Cell-to-cell communication and expression of gap junctional proteins in human diabetic and nondiabetic skin fibroblasts -: Effects of basic fibroblast growth factor
Cell-to-cell communication and expression of gap junctional proteins in human diabetic and nondiabetic skin fibroblasts -: Effects of basic fibroblast growth factor
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DOI：
10.1385/endo:10:1:35
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发表时间：
1999-02-01
期刊：
影响因子：
3.7
通讯作者：
Grazul-Bilska, AT
中科院分区：
文献类型：
--
作者：
Abdullah, KM;Luthra, G;Grazul-Bilska, AT
伤口愈合涉及许多细胞类型的相互作用，并且部分由生长因子控制。缝隙连接介导的细胞间通讯被认为在组织器官生长发育的细胞代谢协调中起重要作用。碱性成纤维细胞生长因子（bFGF），已知是重要的伤口愈合，已被发现增加Cx43的表达和细胞间通讯的内皮细胞和心脏成纤维细胞。已经提出，增加的偶联对于这些细胞在伤口愈合和血管生成中的协调是必要的，并且bFGF的作用之一是调节细胞间通讯。本研究的目的是评估bFGF对体外细胞间隙连接通讯（CJIC）的影响，以及糖尿病和非糖尿病个体成纤维细胞中间隙连接蛋白（Cx）26、Cx 32和Cx 43的存在。将成纤维细胞系（n = 10）在含有或不含有bFGF（3 ng/mL）的无血清培养基中培养3d。用激光细胞仪检测细胞的GJIC率，用免疫组化和Western分析检测Cx 26、Cx 32和Cx43的存在。所有细胞类型通过接触依赖机制进行通信。糖尿病成纤维细胞的GJIC速率大于（p < 0.01）非糖尿病成纤维细胞（4.1 +/- 0.01 vs 3.3 +/-0.01%/min）。bFGF增加糖尿病（4.9 +/- 0.01 vs 4.1 +/- 0.01%）和非糖尿病（4.1 +/-0.01 vs 3.3 +/-0.01%）成纤维细胞的CJIC率（p < 0.01）。免疫组化检测到Cx 26位于细胞质中，Cx 32未检测到，Cx43存在于所有培养物的细胞边缘。免疫荧光染色图像分析显示bFGF增加糖尿病和非糖尿病成纤维细胞中Cx43的表达（p < 0.05）。Western免疫印迹分析显示，糖尿病和非糖尿病成纤维细胞在43-46 kD处的条带在体积上相似。因此，在成纤维细胞中涉及Cx43和GJIC的间隙连接似乎是bFGF的靶点。糖尿病个体的成纤维细胞似乎具有增加的细胞-细胞偶联速率，与降低的增殖速率相关。
Wound healing involves the interactions of many cell types, and is controlled in part by growth factors. Intercellular communication mediated by gap junctions is considered to play an important role in the coordination of cellular metabolism during the growth and development of tissues and organs. Basic fibroblast growth factor (bFGF), known to be important in wound healing, has been found to increase Cx43 expression and intercellular communication in endothelial cells and cardiac fibroblasts. It has been proposed that an increased coupling is necessary for the coordination of these cells in wound healing and angiogenesis, and that one of the actions of bFGF is to modulate intercellular communication. The aim of our study was to evaluate the effects of bFGF on gap junctional intercellular communication (CJIC) in vitro, and the presence of gap junctional proteins connexin (Cx) 26, Cx32, and Cx43 in fibroblasts of diabetic and nondiabetic individuals. Fibroblast cell lines (n = 10) were cultured for 3 d in serum-free media with or without bFGF (3 ng/mL). Cells were evaluated for the rate of GJIC by using laser cytometry, and for the presence of Cx26, Cx32, and Cx43 by immunohistochemical and Western analyses. All cell types communicated via contact-dependent mechanisms. The rate of GJIC was greater (p < 0.01) for diabetic than for nondiabetic fibroblasts (4.1 +/- 0.01 vs 3.3 +/- 0.01 %/min). bFGF increased (p < 0.01) the rate of CJIC for diabetic (4.9 +/- 0.01 vs 4.1 +/- 0.01%)and nondiabetic (4.1 +/- 0.01 vs 3.3 +/- 0.01 %)fibroblasts. Immunohistochemistry identified Cx26 in the cytoplasm, Cx32 was not detected, and Cx43 was present on the cellular borders in ail cultures. Image analysis of immunofluorescent staining demonstrated that bFGF increased (p < 0.05) Cx43 expression in diabetic and nondiabetic fibroblasts. Western immunoblot analysis revealed bands at 43-46 kD that were similar in volume for diabetic and nondiabetic fibroblasts. Thus, gap junctions involving Cx43 and GJIC among fibroblasts appear to be targets for bFGF. Fibroblasts of diabetic individuals appear to have an increased rate of cell-cell coupling, correlating with a decreased rate of proliferation.