槲寄生多糖促进小鼠颅骨前骨细胞系MC3T3-E1的增殖及抑制凋亡的机制

赵磊 田守进 葛建飞 黄群 朱宏岩 董启榕

解剖学报 ›› 2019, Vol. 50 ›› Issue (4) : 459-464.

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解剖学报 ›› 2019, Vol. 50 ›› Issue (4) : 459-464. DOI: 10.16098/j.issn.0529-1356.2019.04.009
细胞和分子生物学

槲寄生多糖促进小鼠颅骨前骨细胞系MC3T3-E1的增殖及抑制凋亡的机制

  • 赵磊1 田守进1 葛建飞1 黄群1 朱宏岩2 董启榕3*
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Effect of mistletoe polysaccharose on mouse osteoblast cell line MC3T3-E1 promoting proliferation and  inhibiting apoptosis and its possible mechanism

  • ZHAO Lei1 TIAN Shou-jin1 GE Jian-fei1 HUANG Qun1 ZHU Hong-yan2 DONG Qi-rong 3*
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摘要

目的 检测槲寄生多糖对小鼠颅骨前骨细胞(MC3T3-E1)增殖、凋亡的影响并探讨其机制。 方法 提取槲寄生多糖,分别配置成不同质量浓度(0.625 g/L,1.25 g/L,2.5 g/L,5 g/L)作用MC3T3-E1小鼠颅骨前骨细胞及对照组。流式细胞术检测细胞周期及凋亡情况,用BrdU和碱性磷酸酶(ALP)试剂盒分别检测细胞增殖及细胞外碱性磷酸酶分泌变化,通过Real-time PCR检测成骨细胞相关基因mRNA的表达水平。 结果 与对照组相比,药物处理组随槲寄生多糖浓度的上升,细胞周期S期和G2/M的占比增多,在Annexin Ⅴ+/PI-象中的细胞百分数明显下降,细胞数量明显递增,细胞外ALP的活性呈明显递减,Runt相关转录因子2(Runtx2)、Ⅰ型胶原α1(COL1A1)链转录水平升高,且递增、递减效果都在2.5 g/L浓度之后趋于平稳。而骨钙素(OC)mRNA表达水平逐渐升高,在1.25 g/L之后平稳。 结论 槲寄生多糖能促进成骨细胞MC3T3-E1的增殖,抑制其凋亡,其机制可能与抑制碱性磷酸酶分泌以及促进骨钙素、Runt相关转录因子2、Ⅰ型胶原α1链的mRNA表达水平有关。

Abstract

Objective To observe the effect of mistletoe polysaccharose on proliferation and apoptosis of osteoblast and to explore the possible mechanism. Methods The mistletoe polysaccharose was extracted and made different concentrations (0.625 g/L, 1.25 g/L, 2.5 g/L, 5 g/L) to treat mouse skull bone cell line MC3T3-E1. Cell cycle and apoptosis were detected by flow cytometry. Cell proliferation and extracellular alkaline phosphatase secretion were detected by BrdU and alkaline phosphatase ELISA kits. The mRNA expression level of osteoblast related gene was detected by Real-time PCR. Results Compared with the control group, the result of treatment groups with increasing mistletoe polysaccharose concentrations were as follows: the proportions of S phase and G2/M phase in cell cycle increased and the cell count Annexin Ⅴ+/PI-significantly decreased, the number of cells increased, extracellular alkaline phosphatase activity obviously decreased. The increasing transcription levels of Runtrelated transcription factor 2(Runtx2) and collagen typeⅠ alpha 1(COL1A) were stabled over 2.5 g/L and the upregulation of mRNA expression of osteocalcin (OC) was steady at 1.25 g/L. Conclusion Mistletoe polysaccharose can promote the proliferation and inhibit the apoptosis of MC3T3-E1 cells, and its possible mechanism might be related to down-regulating of extracellular alkaline phosphatase activity and the grow exhanced expression of OC, Runtx2 and COL1A.

关键词

槲寄生多糖 / 小鼠颅骨前骨细胞 / 细胞周期 / 流式细胞术 / 小鼠

Key words

Mistletoe polysaccharose / MC3T3-E1 cell / Cell cycle / Flow cytometry / Mouse

引用本文

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赵磊 田守进 葛建飞 黄群 朱宏岩 董启榕. 槲寄生多糖促进小鼠颅骨前骨细胞系MC3T3-E1的增殖及抑制凋亡的机制[J]. 解剖学报. 2019, 50(4): 459-464 https://doi.org/10.16098/j.issn.0529-1356.2019.04.009
ZHAO Lei TIAN Shou-jin GE Jian-fei HUANG Qun ZHU Hong-yan DONG Qi-rong. Effect of mistletoe polysaccharose on mouse osteoblast cell line MC3T3-E1 promoting proliferation and  inhibiting apoptosis and its possible mechanism[J]. Acta Anatomica Sinica. 2019, 50(4): 459-464 https://doi.org/10.16098/j.issn.0529-1356.2019.04.009

参考文献

 [1]  Guo WQ, He PL. Research progress on animal models for prevention and treatment of osteoporosis by Chinese medicine [J]. China Journal of Traditional Chinese Medicine and Pharmacy, 2017,32 (2): 666-668. (in Chinese)
郭文清,何保丽.中医药防治骨质疏松症动物模型研究进展[J].中华中医药杂志,2017,32(2):666-668.
 [2] Gong ZhN,Zhang ShQ,Chen GX,et al.Structure and biological functions of the anticancer proteins from mistletoes[J].Chinese Journal of Biochemical Pharmaceutics, 2001, 22(5):259-262. (in Chinese)
龚祝南, 张双全, 陈国祥,等. 槲寄生类植物抗肿瘤活性蛋白成分的结构、功能及其作用机制的研究进展[J]. 中国生化药物杂志, 2001, 22(5):259-262.
 [3] Omeje EO, Khan MP, Osadebe PO, et al.Analysis of constituents of the eastern Nigeria mistletoe, Loranthus micranthus linn revealed presence of new classes of osteogenic compounds[J].J Ethnopharmacol,2014,151(1):643-651.
 [4] Wang J,Zhu YF.Extraction and separation of polysaccharides from mistletoe and its content determination[J].China Journal of Chinese Materia Medica,2007,32(22):2387-2390. (in Chinese)
王俊,朱一凡.槲寄生多糖的提取分离和含量测定[J].中国中药杂志,2007,32(22):2387-2390.
 [5] Ohtsu N, Hirano M, Arai H.Response of osteoblast-like MC3T3-E1 cells on bioactive titanium fabricated by a chemical treatment process using a calcium-phosphate slurry[J].J Biomed Mater Res A,2014,102(11):3838-3845.
 [6] Qin QS,Wang DM,Ma LT,et al.Characteristics of bone strength index and its relationship with body composition of Han nationality adults in Lanzhou City[J]. Acta Anatomica Sinica, 2016,47(6):838-842. (in Chinese)
秦青松,汪东梅,马路通,等.兰州市汉族成人骨强度指数特点及其与体成分的关系[J].解剖学报,2016,47(6):838-842.
 [7] Hannafon F, Cadogan MP.Recognition and treatment of postmenopausal osteoporosis[J].J Gerontol Nurs,2014,40(3):10-14. 
 [8] Zhang JL, Han X, Shan YJ,et al.Effect of bovine lactoferrin and human lactoferrin on the proliferative activity of the osteoblast cell line MC3T3-E1 in vitro[J].J Dairy Sci,2018,101(3):1827-1833.
 [9] Zhang X,Wang X,Gu BL, et al.Effects of Danshen on apoptosis and osteogenesis of MC3T3-E1 cells in high-iron environment[J].Chinese Journal of Experimental Surgery,2017,34(3):443-445. (in Chinese)
张晓,王啸,顾伯林,等.丹参对高铁环境下MC3T3-E1细胞凋亡、成骨功能的影响及其作用机制[J].中华实验外科杂志,2017,34(3):443-445.
 [10] Liu J, Zhang B, Song S,et al.Bovine collagen peptides compounds promote the proliferation and differentiation of MC3T3-E1 pre-osteoblasts[J].PLoS One,2014,9(6):e99920.
 [11] Yin WF, Li WK, Li GH, et al. Induction of osteogenic differentiation of cartilage stem cells by calcitonin gene-related peptide [J]. Chinese Journal of Physical Medicine and Rehabilitation, 2016, 38(5): 329-334. (in Chinese)
印卫锋,李文凯,李光辉, 等.降钙素基因相关肽诱导前软骨干细胞成骨分化的实验研究[J].中华物理医学与康复杂志,2016,38(5):329-334.
 [12] Hu HY,Liu CH.Signaling and transcription activator 1 gene silencing promote osteoblast differentiation in human periodontal ligament cells[J]. Stomatology,2016,36(10):884-889. (in Chinese)
呼海燕,刘彩宏.信号传导及转录激活因子1基因沉默促进人牙周膜细胞成骨细胞分化[J].口腔医学,2016,36(10):884-889.

基金

基金项目:张家港市科技局科技支撑项目(编号:ZKS1741)。

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