论文部分内容阅读
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 effects of insulin, estrogen and culture time on the proliferation of newborn rat mandibular osteoblasts (NRMOBs) cultured in vitro. Methods: Mandibular osteoblasts were primarily cultured in neonatal 6-7 day old SD rats. The fourth generation of cells were cultured in good condition and incubated with 6ug / mL Insulin in physiological concentration of glucose (5.5mM) And / or 10 ~ (-9) mol / L estrogen (E2) respectively. MTT assay was used to detect the proliferation of NRMOBs cultured in vitro by insulin and estrogen on MTT (Methylthiazol Tetrazolium) Impact. Results: The cultured NRMOBs positive expression of wave silk fibroin, osteocalcin, keratin expression was negative, the secretion of alkaline phosphatase, the formation of calcium nodules. NRMOBs were cultured in 5.5mM glucose, and estrogen significantly inhibited cell proliferation at 1d. Insulin significantly promoted cell proliferation at 7d and 9d, and significantly increased at 5.5mM + Insulin than 5.5m M + E2. Estrogen and insulin Combined application has no effect on cell proliferation. CONCLUSION: Insulin or estrogen alone affects the proliferation of NRMOBs at different culture times. The combination of insulin and estrogen can regulate the effect of insulin or estrogen on the proliferation of NRMOBs.