素C对尼古丁的生物学作用的影响,该文以兔成骨细胞为实验材料,对细胞增殖和各项分化指标进行了检测。MTT结果显示:与空白对照组相比,1×10-6,1×10-5mmol/L尼古丁组有促细胞增殖的作用,但是高浓度尼古丁(1 mmol/L)组对细胞增殖有明显的抑制作用。RT-PCR检测发现:用低浓度尼古丁处理细胞,ALP、COLI和OCN的基因表达上调;相反,高浓度尼古丁下调了细胞ALP、COLI和OCN的表达。ALP染色和Von Kossa钙结节染色也显示出高浓度尼古丁对成骨细胞的毒性作用。加入维生素C后,1 mmol/L尼古丁组对成骨细胞增殖和各基因表达的影响有所改善,类似的结果也见于ALP染色和Von Kossa染色。由此证实,极低浓度尼古丁对成骨细胞确有促进增殖、增强ALP活性和上调ALP、COLI、OCN基因表达的作用;但是,高浓度尼古丁却有相反的作用,抑制成骨细胞的增殖和分化。同时,维生素C具有部分拮抗高浓度尼古丁对成骨细胞毒性作用的能力。
In order to investigate the effect of different concentrations of nicotine on cell proliferation and differentiation in vitro and the effect of vitamin C on the biological effects of nicotine, the proliferation and differentiation of cells were examined using rabbit osteoblasts as experimental material. Compared with the blank control group, the MTT assay showed that the cell proliferation was inhibited in the 1 × 10-6 and 1 × 10-5mmol / L Nicotine groups, but the cell proliferation was significantly increased in the high concentration nicotine (1 mmol / L) group Inhibition. The results of RT-PCR showed that the gene expression of ALP, COLI and OCN was upregulated in cells treated with low concentration of nicotine. On the contrary, high concentration of nicotine down-regulated the expression of ALP, COLI and OCN. ALP staining and Von Kossa calcium nodule staining also showed toxic effects of high concentrations of nicotine on osteoblasts. The effect of 1 mmol / L nicotine on osteoblast proliferation and gene expression was improved after adding vitamin C. Similar results were found in ALP staining and Von Kossa staining. It was confirmed that very low concentrations of nicotine on osteoblasts do promote proliferation, increase ALP activity and upregulate ALP, COLI, OCN gene expression; However, the high concentration of nicotine has the opposite effect, inhibition of osteoblast proliferation and Differentiation. At the same time, vitamin C has the ability to partially antagonize the toxic effects of high concentrations of nicotine on osteoblasts.