Effect of Advanced Oxidation Protein Products (AOPPs) and aging on the osteoclast differentiation of Myeloid-Derived Suppressor Cells (MDSCs) and its preliminary mechanism
Effect of Advanced Oxidation Protein Products (AOPPs) and aging on the osteoclast differentiation of Myeloid-Derived Suppressor Cells (MDSCs) and its preliminary mechanism
复制标题
高级氧化蛋白产物（AOPPs）和衰老对骨髓源性抑制细胞（MDSCs）破骨细胞分化的影响及其初步机制
DOI：
10.1016/j.bbrc.2022.10.066
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发表时间：
2022
影响因子：
3.1
通讯作者：
Siyuan Zhu
中科院分区：
文献类型：
--
作者：
Lin Zou;Wangsheng Jiang;Zihan Wang;Jianting Chen;Siyuan Zhu
为验证不同月龄小鼠骨髓间充质干细胞向破骨细胞分化的能力，探讨AOPPs对骨髓间充质干细胞向破骨细胞分化的影响。诱导破骨细胞分化后，这些细胞进行抗酒石酸酸性磷酸酶（TRAP）和F-肌动蛋白。将老年小鼠骨髓来源的MDSC注射到青年小鼠的胫骨髓腔内。一周后，对骨髓进行组织学检查、TRAP和细胞计数。从老年小鼠骨髓中分选MDSC用于诱导破骨细胞分化，用活性氧（ROS）清除剂、诱导型一氧化氮合酶（iNOS）抑制剂和一氧化氮（NO）清除剂干预，然后进行TRAP。给8周龄C57小鼠注射相同浓度的AOPP或小鼠血清白蛋白（MSA）。四周后，从骨髓中分选MDSC，并进行破骨细胞分化诱导，随后进行IHC染色和TRAP。提取8周龄C57小鼠骨髓间充质干细胞，用不同浓度的AOPPs体外诱导破骨细胞分化，然后进行TRAP训练，发现老年小鼠骨髓间充质干细胞数量明显高于青年小鼠。老年小鼠骨髓来源的MDSCs分化为多核TRAP+破骨细胞，其细胞数量和形态与中青年小鼠有显著差异。骨髓间充质干细胞分化的破骨细胞形成的肌动蛋白环在整个视野内密集分布，其密度明显高于中青年小鼠。老年小鼠骨髓间充质干细胞注射后，青年小鼠胫骨髓腔内TRAP+破骨细胞数量明显增加。NO抑制剂可显著抑制老年小鼠骨髓MDSCs向破骨细胞分化的能力，AOPPs体内处理可显著提高骨髓MDSCs的比例，最高可达55.2%。8周龄小鼠注射AOPPs诱导MDSCs分化为破骨细胞后，CD 11b+和Gr 1+细胞比例显著高于对照组和MSA组，但与15月龄小鼠差异不显著。不同浓度AOPPs体外诱导MDSCs无破骨细胞分化的迹象，MDSCs可直接向破骨细胞分化，老年小鼠骨髓MDSCs的分化能力较强;在体内AOPPs可以诱导MDSCs向破骨细胞分化，但在体外AOPPs不能诱导MDSCs向破骨细胞分化，提示体内可能存在其他因子与AOPP相互作用诱导MDSC向破骨细胞分化。
To verify the osteoclast differentiation ability of MDSCs from mice of different ages and explore the effect of AOPPs on the osteoclast differentiation of bone marrow MDSCs.Bone marrow cells from C57BL/6 (a.k.a C57) mice of different ages were subjected to flow cytometry, and CD11b+Ly6C+Ly6G+MDSCs were sorted out. After induction of osteoclast differentiation, these cells were subjected to tartrate-resistant acid phosphatase (TRAP) and F-actin. MDSCs from bone marrows of old mice were injected into the tibial medullary cavity of young mice. One week later, the bone marrows were subjected to histological examination, TRAP, and cell count. MDSCs from bone marrows of old mice were sorted for induction of osteoclast differentiation, intervened with reactive oxygen species (ROS) scavenger, inducible nitric oxide synthase (iNOS) inhibitor, and nitric oxide (NO) scavenger, and then subjected to TRAP. 8-weeks-old C57 mice were injected with the same concentrations of either AOPPs or mouse serum albumin (MSA). Four weeks later, MDSCs from bone marrows were sorted and subjected to induction of osteoclast differentiation, followed by IHC staining and TRAP. MDSCs of 8-weeks-old C57 mice were extracted and subjected toin vitroinduction of osteoclast differentiation with different concentrations of AOPPs, followed by TRAP training.The number of MDSCs in the bone marrows of old mice was significantly higher than that in young mice. MDSCs from bone marrows of old mice differentiated into large multinucleated TRAP+osteoclasts, which were significantly different from those in the middle-aged and young mice in terms of cell quantity and morphology. The actin rings formed in the differentiated osteoclasts from MDSCs of bone marrows were densely distributed in the whole field of view, which were significantly denser than those in the middle-aged and young mice. After injection of MDSCs of old mice, the number of TRAP+osteoclasts in the tibial medullary cavity of young mice was significantly increased. NO inhibitor can significantly inhibit the osteoclast differentiation capacity of MDSCs from bone marrows of old mice.In vivotreatment with AOPPs significantly increased the proportion of MDSCs in the bone marrow, which is up to 55.2%. After injection of AOPPs in 8-week-old mice and induction of osteoclast differentiation from the MDSCs, the ratios of CD11b+and Gr1+cells were significantly higher than that in the control and MSA groups but was not significantly different from that in the 15-month-old mice. Uponin vitrotreatment with different concentrations of AOPPs, the MDSCs did not show any sign of osteoclast differentiation.MDSCs can directly undergo osteoclast differentiation, the capacity of which is stronger in MDSCs of bone marrows of old mice; the NO pathway is a potential mechanism underlying this phenomenon.In vivobut notin vitroAOPPs treatment can induce osteoclast differentiation of MDSCs, indicating there might be other factors in the body that can interact with AOPPs to induce osteoclast differentiation of MDSCs.