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| 骨质疏松大鼠骨髓巨噬细胞的体外培养研究 |
| Culture of osteoporotic rat bone marrow-derived macrophages in vitro |
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| DOI：10.3969/j.issn.1006-7108.2019.04.004 |
| 中文关键词: 骨髓巨噬细胞 骨质疏松 去卵巢 极化 |
| 英文关键词:bone marrow-derived macrophages osteoporosis ovariectomy polarization |
| 基金项目:国家自然科学基金（81870766）；福建省科技创新联合资金项目（2016Y9023）；福建省卫生计生委青年科研项目（2016-1-72） |
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| 中文摘要: |
| 目的 体外分离培养骨质疏松大鼠骨髓巨噬细胞（bone marrow-derived macrophages,BMMs）并探讨其生物学行为。方法 采用双侧卵巢切除术建立大鼠绝经后骨质疏松症（postmenopausal osteoporosis ,PMO）动物模型并分离培养骨质疏松大鼠BMMs。通过倒置显微镜和瑞氏-姬姆萨染色对细胞进行形态学观察；通过CCK-8法检测骨质疏松大鼠BMMs的增殖能力；运用免疫荧光染色及荧光定量PCR法分析骨质疏松状态下大鼠BMMs的极化及炎症相关因子的表达情况。结果 成功建立大鼠PMO模型并分离培养骨质疏松大鼠BMMs。骨质疏松大鼠BMMs主要呈多角形及煎蛋样。与假手术组相比，BMMs的增殖能力较高。免疫荧光染色可见骨质疏松大鼠BMMs高表达CCR7，低表达MRC1。荧光定量PCR检测结果提示，骨质疏松大鼠BMMs促炎因子TNF-α及IL-6的表达较高，而抗炎因子IL-10及TGF-β的表达相对较低。结论 去卵巢可影响大鼠BMMs的增殖能力、细胞表面极化标志物以及炎症相关因子的表达水平。 |
| 英文摘要: |
| Objective To isolate and culture osteoporotic rat bone marrow-derived macrophages（BMMs）in vitro and explore their biological behaviors. Methods The postmenopausal osteoporosis（PMO）model of SD rats was established by bilateral ovariectomy. BMMs from osteoporotic rat were isolated and cultured in vitro. Inverted microscope and Wright Giemsa staining were employed for morphological observation of BMMs. The proliferation of BMMs from osteoporotic rat were detected by CCK-8 assay. The polarization and inflammation-related factors of BMMs in the PMO state were evaluated by immunofluorescence staining and real-time PCR. Results PMO model was established successfully. BMMs were mainly polygonal and omelette-like in osteoporotic rat. Comparing to sham group, the proliferation of osteoporotic rat BMMs significantly elevated. Immunofluorescence staining showed that BMMs of OVX group had higher expression of CCR7 and lower expression of MRC1. Real-time PCR demonstrated that the expression of pro-inflammatory factors of TNF-α and IL-6 increased in osteoporotic rat BMMs, while the expression of anti-inflammatory factors of IL-10 and TGF-β decreased. Conclusion Ovariectomy affects the proliferation, polarization and inflammation-related factors expression in rat BMMs. |
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