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目的建立稳定表达人血小板衍生生长因子－ＢＢ（ｐｌａｔｅｌｅｔ－ｄｅｒｉｖｅｄｇｒｏｗｔｈｆａｃｔｏｒ－ＢＢ，ＰＤＧＦ－ＢＢ）的细胞株，并考察该重组蛋白与损伤修复有关的生物学作用。方法将含ＰＤＧＦ－Ｂ基因的真核表达载体ｐｃＤＮＡ３转染小鼠成纤维细胞ＮＩＨ３Ｔ３中，在Ｇ４１８选择压力下得到抗性克隆，收集扩增的抗性克隆培养液并制备细胞膜上ＰＤＧＦ－ＢＢ蛋白质，３Ｈ－ＴｄＲ掺入法表达产物的生物学活性，并以３Ｈ－Ｐｒｏ掺入测胶原合成率。结果免疫荧光染色法证实抗性细胞中ＰＤＧＦ－ＢＢ的表达，膜上ＰＤＧＦ－ＢＢ显著促进ＮＩＨ３Ｔ３细胞ＤＮＡ合成，活性可达约３１．２Ｕ／１０６细胞，而培养液的致丝裂活性低于可检测水平。抗性细胞的胶原合成率显著增高。结论ＰＤＧＦ－ＢＢ的真核表达系统已得以成功构建，该重组蛋白可促进成纤维细胞增生和胶原合成，提示其在损伤修复中有潜在应用价值
Objective To establish a cell line stably expressing platelet-derived growth factor-BB (PDGF-BB) and investigate the biological role of the recombinant protein in wound healing. Methods Eukaryotic expression vector pcDNA3 containing PDGF-B gene was transfected into NIH3T3 mouse fibroblasts. The resistant clones were obtained under the selective pressure of G418. The amplified resistant clonal medium was collected and the PDGF-BB protein The 3H-TdR incorporation method was used to express the biological activity of the product. 3H-Pro incorporation was used to measure the collagen synthesis rate. Results The expression of PDGF-BB in the resistant cells was confirmed by immunofluorescence staining. The PDGF-BB on the membrane significantly promoted the DNA synthesis of NIH3T3 cells with the activity up to about 31.2U / 106 cells, while the culture broth had less mitogenic activity than the Test level. The rate of collagen synthesis in resistant cells was significantly higher. Conclusion The eukaryotic expression system of PDGF-BB has been successfully constructed. The recombinant protein can promote the fibroblast proliferation and collagen synthesis, suggesting that it has potential value in the injury repair