电离辐射对破骨细胞分化过程中RANK表达的影响及其致骨损伤的分子机制  

Effect of ionizing radiation on expression of RANK in differentiation process of osteoclasts and its molecular mechanism of bone injury

作　　者：周慧[1] 杨冰[2,3] 唐泉[2] 孙元明[2] 韩英[2] 樊飞跃[2] 刘晓冬[4] 贾立立[4] 

机构地区：[1]天津市临床药物关键技术重点实验室天津医科大学药学院,天津300070 [2]北京协和医学院&中国医学科学院放射医学研究所放射生物研究室,天津300192 [3]天津医科大学基础医学院细胞生物学系,天津300070 [4]吉林大学公共卫生学院卫生部放射生物学重点实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2013年第6期1127-1131,1312,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30970867);吉林大学基本科研业务费资助课题(201103082)

摘　　要：目的:研究电离辐射对破骨细胞分化过程中核因子κB受体活化因子(RANK)mRNA和蛋白表达的影响,探讨辐射导致骨损伤的分子机制。方法:采用50μg·L-1核因子κB受体活化因子配体(RANKL)诱导RAW264.7细胞分化为破骨细胞。RQW264.7细胞分为对照组(未处理)、RANKL处理组(50μg·L-1RANKL处理)、照射处理组(2Gyγ射线照射)、照射联合RANKL处理组(50μg·L-1 RANK处理+2Gyγ射线照射)。采用抗酒石酸磷酸酶(TRAP)染色法检测各组破骨细胞的分化状态;采用PCR法检测各组细胞中RANK mRNA的表达水平;采用Western blotting法检测各组细胞中RANK蛋白的表达水平。结果:RAW264.7细胞经过RANKL诱导7d后,TRAP染色呈现阳性,表明已经成功分化成为破骨细胞。与对照组比较,RANKL处理组和照射处理组破骨细胞前体细胞中RANK mRNA和蛋白的表达水平上调(P<0.05);与RANKL处理组比较,照射联合RANKL组破骨细胞前体细胞中RANK mRNA和蛋白表达水平降低(P<0.05)。结论:电离辐射可以促进破骨细胞前体细胞的增殖和成熟,增加其活性,但是对破骨细胞的增殖、成熟与活性却有一定的抑制作用。Objective To investigate the effect of ionizing radiation on the expressions of receptor activator for NF-κB （RANK） mRNA and protein in differentiation process of osteoclasts and to discuss the molecular mechanism of bone injury induced by ionizing radiation. Metho6s The osteoblasts were differentiated from RAW264.7 cells after treated by 50μg·L-1 receptor aetivatorfor NF-κB ligand （RANKL）. The RAW264.7 cells were divided into control group （normal RAW264.7 cells without treatment）, RANKL treatment group （RAW264.7 cells treated with 50μg·L-1 RANKL）, radiation treatment group （RAW264.7 cells exposed to 2 Gy γ-rays） and radiation combined with RANKL treatment group （RAW264.7 cells treated with both 2 Gy γ-rays and 50 μg·L-1 RANKL）. Tartrate resistant acid phosphatase （TRAP） staining method was used to assess the osteoclast differentiation status; the expression levels of RANK mRNA and protein of ostoclasts were detected by PCR method and Western blotting method, respectivly. Results After treated with RANKL for 7 d, the RAW264.7 cells showed TRAP positive result, which indicated the formation of osteoclasts. Compared with control group, the expression levels of RANK mRNA and protein of osteoclast precursors in RANKL treatment group and radiation treatment group were increased （P〈0.05）. Compared with RANKL treatment group, the expression levels of RANK mRNA and protein in radiation combined with RANKL treatment group were decreased （P〈0.05 ）. Conclusion Ionizing radiation may promote the proliferation and mature, and increase the activity of osteoclast precursors, but it can inhibit the proliferation, mature, and activity of osteoclasts.

关 键 词：电离辐射 RAW264 7细胞 破骨细胞 RANK 放射治疗 

分 类 号：R144.1[医药卫生—公共卫生与预防医学]