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锶离子对钛颗粒诱导破骨细胞活化的作用 被引量:5

Inhibitory effects of strontium on osteoclatogenesis induced by titanium particles
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摘要 目的 观察锶离子(Sr2+)对钛颗粒(Ti)诱导破骨细胞(OC)活化的影响。方法实验分为5组,即空白组、Ti组、核因子受体活化因子配体(RANKL)组、R+Ti组、Sr2+组。采用细胞增殖-毒性检测试剂盒(CCK-8)检测0.1 g/L Ti和不同浓度Sr2+(0.5、1.0、2.0、5.0、10.0 mol/L)处理小鼠单核/巨噬细胞株RAW264.7细胞24、48、72 h后增殖活性,以0 1 g/L Ti和/或70μg/L RANKL和/或不同浓度Sr2+诱导RAW264.7细胞,培养5 d后,用抗酒石酸酸性磷酸酶(TRAP)染色检测成熟OC数量,以逆转录-聚合酶链反应(RT-PCR)检测组织蛋白酶K(Cath-K)、TRAP、基质金属蛋白酶-9(MMP-9)和降钙素受体(CTR)mRNA含量,Westem blot法检测κB抑制蛋白α(IκBα)表达水平。结果 CCK-8结果,0.1 g/L Ti和不同浓度Sr2+(0.5、1.0、2.0、5 0、10.0 mmol/L)对RAW264.7细胞增殖能力无影响。TRAP染色,R、R+Ti组均可见大量的紫红色多核细胞,Ti组和Sr2+组多核细胞较少,Sr2+≥5 mmol/L时,TRAP阳性细胞数量[(323.33±43.84)、(146.67±33 99)个/孔],与R组[(453.33±33.99)个/孔]比较,差异有统计学意义(P〈0 05)。RT-PCR结果显示,Sr2+组Cath-K、TRAP、MMP-9和CTR mRNA的相对表达量分别为2.76±0.66、2.69±0.57、2.10±0.34和1.72±0.32,与R+Ti组比较(7.97±0.77、10.10±1.18、7.37±1.02和5.55±0.53),差异有统计学意义(P〈0.05)。Western blot结果显示Ti能抑制IκBα的表达,Sr2+加入后Ti的抑制效应不明显。结论 Sr2+能有效地抑制Ti诱导的OC活化。 Objective To observe the effect of strontium (Sr2+ ) on titanium (Ti) particles-in- duced osteoclastgogenesis from a murine macrophage cell line (RAW264. 7) in the presence of receptor ac- tivator of nuclear faetor-KB (NF-KB) ligand (RANKL). Methods The experiment involved 5 groups : black group, Ti group, R group, R + Ti group and Sr2 + group. The effect of Ti particles and (or) Sr2+ on RAW264.7 cell viability was measured by using the cell counting kit-8. Tartrate resistant acid phosphatase (TRAP) staining was used to analyze the osteoclast formation. Quantitative real-time reverse transcriptionpolymerase chain reaction (RT-PCR) was used to detect the mRNA levels of cathepsin K (Cath-K) calcitonin receptor (CTR) , matrix metalloproteinase-9 (MMP-9) and TRAP. The inhibitory KBc+ (IKBct) in NF-KB signaling pathway was determined by Western blotting. Results Sr2 + (0. 5, 1.0, 2.0, 5.0, and 10.0 mmol/L) did not affect the viability of RAW264. 7 cells cultured with Ti particles. The mature osteo- clasts were obtained from RAW264. 7 cells cultured with RANKL and Ti particles for 5 days and detected by TRAP staining. The number of TRAP positive cells in 5 mmol/L Sr2+ was (323.33 ±43.84)/well, which was significantly decreased as compared with R group [ (453. 33 ±33.99)/well] (P〈 0. 05). RT-PCR demonstrated that mRNA levels of Cath-K, CTR, MMP-9 and TRAP were significantly greater in medium with RANKL and Ti particles than without RANKL and Ti particles. When 5 mmol/L Sr2 + was added to the culture system for 5 days, the mRNA levels of Cath-K, TRAP, MMP-9 and CTR respectively were 2. 76±0. 66, 2. 69 ±0. 57, 2. 10±0. 34 and 1.72±0. 32, in combination with RANKL and Ti particles (7. 97 ± 0. 77, 10. 10 ± 1.18, 7.37 ± 1.02 and 5.55 ± 0. 53 ) ( P 〈 0. 05 ). The results of Western blotting showed that the levels of IKBct were reduced at 15 min but increased again at 180 min after Ti particle treatment. Pretreatment with Sr2 + significantly increased the expression of IKBct in response to Ti particle. Conclusion This study provides evidence that Sr2+ can markedly inhibit Ti particle-induced oste-oclastogenesis by the down-regulation of NF-KB signaling pathway, and is a promisingly therapeutic candidate for the wear particle-induced osteolysis.
出处 《中华实验外科杂志》 CAS CSCD 北大核心 2014年第7期1399-1401,F0003,共4页 Chinese Journal of Experimental Surgery
基金 国家自然科学基金资助项目(81101399、81272018、81372018) 江苏省自然科学基金资助项目(BK2011303) 江苏省临床医学科技专项资助项目(BL2012004) 江苏省研究生培养创新工程资助项目(CXZZ13-0835)
关键词 破骨细胞 钛颗粒 核因子-ΚB Strontium Osteoclast Titanium particle Nuclear factor-KB
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