Kamijo Minoru, Haraguchi Takayuki, Tonogi Morio, Yamane Gen-Yuki
Department of Oral Medicine, Oral and Maxillo-Facial Surgery, Tokyo Dental College, Chiba, Japan.
Biomed Res. 2006 Dec;27(6):289-95. doi: 10.2220/biomedres.27.289.
Connexin (Cx) 43-mediated gap-junctional intercellular communication (GJC) mainly regulates the osteoblastic differentiation, but much of the function of Cx43 on the differentiation of bone marrow cells is unclear. This study is aimed to clarify relationship between the differentiation of rat bone marrow cells and the function of Cx43. Bone marrow cells derived from four-week-old Wistar strain rats were grown in the presence and absence of 18-alpha-glycyrrhetinic acid (AGA, 100 muM) to inhibit Cx43-mediated GJC. Expression of Cx43 gene and protein, and the level of intracellular cyclic adenosine monophosphate (cAMP) were determined as the assessment of the function in Cx43-mediated GJC, and alkaline phosphatase (ALP) activity and mineralization were measured as the assessment of osteoblastic differentiation. The Cx43 gene expression was first observed at 2 days, but under the condition in which rat bone marrow cells were treated with AGA, there was no significant effect on the Cx43 gene expression. By administrating AGA to rat bone marrow cells, all parameters of maturation but the Cx43 gene expression significantly decreased. The results of this experiment suggest that Cx43-mediated GJC plays a critical role in rat bone marrow cells, progress toward maturation.
连接蛋白(Cx)43介导的间隙连接细胞间通讯(GJC)主要调节成骨细胞分化,但Cx43在骨髓细胞分化中的许多功能尚不清楚。本研究旨在阐明大鼠骨髓细胞分化与Cx43功能之间的关系。从四周龄Wistar品系大鼠中分离出的骨髓细胞,在存在和不存在18-α-甘草次酸(AGA,100 μM)的情况下培养,以抑制Cx43介导的GJC。测定Cx43基因和蛋白的表达以及细胞内环磷酸腺苷(cAMP)水平,作为对Cx43介导的GJC功能的评估,并测量碱性磷酸酶(ALP)活性和矿化程度,作为对成骨细胞分化的评估。Cx43基因表达在第2天首次观察到,但在大鼠骨髓细胞用AGA处理的条件下,对Cx43基因表达没有显著影响。通过向大鼠骨髓细胞施用AGA后,除Cx43基因表达外,所有成熟参数均显著降低。本实验结果表明,Cx43介导的GJC在大鼠骨髓细胞向成熟发展过程中起关键作用。