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为研究不同切应力下内皮细胞条件培养基对平滑肌细胞和胶原合成的影响 ,建立平行平板流动腔模型 ,模拟产生定常层流 ,采用3H -胸腺嘧啶核苷掺入法和胃蛋白酶消化法 ,测定细胞DNA和胶原合成量。实验发现 ,与单用 10 %牛血清DMEM培养基比较 ,静态培养的内皮细胞条件培养基促使平滑肌细胞DNA和胶原合成 ,3H-胸腺嘧啶核苷掺入量从 749± 5 3cpm/ 10 3 细胞上升至 12 0 2± 6 3cpm/ 10 3(P <0 .0 1) ,平滑肌细胞掺入 2 ,3- 3H -羟脯氨酸量从 30± 6cpm/ 10 3 细胞上升至 47± 4cpm/ 10 3 细胞 (P <0 .0 1)。内皮细胞在 10和 2 5dyn/cm2 切应力剪切 18h后 ,内皮细胞条件培养基对平滑肌细胞DNA和胶原合成促进效应分别下降了 10 .41%± 5 .6 6 %、2 3.97%± 6 .2 3 %和 45 .71%± 2 .93%、6 4.5 3%± 2 .42 %。由此提示内皮细胞所受血流切应力减小后 ,其条件培养基能促使平滑肌细胞的增殖和胶原合成。
In order to study the effect of endothelial cell conditioned media on the synthesis of smooth muscle cells and collagen under different shear stress, a parallel plate flow cavity model was established to simulate the steady laminar flow. 3H - thymidine incorporation and pepsin digestion were used to determine Cell DNA and collagen synthesis. The results showed that compared with DMEM medium supplemented with 10% bovine serum alone, static culture of endothelial conditioned medium promoted the synthesis of DNA and collagen in smooth muscle cells, and the incorporation of 3H-thymidine increased from 749 ± 5 3 cpm / 10 3 cells To 122 ± 6 3 cpm / 103 (P <0.01), the amount of 3H-hydroxyproline incorporated into smooth muscle cells increased from 30 ± 6 cpm / 103 cells to 47 ± 4 cpm / 103 Cells (P <0.01). Endothelial cell conditioned media decreased the DNA and collagen synthesis-promoting effects of smooth muscle cells by 10.41 ± 5.66% and 2.97% ± 6, respectively, at 10 and 25dyn / cm2 for 18h. 2 3% and 45 .71% ± 2 .93%, 6 4.5 3% ± 2.42%. This suggests that the shear stress on endothelial cells is reduced, the conditioned medium can promote smooth muscle cell proliferation and collagen synthesis.