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目的观察１７β－雌二醇（Ｅ２）对成骨细胞胞内游离钙（［Ｃａ２＋］ｉ）、１，４，５－三磷酸肌醇（ＩＰ３）含量及钙调蛋白（ＣａＭ）含量的影响。方法以Ｆｌｕｏ－３／ＡＭ为钙荧光指示剂，采用激光共聚焦显微系统测定细胞内钙荧光值的改变，以反映［Ｃａ２＋］ｉ含量的变化。用阳离子交换法测定ＩＰ３含量。以测定钙依赖的磷酸二酯酶活性方法测定ＣａＭ含量。结果给予０．１、１．０ｎｍｏｌ／ＬＥ２后５０ｓ胞内钙荧光值分别增加４．７和６．１倍，持续８０～１６０ｓ；以１００ｎｍｏｌ／Ｌｔｈａｐｓｉｇａｒｇｉｎ预处理细胞，Ｅ２仅使胞内钙荧光值升高１．５倍。加入１．０ｎｍｏｌ／ＬＥ２，ＩＰ３含量在给药后２０～３０ｓ和６０～７０ｓ呈双峰升高。同样剂量的Ｅ２可使ＣａＭ含量增加８５．２％。他莫西芬不能阻断Ｅ２引起的胞内钙荧光值、ＩＰ３及ＣａＭ含量升高的作用，而磷脂酶Ｃ抑制剂使Ｅ２的作用部分或完全抑制。结论Ｅ２作用于胞膜引起胞外钙内流，并通过ＩＰ３导致［Ｃａ２＋］ｉ进一步增加，激活ＣａＭ从而调节细胞功能。
Objective To observe the effect of 17β-estradiol (E2) on intracellular free calcium ([Ca2 +] i), inositol 1,4,5-triphosphate (IP3) content and calmodulin (CaM) content in osteoblasts. Methods Fluo-3 / AM was used as an indicator of calcium fluorescence and the change of intracellular calcium fluorescence was measured by laser confocal microscopy to reflect the change of [Ca2 +] i content. Determination of IP3 by cation exchange method. CaM content was determined by measuring calcium-dependent phosphodiesterase activity. Results The fluorescence intensity of intracellular calcium in 50s was increased by 4.7 and 6.1 times, respectively, after treated with 0.1,1.0nmol / LE2 for 80 ~ 160s. Pretreatment of cells with 100nmol / Lthapsigargin, E2 only increased intracellular calcium fluorescence 1.5 times higher. With the addition of 1.0nmol / LE2, the IP3 content showed a doublet peak at 20 ~ 30s and 60 ~ 70s after administration. The same dose of E2 can increase the CaM content by 85.2%. Tamoxifen did not block E2-induced intracellular calcium fluorescence, increased IP3 and CaM levels, whereas phospholipase C inhibitors partially or completely suppressed the effects of E2. Conclusion E2 acts on the membrane of extracellular Ca2 + influx, and further increases [Ca2 +] i through IP3, activating CaM and regulating cell function.