抑制Kv1.3表达能够降低人牙周膜成纤维细胞增殖和分化成骨能力  

Inhibition expression of Kv1.3 could reduce human periodontal ligament fibroblasts proliferation and osteogenic differentiation capacity

作　　者：朱庆勇[1] 陈文君[1] 朱慧[2] 沈洋[1] 祝心威[3] 蒋勇[4] 

机构地区：[1]中国人民解放军第105医院口腔科,合肥230031 [2]中国人民解放军第105医院检验科,合肥230031 [3]安徽医科大学口腔医学院,安徽医科大学附属口腔医院,安徽省口腔疾病研究中心实验室,合肥230032 [4]安徽医科大学第四附属医院口腔科,合肥230022

出　　处：《安徽医科大学学报》2017年第6期819-823,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省科技计划项目(编号:12070403062)

摘　　要：目的探讨Kv1.3在人牙周膜成纤维细胞(h PDLFs)增殖和分化成骨能力中的作用。方法通过体外分离并培养健康人和牙周炎患者牙周膜成纤维细胞,应用细胞免疫荧光法、Q-PCR和Western blot检测Kv1.3在牙周炎h PDLFs中表达变化;并通过si RNA沉默抑制Kv1.3表达,使用MTT和流式细胞仪观察对h PDLFs的增殖和分化成骨能力的作用。结果细胞免疫荧光结果显示与健康组比较,牙周炎组h PDLFs中Kv1.3显著低表达(P<0.05);Western blot结果显示Kv1.3-si RNA能够明显抑制细胞周期蛋白D1(Cyclin D1)和C-myc蛋白表达(P<0.05);流式细胞仪检测显示Kv1.3-si RNA能够显著抑制细胞G2/M期;同时,Western blot结果显示Kv1.3-si RNA明显降低碱性磷脂酶(ALP)和骨钙素(OCN)蛋白表达。结论抑制Kv1.3表达能够明显抑制h PDLFs的增殖和分化成骨能力。Objective To investigate the value of Kv1. 3 on proliferation and osteogenic differentiation capacity in human periodontal ligament fibroblasts （hPDLFs）. Methods hPDLFs were isolated and cultured in vitro from nor- mal and periodomtitis, the differential expression of Kv1. 3 was evaluated by immunofluoreseence, Q-PCR and Western blot. With siRNA silencing Kv1. 3 expression, the proliferation and osteogenic differentiation was evalua- ted in hPDLFs by flow cytometry and MTT. Results Compared with normal controls, Kvl. 3 expression was signifi- cantly lower in periodontitis hPDLFs by immunofluorescence assay. Western blot results showed that Kv1. 3-siRNA could inhibit CyclinD1 and C-myc protein expression（ P 〈 0. 05 ） ; Flow cytometry results showed that Kvl. 3-siRNA could significantly inhibit cell G2/M phase; Western blot results showed that Kv1. 3-siRNA decreased expression of ALP and OCN. Conclusion The expression of Kv1. 3 silencing can inhibit the proliferation and differentiation of osteogenie in bPDLFs significantly.

关 键 词：Kv1.3 人牙周膜成纤维细胞 增殖 分化 

分 类 号：R781.4[医药卫生—口腔医学]