摘要目的：观察人胚胎成纤维细胞（ FBs）对糖尿病足FBs生物学特性的影响，探讨其促进糖尿病溃疡愈合的机制。方法分离培养糖尿病创面FBs10例，分别与孕中期人胚胎皮肤FBs 3例（实验组）、正常人皮肤FBs 5例（对照组1）于transwell小室培养体系中间接共培养，并设立空白对照（对照组2）。绘制共培养体系中糖尿病足FBs生长曲线；观察共培养7 d后糖尿病足FBs形态；共培养至第4、7 d后，将各组糖尿病足FBs分离培养2 d，测量和对比上清液中羟脯氨酸和TGF-β1含量；检测共培养第3、5、7 d，β-半乳糖苷酶（ SA-β-gal ）染色阳性细胞百分率。胚胎FBs制作三维培养应用于8例糖尿病足溃疡，比较治疗前后创面愈合时间，做自身前后对照研究。结果实验组糖尿病足FBs比2个对照组增殖活跃，形态更接近于正常。经胚胎FBs处理4、7 d，糖尿病足FBs培养上清液中羟脯氨酸、TGF-β1明显增高，相对2个对照组差异均有统计学意义（羟脯氨酸4、7 d：与对照组1相比P＝0.023、P＝0.007；与对照组2相比P＝0.007、P＝0.003；TGF-β14、7 d：与对照组1相比P＝0.000、P＝0.000；与对照组2相比P＝0.000、P＝0.000）。实验组糖尿病足FBs的SA-β-gal染色阳性率在共培养第3、5及7 d无明显变化，2个对照组则明显增高。8例临床糖尿病足溃疡应用胚胎FBs三维培养治疗后创面均愈合。结论胚胎FBs能显著促进糖尿病足FBs增殖，延缓老化表型出现，同时上调羟脯氨酸、TGF-β1的分泌，可能与促进创面愈合机制有关。

abstractsObjective To observe the effects of human embryonic fibroblasts ( FBs ) on the biological characteristics of diabetic FBs in vitro , and to explore the possible therapeutic mechanism on diabetic ulcers . Methods Diabetic FBs were isolated and cultured with embryonic FBs (experimental group), adult FBs(control group 1)and alone(control group 2)in transwell chambers.Diabetic FBs growth curves in the coculture systems were described.The cell shape was observed by microscopy with HE staining .On the 4th and 7th day of cocul-ture, diabetic FBs were isolated to measure the expression levels of hydroxyproline and transforming growth factor -beta 1(TGF-β1)in the supernatant of monoculture .The cell senescense was displayed by β-galactosidase histo-chemical staining on the 3th, 5th and 7th day of coculture.Eight patients with diabetic foot ulcers who were can-didates for embryonic FBs constructs after failed traditional treatment were included in the study .The pre and post self-control study was conducted by observing the wound healing process before and after embryonic FBs treat -ment.Results Diabetic FBs in the experimental coculture system displayed more typical pattern and faster grow -ing rate.The level of hydroxyproline and TGF-β1 in the supernatant of diabetic FBs monoculture was significantly higher in the experimental group than in the control group after treated 4 or 7 days ( hydroxyproline:4 d, 7 d :compared with control group 1:P=0.023,P=0.007;compared with control group 2: P=0.007, P=0.003;TGF-β1 4 d, 7 d:compared with control group 1:P=0.000, P=0.000;compared with control group 2: P=0.000, P=0.000).The expression of SA-β-gal tended to be more apparent in control group 1 and 2 than in the experimental group on the 3rd,5th,and 7th day.All of the 8 cases with diabetic ulcers were cured by using em-bryonic FBs constructs .Conclusion Embryonic FBs can promote the proliferation of diabetic FBs , correct cell senescence and increase the production of hydroxyproline and TGF-β1,which maybe enhance the process of dia-betic wound healing .