巨噬细胞分化抗原-1通过脾酪氨酸激酶通路促进破骨细胞分化  被引量:5

Macrophage differentiation antigen- 1 promotes osteoclastogenesis via spleen tyrosine kinase pathway

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作  者:杨国曦 陈晓勇[1] 朱庆生[1] 杨重飞[1] 

机构地区:[1]第四军医大学附属西京医院骨一科,西安710032

出  处:《中华实验外科杂志》2018年第1期5-7,共3页Chinese Journal of Experimental Surgery

基  金:国家自然科学基金(81301541)

摘  要:目的探讨巨噬细胞分化抗原-1(Mac-1)分子在核因子-κB受体活化因子配体(RANKL)诱导下破骨细胞分化中的作用。方法用RANKL及巨噬细胞集落刺激因子(M-CSF)诱导C57BL/J6小鼠脾细胞,并使用抗CD11b及CD18抗体进行处理,1周后进行抗酒石酸酸性磷酸酶(TRAP)染色,测定细胞TRAP染色阳性率并进行统计学分析;用相同方法于载有骨片的96孔板上培养破骨细胞,利用抗CD11b、CD18抗体,干扰CD11b基因表达的慢病毒及其对照空载病毒转染,对骨片进行扫描电镜拍照并计算骨陷窝面积;提取各组细胞总核糖核酸(RNA)并进行反转录-聚合酶链反应(RT-PCR)及实时定量聚合酶链反应(Real-time PCR),测定各组蛋白质对应信使RNA(mRNA)表达水平。结果对照组、抗CD11b抗体组、抗CD18抗体组及双抗体组平均每孔TRAP阳性细胞数分别为(29.5±4.7)、(17.1±3.8)个(P=0.026)、(26.2±5.1)个(P=0.124)、(19.4±4.3)个(P=0.018),骨陷窝面积分别为(6.1±3.4)、(2.2±1.1)10-2 mm2/片(P=0.011)、(5.8±2.9)10-2 mm2/片(P=0.124)、(2.4±1.3)10-2 mm2/片(P=0.027),对照病毒组及病毒转染组骨陷窝面积分别为(5.6±3.2)、(1.3±0.9)10-2 mm2/片(P=0.006);相对于对照组和抗CD18抗体组,抗CD11b抗体处理的两组活化T细胞核因子-1(NFATc1)、脾酪氨酸激酶(Syk)和c-Fos mRNA表达下调。与对照病毒组比较,病毒转染组NFATc1、Syk和c-Fos mRNA表达下调。结论Mac-1分子通过其亚基CD11b上调Syk与c-Fos,促进NFATc1的表达,最终发挥促破骨细胞分化效应,而CD18亚基不具备促进作用。ObjectiveTo investigatethe effect of Mac-1 onosteoclast differention induced by RANKL and related signaling pathways.MethodsSpleen cells from 4-week-old C57BL/J6 mouse were treated with anti-CD11b, CD18 antibodies and inducedby(receptor activator for nuclear factor-κB ligand (RANKL), macrophage colony-stimulating factor (M-CSF). tartrate resistant acid phophatase (TRAP) staining was performed with TRAP kit; Spleen cells were cultured on bone silcesand treated with anti-CD11b, CD18 antibodies, lentivirus to silent CD11b gene and its control lentivirus, after induced by RANKL and M-CSF for 1week, bone resorption assay was performed; The mRNA levels of spleen tyrosine kinase (Syk), c-Fos and nuclear factor of activated T-cells, cytoplasmic-1 (NFATc1) were measuredusing reverse transcription-polymerase chain reaction (RT-PCR) and Real-time PCR.ResultsThe number of TRAP positive cells of control group, anti-CD11b antibody treatment group, anti-CD18 antibody treatment group and two antibodies treatment group were (29.5±4.7), (17.1±3.8) cells (P=0.026), (26.2±5.1) cells (P=0.124), (19.4±4.3) cells (P=0.018), the bone resorption area were (6.1±3.4), (2.2±1.1) 10-2 mm2/slice (P=0.011), (5.8±2.9) 10-2 mm2/slice (P=0.124), (2.4±1.3) 10-2 mm2/slice (P=0.027), bone resorption area of control lentivirus group and lentivirus group were (5.6±3.2), (1.3±0.9) 10-2 mm2/slice (P=0.006); Syk, c-Fos and NFATc1 mRNAlevels of groups treated with anti-CD11b antibodyor lentivirus weredownregulated.ConclusionMac-1 can promote osteoclast differentiation via Syk pathway activated by CD11b. CD18 does not have this effect.

关 键 词:骨质疏松 特异性抗体 慢病毒 

分 类 号:R580[医药卫生—内分泌]

 

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