天津医药

天津医药 ›› 2022, Vol. 50 ›› Issue (3): 236-240.doi: 10.11958/20212038

• 细胞与分子生物学 • 上一篇    下一篇

红景天苷抑制破骨细胞分化和极化的初步研究

易清清，梁鹏晨，孙苗苗，杨荣，梁冬雨，沙爽，常庆   

  1. 1上海健康医学院附属嘉定区中心医院中心实验室（邮编201800）；2上海中医药大学研究生院；3上海健康医学院附属嘉定区中心医院病理科；4上海健康医学院分子影像中心
  • 收稿日期:2021-09-03 修回日期:2021-11-19 出版日期:2022-03-15 发布日期:2022-03-15
  • 基金资助:
    上海市嘉定区卫健委重点项目（2020-ZD-03）；上海市嘉定区自然科学基金资助项目（JDKW-2020-0013）

A preliminary study of the inhibitory effect of salidroside on differentiation and polarization of osteoclasts

YI Qingqing, LIANG Pengchen, SUN Miaomiao, YANG Rong, LIANG Dongyu, SHA Shuang, CHANG Qing   

  1. 1 Clinical Research Center, Jiading District Central Hospital Affiliated to Shanghai University of Medicine & Health Sciences, Shanghai 201800, China; 2 Graduate School, Shanghai University of Traditional Chinese Medicine; 3 Department of Pathology, Jiading District Central Hospital Affiliated to Shanghai University of Medicine & Health Sciences; 4 Shanghai Key Laboratory for Molecular Imaging, Shanghai University of Medicine & Health Sciences
  • Received:2021-09-03 Revised:2021-11-19 Published:2022-03-15 Online:2022-03-15

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摘要： 目的　探究红景天苷抑制破骨细胞分化和极化的作用。方法　体外培养RAW264.7细胞，加入可溶性核因子κB受体活化因子配体（sRANKL）连续培养5 d诱导为破骨细胞。设置对照组（培养基不含红景天苷）和实验组（培养基中红景天苷剂量分别为15、30、60 mg/L），通过TRAP染色对比诱导分化结果，通过鬼笔环肽染色和茜素红染色观察破骨细胞F-Actin环形成和破骨细胞钙化情况，通过qPCR检测基质金属蛋白酶-9（MMP-9）、非受体酪氨酸激酶（c-Src）、组织蛋白酶K（CK）、整合素β3（Integrin β3） mRNA表达水平，Western blot检测MMP-9和c-Src蛋白表达水平。结果　与对照组相比，各实验组TRAP染色阳性细胞数显著减少，F-Actin环形成减少，而破骨细胞茜素红染色显著加深，钙化加重（均P＜0.05），且随着红景天苷剂量的升高，变化更加明显。与对照组相比，30和60 mg/L SAL组MMP-9、c-Src、CK mRNA表达水平均降低，同时各实验组MMP-9及c-Src蛋白表达水平显著降低。结论　红景天苷对破骨细胞的分化、极化和骨吸收有一定的抑制作用，其作用机制可能与下调MMP-9和c-Src表达有关。

关键词: 红景天属, 破骨细胞, 细胞分化, 红景天苷, 细胞极化, 骨吸收

Abstract: Objective　To investigate the inhibitory effect of salidroside (SAL) on osteoclast differentiation and polarization. Methods　RAW264.7 cells were cultured in vitro and induced into osteoclasts by adding solvable receptor activator of nuclear factor kappa B ligand (sRANKL) for 5 days. The control group (culture medium without SAL) and the experimental group (culture medium with SAL in dosage of 15, 30, 60 mg/L respectively) were set. The results of inducing differentiation were compared by TRAP staining. F-Actin ring formation and osteoclast calcification were compared by phalloidin staining and alizarin red staining. Real time fluorescent quantitative reverse transcription polymerase chain reaction (qPCR) was used to detect expression levels of MMP-9, c-Src, CK and Integrin β3. The expression levels of MMP-9 and c-Src proteins were detected by Western blot assay. Results　Compared with the control group, the number of TRAP positive cells, and F-Actin ring formation were significantly reduced in each experimental group. The calcification of osteoclasts and the alizarin red staining of osteoclasts were significantly deepened, indicating the the increased calcification (P＜0.05). Compared with the control group, the expression levels of MMP-9, CK and c-Src mRNA decreased in the 30 and 60 mg/L SAL groups, and the expression levels of MMP-9 and c-Src protein decreased significantly in each group. Conclusion　Salidroside can inhibit the differentiation, polarization and bone resorption of osteoclasts, and its mechanism may be related to the down-regulation of MMP-9 and c-Src expression.

Key words: rhodiola, osteoclasts, cell differentiation, salidroside, cell polarization, bone resorption