摘要
目的 :探讨体外混合培养的牛脑微血管内皮细胞 (BCMEC)和平滑肌细胞 (BCMSMC)产生转化生长因子 β1 (a TGF-β1 )的条件 ,观察高浓度葡萄糖对混合培养细胞产生 a TGF- β1 的影响。 方法 :将 BCMEC与 BCMSMC在体外作混合接触培养 ,与单独培养的 BCMEC或 BCMSMC作对照。用 3H - Td R掺入法检测培养上清中 TGF-β1 的生物学活性。用点杂交法检测TGF-β1 m RNA表达水平的变化。分别用含 5 .5、15、2 5 mm ol/ L D-葡萄糖的培养液将 BCMEC和 BCMSMC混合培养 2 4h,检测其上清中 TGF- β1 的生物学活性。 结果 :混合培养的细胞上清中有 a TGF- β1 产生 ,而单独培养的细胞上清中均无。高糖培养的 BCMEC和 BCMSMC上清中 TGF- β1 活性较正常水平葡萄糖培养的显著增高。点杂交结果显示 ,单独培养的 BCMEC或 BCMSMC均有 TGF-β1 m RNA表达 ,混合培养后 m RNA水平无明显改变。 结论 :体外混合接触培养的 BCMEC和BCMSMC可以激活 TGF-β1 ,但并未增加 TGF-β1 的 m RNA表达。高糖可使混合培养细胞上清中 a TGF-β1 含量增加。
Objective: To study how to activate transforming growth factor β 1 (TGF β 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 β 1 (aTGF β 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 β 1 in the conditioned media. Dot blots assays were used to evaluate the TGF β 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 β 1 in the media was measured by the same way. Results: aTGF β 1 was produced in the media of cocultured cells, but not in the media of homotypically cultured cells. Activity of aTGF β 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 β 1 mRNA expression. But there was no significant change after coculturing. Conclusion: TGF β 1 can be activated by contacted coculture of BCMEC and BCMSMC in vitro , but the coculturing does not increase the TGF β 1 mRNA expression. Cultured with high concentration of glucose can increase the aTGF β 1 content in the media of the cocultured cells. [
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2001年第3期278-280,共3页
Academic Journal of Second Military Medical University
