葛根素对大鼠下颌骨骨髓源巨噬细胞破骨细胞向分化的影响

doi:10.19438/j.cjoms.2019.03.001

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (3): 193-197.doi: 10.19438/j.cjoms.2019.03.001

葛根素对大鼠下颌骨骨髓源巨噬细胞破骨细胞向分化的影响

马旭辉¹, 张鹏², 杨屹羚³, 徐弘远³, 龚心怡³, 周巳入³, 江凌勇^{3, *}, 代庆刚^{2, *}

1.上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面-头颈肿瘤科;
2.口腔第二门诊部;
3.口腔颅颌面科正颌-正畸中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室, 上海市口腔医学研究所,上海 200011

收稿日期:2019-01-16 修回日期:2019-03-15 出版日期:2019-05-20 发布日期:2019-06-21
通讯作者: 江凌勇,E-mail：jly117@sina.com;代庆刚,E-mail：daiqinggang@126.com。
作者简介:马旭辉（1987-）,女,博士,住院医师,E-mail：dr_mxh@126.com
基金资助:
国家自然科学基金（81570950,81870740,81800949）; SMC-晨星青年学者奖励计划优秀青年教师（B类计划）; 上海高校高峰高原学科建设项目; 上海交通大学医学院附属第九人民医院“精才培育项目”（JC201809）

Effects of puerarin on osteoclast differentiation of marrow-derived macrophages from mandible

MA Xu-hui¹, ZHANG Peng², YANG Yi-ling³, XU Hong-yuan³, GONG Xin-yi³, ZHOU Si-ru³, JIANG Ling-yong³, DAI Qing-gang²

1.Department of Oromaxillofacial Head and Neck Oncology;
2.The 2nd Dental Centre;
3.Department of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine;National Clinical Research Center for Oral Diseases;Shanghai Key Laboratory of Stomatology &Shanghai Research Institute of Stomatology. Shanghai 200011, China;

Received:2019-01-16 Revised:2019-03-15 Online:2019-05-20 Published:2019-06-21

摘要/Abstract

摘要： 目的: 分析不同浓度的葛根素对于下颌骨骨髓源巨噬细胞（Bone marrow-derived macrophages,BMMs）破骨分化的影响。方法: 体外分离培养4周龄雌性大鼠下颌骨BMMs。通过CCK-8分析1 nmol/L、10 nmol/L、100 nmol/L、1 μmol/L、10 μmol/L和100 μmol/L葛根素对BMMs细胞活性的影响。使用RANKL诱导BMMs破骨分化的同时加入不同浓度葛根素,采用抗酒石酸酸性磷酸酶（TRAP）酶化学染色及q-PCR分析破骨细胞形成及破骨相关基因的改变。采用SPSS16.0软件包对数据进行统计学分析。结果: 1 nmol/L、10 nmol/L、100 nmol/L、1 μmol/L、10 μmol/L葛根素组与对照组活细胞数无显著差异,100 μmol/L组活细胞数较对照组降低（P<0.05）。RANKL可诱导BMMs形成多核破骨细胞。10 nmol/L、100 nmol/L及1 μmol/L葛根素组破骨细胞数目及破骨细胞面积均较对照组降低（P<0.05）。q-PCR分析发现,10 nmol/L、100 nmol/L及1 μmol/L葛根素组NFATc1、C-fos、TRAP及CTSK mRNA表达量均较对照组下降（P<0.05）。结论: 葛根素可抑制下颌骨BMMs破骨分化及相关基因表达,为葛根素应用于下颌骨骨质疏松症的预防与治疗提供了依据。

关键词: 葛根素, 骨质疏松, 下颌骨骨髓源巨噬细胞, 破骨细胞

Abstract: PURPOSE: To study the effect of puerarin on osteoclast differentiation of marrow-derived macrophages (BMMs) from mandible. METHODS: BMMs were obtained from 4-week-old female SD rats. Cell viability of BMMs with different concentration of puerarin was determined by CCK-8. Tartrate resistant acid phosphatase staining and q-PCR were used to analyse the changes of osteoclast formation and expression of osteoclast gene of BMMs due to different concentration of puerarin. Statistical analysis was performed using SPSS 16.0 software package. RESULTS: Cell viability of BMMs with puerarin at 1 nmol/L, 10 nmol/L, 100 nmol/L, 1 μmol/L and 10 μmol/L showed no difference from the control group, and 100 μmol/L group displayed decreased cell viability (P<0.05). Osteoclast number and relative osteoclast surface reduced in BMMs with 10 nmol/L, 100 nmol/L and 1 μmol/L puerarin in comparison with the control group. Furthermore, the expression of NFATc1, C-fos, TRAP and CTSK deceased in BMMs with 10 nmol/L, 100 nmol/L, and 1 μmol/L puerarin. CONCLUSIONS: Puerarin could impede osteoclast differentiation of mandibular BMMs, which may be used to prevent and treat osteoporosis in alveolar and jaw bone.

Key words: Puerarin, Osteoporosis, Mandible bone marrow-derived macrophages, Osteoclast

中图分类号:

R739.8

马旭辉, 张鹏, 杨屹羚, 徐弘远, 龚心怡, 周巳入, 江凌勇, 代庆刚. 葛根素对大鼠下颌骨骨髓源巨噬细胞破骨细胞向分化的影响[J]. 中国口腔颌面外科杂志, 2019, 17(3): 193-197.

MA Xu-hui, ZHANG Peng, YANG Yi-ling, XU Hong-yuan, GONG Xin-yi, ZHOU Si-ru, JIANG Ling-yong, DAI Qing-gang. Effects of puerarin on osteoclast differentiation of marrow-derived macrophages from mandible[J]. China J Oral Maxillofac Surg, 2019, 17(3): 193-197.

参考文献

[1] 代庆刚,房兵,张鹏,等. 不同去势时间大鼠牙槽骨微结构变化的Micro-CT研究[J]. 上海口腔医学, 2014, 23(6): 641-645.
[2] Dai QG, Zhang P, Wu YQ, et al.Ovariectomy induces osteoporosis in the maxillary alveolar bone: an in vivo micro-CT and histomorphometric analysis in rats[J]. Oral Dis, 2014, 20(5): 514-520.
[3] Stewart S, Hanning R.Building Osteoporosis Prevention into Dental Practice[J]. J Can Dent Assoc, 2012, 78: c29.
[4] Guiglia R, Di Fede O, Lo RL, et al.Osteoporosis, jawbones and periodontal disease[J]. Med Oral Patol Oral Cir Bucal, 2012, 18: e93-99.
[5] Koduganti RR, Gorthi C, Reddy PV, et al.Osteoporosis: "A risk factor for periodontitis"[J]. J Indian Soc Periodontol, 2009, 13(2): 90-6.
[6] Rachner TD, Khosla S, Hofbauer LC.Osteoporosis: now and the future[J]. Lancet, 2011, 377(9773): 1276-1287.
[7] 刘建立. 应用雌激素防治绝经后骨质疏松症的问题[J]. 国际内分泌代谢杂志, 2006, 26(4): 230-232.
[8] Lin RC, Li TK.Effects of isoflavones on alcohol pharmacokinetics and alcohol-drinking behavior in rats[J]. Am J Clin Nutr, 1998, 68(6 Suppl): 1512S-1515S.
[9] Tan Y, Liu M, Wu B. Puerarin for acute ischaemic stroke[J]. Cochrane Database Syst Rev, 2008(1): CD004955.
[10] Wang Q, Wu T, Chen X, et al. Puerarin injection for unstable angina pectoris[J]. Cochrane Database Syst Rev, 2006(3): CD004196.
[11] Yang X, Yang Y, Zhou S, et al.Puerarin stimulates osteogenic differentiation and bone formation through the ERK1/2 and p38-MAPK signaling pathways[J]. Curr Mol Med, 2018, 17(7): 488-496.
[12] Dai Q, Zhou S, Zhang P, et al.Force-induced increased osteogenesis enables accelerated orthodontic tooth movement in ovariectomized rats[J]. Sci Rep, 2017, 7(1): 3906.
[13] Qiu ZK, Zhang GH, Zhong DS, et al.Puerarin ameliorated the behavioral deficits induced by chronic stress in rats[J]. Sci Rep, 2017, 7(1): 6266.
[14] 章明夏. 葛根素在绝经后骨质疏松桡骨远端骨折手术患者中的应用[J]. 按摩与康复医学(下旬刊), 2012, 03(5): 29-30.
[15] Yuan SY, Sheng T, Liu LQ, et al.Puerarin prevents bone loss in ovariectomized mice and inhibits osteoclast formation in vitro[J]. Chin J Nat Med, 2016, 14(4): 265-269.
[16] 李灵芝,刘启兵,姜孟臣,等. 葛根素对体外破骨细胞性骨吸收的影响[J]. 第三军医大学学报, 2004, 26(20): 1830-1833.
[17] 刘浩,李斌斌. 葛根素抑制1,25-二羟基维生素D3促破骨细胞骨吸收功能的最佳剂量[J]. 中国组织工程研究, 2012, 16(28): 5141-5144.
[18] 黄漫华,葛鸿庆,陈文治,等. 葛根素诱导破骨细胞自噬的机制研究[J]. 中国医药导报, 2015,12(15): 20-23.
[19] Zhou S, Yang Y, Ha N, et al.The specific morphological features of alveolar bone[J]. J Craniofac Surg, 2018, 29(5): 1216-1219.
[20] Liu H, Li W, Jia S, et al.Puerarin and zinc additively prevent mandibular bone loss through inhibiting osteoclastogenesis in ovariectomized rats[J]. Histol Histopathol, 2017, 32(8): 851-860.
[21] Dai Q, Han Y, Xie F, et al.A RANKL-based osteoclast culture assay of mouse bone marrow to investigate the role of mTORC1 in osteoclast formation[J]. J Vis Exp, 2018,2018(133).

葛根素对大鼠下颌骨骨髓源巨噬细胞破骨细胞向分化的影响

Effects of puerarin on osteoclast differentiation of marrow-derived macrophages from mandible

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 8

编辑推荐

Metrics

本文评价

[1]	金夏悦, 顾迎新. 骨质疏松对口腔种植修复影响的研究进展[J]. 中国口腔颌面外科杂志, 2023, 21(1): 81-86.
[2]	陈富民, 代庆刚, 邬春兰, 王跃平. 大豆异黄酮对颌骨骨髓源巨噬细胞破骨细胞向分化的影响[J]. 中国口腔颌面外科杂志, 2021, 19(3): 197-200.
[3]	李剑平, 刘培, 高学舸, 石燕萍. 雷洛昔芬对骨质疏松性颌骨骨折大鼠骨折愈合及OPG/RANKL/RANK系统的影响[J]. 中国口腔颌面外科杂志, 2021, 19(3): 213-216.
[4]	杨正祥, 李航, 李鲲. 雷洛昔芬对牙周炎合并系统性绝经后骨质疏松症小鼠模型局部牙槽骨破坏的影响[J]. 中国口腔颌面外科杂志, 2020, 18(5): 407-411.
[5]	李璐, 邱宇, 黄立, 蒋志豪, 王日辉, 高炳菊, 李军, 廖云阳, 林李嵩. 骨质疏松SD大鼠药物相关性颌骨骨坏死模型的建立[J]. 中国口腔颌面外科杂志, 2019, 17(5): 403-411.
[6]	魏立, 周琦, 江莉婷, 李宁, 高益鸣. 利塞膦酸钠对骨质疏松大鼠牙槽骨Bcl-2、BAX表达的影响[J]. 中国口腔颌面外科杂志, 2018, 16(5): 402-408.
[7]	杨锋, 孙玉华, 吕中静, 刘佃滨, 秦莹, 肖丽婷, 孙晋虎. 骨碎补总黄酮对骨质疏松症患者牙槽骨的影响[J]. 中国口腔颌面外科杂志, 2018, 16(1): 34-40.
[8]	马吴迪，夏伦果，周宇宁，徐袁瑾. COX-2与骨代谢关系的研究进展[J]. 中国口腔颌面外科杂志, 2016, 14(1): 83-88.