Effect of high concentration of glucose on the production of TGF-?_1 by coculturing endothelial cells and smooth muscle cells / 第二军医大学学报

ABSTRACT

Objective: To study how to activate transforming growth factor ? 1 (TGF ? 1) by coculturing bovine cerebral microvascular endothelial cells (BCMEC) and bovine cerebral microvascular smooth muscle cells (BCMSMC) in vitro , and to observe the effect of high concentration of glucose on active TGF ? 1 (aTGF ? 1) production by the cocultured cells. Methods: BCMEC and BCMSMC were cocultured, and compared with BCMEC or BCMSMC homotypically cultured. 3H TdR assays were used to measure the bioactivity of aTGF ? 1 in the conditioned media. Dot blots assays were used to evaluate the TGF ? 1 mRNA levels of the cultured cells. Then, the cocultured cells were cultured with 5.5 (normal level), 15 and 25(high level) mmol/L glucose for 24 h. The activity of aTGF ? 1 in the media was measured by the same way. Results: aTGF ? 1 was produced in the media of cocultured cells, but not in the media of homotypically cultured cells. Activity of aTGF ? 1 in the media with high concentration of glucose was significantly higher than that in the media with normal concentration of glucose. The dot blots assays implicated that both BCMEC and BCMSMC had TGF ? 1 mRNA expression. But there was no significant change after coculturing. Conclusion: TGF ? 1 can be activated by contacted coculture of BCMEC and BCMSMC in vitro , but the coculturing does not increase the TGF ? 1 mRNA expression. Cultured with high concentration of glucose can increase the aTGF ? 1 content in the media of the cocultured cells. [