华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

大蒜素对尼古丁致人牙周膜成纤维细胞氧化毒性的保护作用

李斌龙 谢晓莉 彭解英 罗小良 靳路远   

  1. 中南大学湘雅医院口腔内科, 长沙410008
  • 收稿日期:2011-02-25 修回日期:2011-02-25 出版日期:2011-02-20 发布日期:2011-02-20
  • 通讯作者: 谢晓莉,Tel:13973164578
  • 作者简介:李斌龙(1983—),男,山西人,硕士
  • 基金资助:

    湖南省科技厅基金资助项目(2010FJ3067);中南大学理科发展基金资助项目(1773-206001081)

Protective effect of allicin on human periodontal ligament cells with nicotine-induced oxidative damage

LI Bin-long, XIE Xiao-li, PENG Jie-ying, LUO Xiao-liang, JIN Lu-yuan   

  1. Dept. of Oral Medicine, Xiangya Hospital of Central South University, Changsha 410008, China
  • Received:2011-02-25 Revised:2011-02-25 Online:2011-02-20 Published:2011-02-20
  • Contact: XIE Xiao-li,Tel:13973164578

摘要:

目的研究大蒜素对尼古丁所致人牙周膜成纤维细胞(HPDLCs)氧化毒性的保护作用。方法1)建立尼古丁对HPDLCs氧化毒性的模型,采用水溶性四氮唑(WST)比色法选出可有效抑制HPDLCs存活率的尼古丁质量浓度X,用于后续实验。2)将体外培养的第5代HPDLCs分为空白组、尼古丁组、尼古丁+大蒜素组(大蒜素质量浓度分别为15、30、60 μg·mL-1),以不同培养液培养24 h后,采用WST比色法选出可显著改善HPDLCs存活率的大蒜素质量浓度Y用于后续实验。3)将体外培养的第5代HPDLCs分为空白组、尼古丁组、尼古丁+大蒜素组,分别于培养1、4、8、12、24 h后测量细胞内谷胱甘肽(GSH)浓度。结果0.8 mg·mL-1尼古丁即可显著抑制HPDLCs存活率(细胞存活率为空白组的77%,P<0.05,取X为0.8 mg·mL-1);而60 μg·mL-1大蒜素可显著改善此负性作用,将细胞存活率提升至尼古丁组的112%(P<0.05),且与空白组比较无统计学差异(P>0.05),取Y为60 μg·mL-1。在培养后的各时间点,尼古丁+大蒜素组GSH浓度与空白组相比均无统计学差异(P>0.05),但较尼古丁组均有明显提高(P<0.05);培养8 h时,尼古丁+大蒜素组GSH浓度与尼古丁组相比差异最大,前者较后者提高82%。结论60 μg·mL-1大蒜素可以显著降低尼古丁对HPDLCs的氧化损伤作用。

关键词: 人牙周膜成纤维细胞, 尼古丁, 氧化毒性, 大蒜素, 谷胱甘肽

Abstract:

Objective To explore the protective effect of allicin on nicotine-induced oxidative damage to human periodontal ligament cells(HPDLCs). Methods 1)Establish nicotine-induced oxidative damage model on HPDLCs. Use water-soluble tetrazolium(WST) colorimetric method to find out the nicotine concentration(X) that could inhibit HPDLCs’growth for the following experiments. 2)HPDLCs of the fifth passage were divided into 5 groups: The control group, the nicotine group and the nicotine+allicin groups(the concentration of allicin was 15, 30, and 60 μg·mL-1 respectively). Different kinds of culture media were added. Similarly, use WST colorimetric method to choose the allicin concentration(Y) that could significantly improve the survival rate of HPDLCs. 3)HPDLCs were divided into 3 groups: The control group, the nicotine group, the nicotine +allicin group and different media were added. The glutathion(GSH) concentrations in HPDLCs were determined in 1, 4, 8, 12 and 24 h respectively. Results 0.8 mg·mL-1 nicotine could inhibit the HPDLCs survival rate significantly(77% of the control, P<0.05). But 60 μg·mL-1 allicin could prevent the inhibition effects evidently, improving the survival rate to 112% of that of the nicotine group(P< 0.05) and reaching the survival rate level of control group(P >0.05). The GSH concentrations of nicotine +allicin group were higher than that of the nicotine group always(P<0.05) and by 82% at 8 h after culture, but had no difference with that of the control group(P>0.05). Conclusion 60 μg·mL-1 allicin can protect the HPDLCs against oxidative damage induced by nicotine.

Key words: human periodontal ligament cells, nicotine, oxidative damage, allicin, glutathion