Effects of zoledronate on CaMKⅡ δ and down-stream gene expressions during osteoclast differentiation
Wang Hui;Liu Juanjuan;Qi Mengchun;Dong Wei;Li Ren;Sun Hong;Teaching and Researching Section of Oral and Maxillofacial Surgery,College of Stomatology,North-China University of Science and Technology;Teaching and Researching Section of Pathology,College of Basic Medicine,North-China University of Science and Technology;
Objective To study the effect of zoledronate(ZOL)on Ca~(2+)/calmodulin-dependent kinaseⅡ δ(CaMKⅡδ)and down-stream gene expressions during osteoclast differentiation.Methods Mouse osteoclast precursors RAW264.7cells were divided into the control group and ZOL group.The cells in both groups were induced with 50μg/L receptor activator of nuclear factor kappa B ligand(RANKL)and were harvested on 5d,while the cells in ZOL group were also simultaneously treated with 1×10-6 mol/L ZOL for 2d.Five days later,the cells were harvested and examined osteoclastogenesis,as well as gene expressions of CaMKⅡδ,nuclear factor of activated T-cells cytoplasmic 1(NFATc1),tartrate-resistant acid phosphatase(TRAP)and cell-sarcoma receptor coactivator(c-Src).Results The number of TRAP positive multinuclear osteoclasts,number and size of dentin absorption lacunae and area in the ZOL group were(20.0±3.2),(18.0±4.2)and(6 335.3±1 043.2)μm2 respectively,which were significantly lower than(36.0±8.4),(37.2±5.0)and(11 636.2±3 661.1)μm2 in the control group and decreased by 44.4%,51.6% and 45.6% respectively(P0.01).ZOL also significantly inhibited the gene expressions of CaMKⅡ δ,NFATc1,TRAP and c-Src,and the mRNA levels of these genes were decreased by 44.1%,49.0%,53.8%and 49.6%respectively,the protein level were decreased by 43.5%,32.2%,45.5% and 48.0%respectively.The immunofluorescent cytochemistry detection results showed the fluorescence intensity of CaMKⅡδ,NFATc1,TRAP and c-Srcin in the ZOL group was significantly weakened when compared with the control group.Conclusion ZOL could significantly inhibit the osteoclast formation and bone absorption function,and down-regulates gene expressions of CaMKⅡ δ,NFATc1,TRAP and c-Src in osteoclast differentiation.
【Fund】： 国家自然科学基金资助项目(81270965);; 河北省教育厅重点项目(ZD2015005)
【CateGory Index】： R78
【CateGory Index】： R78