低渗作用对人成骨样细胞MG63功能的影响

Effects of Hypotonic Solution on the Proliferation,Alkaline Phosphatase Activity and [Ca^(2+)]i Concentration of Osteoblast-like Cells

目的本试验通过研究低渗透压的静牵张作用对成骨样细胞MG63的增殖能力、碱性磷酸酶活性以及[Ca2+]i的影响,探讨该应力形式下成骨样细胞MG63的力学响应特征。方法用不同渗透压的低渗透液,对成骨样细胞MG63分别进行2h、4h、8h、12h、24h和48h持续牵张作用后,用MTT法检测细胞增殖情况,用ALP试剂盒检测ALP活性的变化,用钙试剂盒检测[Ca2+]i含量波动情况。结果随着作用时间的延长,成骨样细胞MG63在277mmol/L和240mmol/L低渗透液的持续性膨胀作用下,其细胞增殖能力增强,[Ca2+]i、ALP活性缓慢增高;而细胞在163mmol/L低渗液作用下,细胞增殖受到抑制,8h时出现[Ca2+]i急剧升高,ALP活性明显高于其对照组。结论低渗膨胀对MG63的增殖分化、ALP活性以及Ca2+-ATPase都有一定的影响作用,且Ca2+内流与ALP活性之间存在一定的相关性。

Objective To investigate the effects of static-stretch from the hypotonic solution on the proliferation, alkaline phosphatase activity and [Ca^2＋]in the osteoblast-like cells. Methods Mechanical loading was introduced by swelling in the hypotonic solution. In vitro cultured MG63 were incubated under continuous swelling of 277, 240 and 163mOsm for 2h, 4h, 8h, 12h, 24h and 48h. The cell proliferation was detected by MTT assay. ALPase and [Ca^2＋] were determined by modified enzyme dynamic method and OCPC respectively. Results The cell proliferation, ALP activity and [Ca^2＋] increased slowly under continuous static-stretch of 277 and 240mOsm. The cell proliferation was inhibited under 163mOsm, with a sharp increase of [Ca^2＋] at 8h （11. 383±0. 111） and an increase of ALPase activity （0. 326±0. 002）. Conclusion The static-stretch induced from the hypotonic solution has an impact on the proliferation, differentiation, ALPase and Ca^2＋ -ATPase of the MG63. The [Ca^2＋] is correlated with the ALPase.