Pannexin1通道蛋白对微张力刺激下骨髓间质干细胞成骨分化的影响

被引：2

作者：

李盛龙 ^{[1
]}

王喆 ^{[2
]}

张晓晶 ^{[1
]}

汤欣 ^{[2
]}

机构：

[1] 沈阳, 中国医科大学肿瘤医院辽宁省肿瘤医院骨与软组织外科

[2] 大连医科大学附属第一医院创伤骨科

来源：

中华骨科杂志 | 2017年 / 37卷 / 01期

关键词：

间质干细胞; 细胞分化; 物理刺激;

D O I：

暂无

中图分类号：

R329.2 [人体细胞学];

学科分类号：

100107 [人体解剖与组织胚胎学（人体解剖学、组织与胚胎学）];

摘要：

目的探讨Pannexin1 (Px1)通道蛋白对微张力刺激下骨髓间质干细胞(mesenchymal stem cells, MSCs)成骨分化的影响。方法自Sprague Dawley大鼠(3周, 雌雄不限, 100～120 g)股骨和胫骨中提取全骨髓细胞进行原代培养。使用Px1通道蛋白抑制剂——甘珀酸/生胃酮(carbenoxolone , CBX)的安全剂量100 μmol/L(通过CCK8细胞活性检测试剂测得)进行实验。利用全骨髓培养法培养原代大鼠MSCs, 细胞传代至3～4代时, 获得纯度较好的MSCs。将MSCs随机分为三组:对照组(MSCs既不经CXB处理也不施加微张力刺激)、微张力刺激组(4 000 μstrain )和微张力刺激+CBX组。微张力刺激时间为15 min, CBX预处理时间分别为3 h、6 h、12 h和24 h。测定碱性磷酸酶活性、Ⅰ型胶原蛋白的表达、细胞内钙离子(Ca2+)的浓度, 以及Px1通道蛋白、磷酸化p38丝裂原活化蛋白激酶(p-p38 mitogen-activated protein kinase, p-p38MAPK)和p-ERK的表达活性。结果碱性磷酸酶(alkaline phosphatase , ALP)表达活性在微张力刺激组最高, 而使用CBX预处理后表达则不会升高。微张力刺激也可上调Ⅰ型胶原蛋白的表达, 增加Ca2+浓度, 上调Px1通道蛋白及p-p38MAPK的表达, 使用CBX预处理会降低以上效果。但微张力刺激下p-ERK的下调表达不受CBX预处理的影响。结论微张力刺激可促进MSCs成骨分化, 但使用CBX预处理会抑制微张力刺激的促进作用;其可能机制是CBX抑制了p-p38MAPK的活性。

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