胰岛素、雌激素和培养时间对新生大鼠下颌骨成骨细胞增殖的影响  

Effects of insulin,estrogen and culture time on the proliferation of newborn rat mandibular osteoblasts

作　　者：鄂玲玲[1] 徐文焕[2] 乔朋艳[1] 邢鹤琳 王兴[1] 冯琳[1] 吕燕[1] 王东胜[1] 刘洪臣[1] E Ling-ling XU Wen-huan QIAO Peng-yan XING He-lin WANG Xing FENG Lin LV Yan WANG Dong-sheng LIU Hong-chen(Stomatologic institute, Chinese PLA General Hospital, Beijing 100853, China)

机构地区：[1]解放军总医院口腔医学研究所,副研究员北京100853 [2]解放军总医院医务部训练(研究生)处,助理员北京100853

出　　处：《中华老年口腔医学杂志》2017年第5期257-260,269,共5页Chinese Journal of Geriatric Dentistry

基　　金：国家自然科学基金(项目编号:81271180);解放军总医院临床科研扶持基金(项目编号:2016FC-CXYY-2007;2015FC-CXYY-1003)

摘　　要：目的:探讨胰岛素、雌激素和培养时间对体外培养的新生大鼠下颌骨成骨细胞(newborn rat mandibular osteoblasts,NRMOBs)增殖的影响。方法:取新生6-7天SD乳鼠下颌骨,原代培养下颌骨成骨细胞,选取生长状态良好的第4代细胞,在糖生理浓度(5.5mM)下与6ug/mL胰岛素(Insulin)和/或10^(-9) mol/L雌激素(E2)共同孵育,于1,3,5,7,9d采用MTT(Methylthiazol Tetrazolium,MTT)法检测胰岛素和雌激素对体外培养的NRMOBs增殖的影响。结果:所培养的NRMOBs阳性表达波形丝蛋白、骨钙素,角蛋白表达阴性,分泌碱性磷酸酶,形成钙结节。NRMOBs在5.5mM葡萄糖下培养,1d时,雌激素显著抑制细胞增殖;7d,9d时,胰岛素显著促进细胞增殖,5.5mM+Insulin组细胞增殖显著高于5.5m M+E2组;雌激素与胰岛素联合应用对细胞的增殖无影响。结论:在不同的培养时间,胰岛素或雌激素单独应用均影响NRMOBs增殖,胰岛素与雌激素联合应用能调节胰岛素或雌激素单独应用时对NRMOBs增殖的影响。Objective： To investigate the effect of insulin（In）, estrogen（E2） and culture time on the proliferation of newborn rat mandibular osteoblasts（NRMOBs）. Methods： The NRMOBs were cultured in vitro. The effect of insulin（6 ug/mL）and/or estrogen（10^（-9） mol/L） and/or culture time on NRMOBs was investigated. The NRMOBs proliferation was examined after 1、3、5、7、9 days of culture using Methylthiazol Tetrazolium（MTT）. Results： The NRMOBs positively expressed vimentin and osteocalcin, negatively expressed keratin, secreted alkaline phosphatase（ALP）, and produced calcium nodules.When NRMOBs were cultured in the media with 5.5mM glucose, estrogen significantly decreased the cells proliferation at 1 day of culture, insulin significantly increased the cells proliferation at 7, 9 day of culture. Compared to 5.5mM＋E2,5.5 mM＋Insulin significantly increased NRMOBs proliferation at 7, 9 day of culture. The combination of insulin and estrogen had no effect on the proliferation of NRMOBs at every time point. Conclusion： This study suggests that insulin or estrogen alone affects NRMOBs proliferation in different culture time. The combination of insulin and estrogen can adjust the effect of insulin or estrogen alone on NRMOBs proliferation.

关 键 词：成骨细胞 胰岛素 雌激素 下颌骨 大鼠 

分 类 号：R781[医药卫生—口腔医学]