摘要
目的探讨晚期氧化蛋白终产物(AOPPs)对破骨细胞分化的影响。方法体外分离培养大鼠骨髓单核细胞,分为空白对照组、阴性对照组(RSA)、AOPPs组、阳性对照组(RANKL)、还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶抑制剂组(AOPPs+夹竹桃麻素),干预6 d后,通过抗酒石酸酸性磷酸酶(TRAP)染色观察TRAP阳性多核细胞,鬼笔环肽荧光染色观察F肌动蛋白(F-actin)环,甲苯胺蓝染色观察骨磨片的骨吸收陷窝。此外,AOPPs刺激2 h后,通过DCFH-DA荧光探针法检测细胞内活性氧(ROS)生成。结果 AOPPs及阳性对照组RANKL均可诱导TRAP阳性多核细胞、F-actin环及骨吸收陷窝形成,并且AOPPs刺激后细胞ROS生成明显增多。AOPPs所致以上效应均能被NADPH氧化酶抑制剂夹竹桃麻素阻断。结论 AOPPs可诱导大鼠骨髓单核细胞向破骨细胞分化。
Objective To evaluate the effect of advanced oxidation protein products （AOPPs） on osteoclast differentiation in vitro. Methods Rat bone marrow mononuclear ceils were isolated and cultured in vitro, and divided into the blank control, negative control （ RSA）, positive control（ RANKL）, AOPPs, NADPH oxidase inhibitor groups （ AOPPs＋ Apocynin）. After treatment for 6 days, the TRAP- positive multinuclear ceils were observed by using tartrate-resistant acid phosphatase （TRAP） staining, the F-actin ring by phalloidine fluo- rescence staining, the bone resorption pits on bone slices by toluidine blue. Furthermore, intracellular reactive oxygen species （ROS） gener-ation were detected by DCFH-DA fluorescent probe after AOPPs stimulation for 2 hours. Results The TRAP-positive muhinuclear cells, F- aetin ring and bone resorption pits on bone slices were observed in AOPPs and RANKL positive control groups. Meanwhile, AOPPs stimulation increased ROS generation. However, the above effects of AOPPs challenge were abolished by the apocynin, a NADPH oxidase inhibitor. Conclusions AOPPs could induce the osteoclast differentiation of rat bone marrow mononuclear cells in vitro, which provides a new sight in understanding the pathogenic basis of osteoporosis.
作者
庄景燊
朱思远
黄玉圣
徐平
陈建庭
钟招明
ZHUANG Jing-Shen ZHU SiYuan HUANG Yu-Sheng et al(Department of Orthopedic and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong, China)
出处
《中国老年学杂志》
CAS
北大核心
2017年第3期521-525,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金项目(No.81000785
81472135)
关键词
晚期氧化蛋白终产物
骨髓单核细胞
破骨细胞
分化
Advanced oxidation protein products
Bone marrow mononuclear Cells
Osteocalst
Differentiation
