N-乙酰半胱氨酸对脂多糖诱导的成骨细胞相关基因及蛋白表达影响的研究

doi:10.13701/j.cnki.kqyxyj.2017.11.005

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (11): 1151-1155.DOI: 10.13701/j.cnki.kqyxyj.2017.11.005

N-乙酰半胱氨酸对脂多糖诱导的成骨细胞相关基因及蛋白表达影响的研究

张慧, 李汪阳, 黄英, 蔡跃, 郭玲^*

西南医科大学附属口腔医院修复科四川泸州 646000

收稿日期:2017-04-29 出版日期:2017-11-20 发布日期:2017-11-29
通讯作者: 郭玲,E-mail:gl2005202@foxmail.com
作者简介:张慧(1992~ ),女,四川省凉山州人,硕士在读,主要从事口腔种植修复的研究工作。
基金资助:
四川省科技厅面上项目(编号:2017JY0111)
泸州市-校联合(编号:2016LZXNYD-J20)

Effect of N-acetyl Cysteine on Lipopolysaccharide Induced Expressions of Genes and Proteins in MC3T3-E1 Osteoblasts

ZHANG Hui, LI Wang-yang, HUANG Ying, CAI Yue, GUO Ling^*

Department of Prosthodontics, Stomatological Hospital of Southwest Medical University, Luzhou 646000, China

Received:2017-04-29 Online:2017-11-20 Published:2017-11-29

摘要/Abstract

摘要： 目的:研究N-乙酰半胱氨酸(NAC)对脂多糖(LPS)诱导的成骨细胞相关基因及蛋白表达的影响,为临床运用N-乙酰半胱氨酸预防和治疗种植体周围炎导致的种植体松动和脱落提供理论基础。方法:采用体外培养小鼠成骨细胞系(MC3T3-E1),以不同浓度N-乙酰半胱氨酸及脂多糖刺激细胞,分别在作用(6 h、24 h、48 h、72 h)后用cck-8法检测不同浓度N-乙酰半胱氨酸及脂多糖对细胞增殖情况的影响;将MC3T3-E1细胞随机分为4组:空白对照组、LPS( 10 mg/L LPS)组、NAC(1 mmol/L)组、NAC+LPS组(NAC预刺激30 min后,NAC与LPS共同作用),实时荧光定量PCR法及ELISA法分别检测MC3T3-E1细胞碱性磷酸酶(ALP)、骨钙素(BGP)、白介素6(IL-6)及核转录因子(NF-kB)基因及蛋白表达的情况。结果:随着NAC浓度的提高,MC3T3-E1的增殖率逐渐增加,在1 mmol/L时,细胞的增殖达到最大,但随着NAC浓度的继续增加,MC3T3-E1细胞的增殖率反而降低,表明高浓度的NAC具有毒性作用。NAC能拮抗LPS对MC3T3-E1细胞ALP和 BGPmRNA水平的下调作用,同时拮抗LPS对MC3T3-E1细胞ALP和 BGP蛋白表达的抑制作用;NAC能够抑制LPS对MC3T3-E1细胞IL-6和NF-kB mRNA水平的上调作用,同时降低LPS诱导的MC3T3-E1细胞IL-6和NF-kB蛋白的表达水平。结论:NAC可促进LPS诱导的MC3T3-E1细胞ALP、BGP表达,同时NAC通过抑制NF-kB活性,从而抑制LPS诱导的MC3T3-E1细胞表达IL-6等炎症因子。

关键词: N-乙酰半胱氨酸, 脂多糖, 成骨细胞, NF-kB, IL-6

Abstract: Objective: To evaluate the effect of N-acetyl cysteine on Lipopolysaccharide induced expressions of genes and proteins in MC3T3-E1 osteoblasts. Methods: The osteoblasts were isolated from the calvarias of mice and cultured in a-MEM. Cells were stimulated with various concentrations of NAC or LPS and then the proliferation of the cells was observed at different time points (6h, 24h, 48h, and 72h) by CCK-8. The expression of ALP, BGP, NF-KB, and IL-6 were determined by qPCR and ELISA analyses. Results: The proliferation of the cells increased with the increasing concentrations of NAC. When the concentration of NAC was 1mmol/L, the proliferation of the cells reached the maximum. A higher concentration of NAC had toxic effects. The down-regulated gene and protein expressions of ALP and BGP induced by 10μg/ml LPS were promoted significantly after treated with 1mmol/L NAC. The up-regulated gene and protein expressions of IL-6 and NF-kB induced by 10μg/ml LPS were decreased significantly after treated with 1mmol/L NAC. Conclusion: Expressions of ALP and BGP induced by LPS were promoted significantly after treated with NAC. NAC could decrease the expression of IL-6 induced by LPS through the NF-kB signaling.

Key words: N-acetyl cysteine, Lipopolysaccharide, MC3T3-E1 osteoblast, NF-kB, IL-6

中图分类号:

R329.2

张慧, 李汪阳, 黄英, 蔡跃, 郭玲. N-乙酰半胱氨酸对脂多糖诱导的成骨细胞相关基因及蛋白表达影响的研究[J]. 口腔医学研究, 2017, 33(11): 1151-1155.

ZHANG Hui, LI Wang-yang, HUANG Ying, CAI Yue, GUO Ling. Effect of N-acetyl Cysteine on Lipopolysaccharide Induced Expressions of Genes and Proteins in MC3T3-E1 Osteoblasts[J]. Journal of Oral Science Research, 2017, 33(11): 1151-1155.

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N-乙酰半胱氨酸对脂多糖诱导的成骨细胞相关基因及蛋白表达影响的研究

Effect of N-acetyl Cysteine on Lipopolysaccharide Induced Expressions of Genes and Proteins in MC3T3-E1 Osteoblasts

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