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羊膜上皮细胞损伤后在纤维蛋白支架及细胞生长因子作用下修复状况的研究 被引量:2

Experimental study of repairing damaged human amniotic epithelial cells with formulated fibrin clot and cell growth factor
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摘要 目的探讨人羊膜上皮细胞(HAEC)损伤后能否在体外构建的纤维蛋白支架上修复,以及表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)和转化生长因子β1(TGF-β1)对HAEC增殖的影响。方法采用环钻钻切培养板上的HAEC建立定量损伤模型,用制备的纤维蛋白块覆盖环钻钻切范围,分别加入不同浓度的培养液进行培养。其中加入EGF为EGF组、加入bFGF为bFGF组、加入TGF-β1为TGF-β组,不加任何细胞生长因子作为对照组。显微镜下观察各组HAEC生长移行情况,5-溴脱氧尿嘧啶掺入法检测HAEC的增殖效应。结果(1)各组HAEC均能向环钻缺损处移行并向内生长,EGF组、bFGF组的HAEC移行速度快,细胞数量多,对照组次之,TGF-β1组最少。(2)EGF组不同浓度的EGF(1.0、5.0、10.0、20.0、40.0、80.0及160.0ng/ml)培养下,HAEC细胞增殖率分别为17.8%、28.0%、35.3%、51.6%、34.1%、34.2%及26.0%,EGF组10~80ng/ml的HAEC增殖效应均明显大于对照组(17.1%,P〈0.05)。(3)bFGF组不同浓度的bFGF(1.0、5.0、10.0、20.0.40.0、80.0及160.0ng/ml)培养下,HAEC细胞增殖率分别为18.0%、35.7%、43.0%、52.7%、67.4%、43.6%及30.5%。bFGF组5~80ng/ml的HAEC增殖效应均明显大于对照组(P〈0.05)。其中40ng/ml时的细胞增殖率最高(P〈0.05)。(4)TGF-β1组不同浓度的TGF-β1(0.1、0.2、0.4、0.8、1.6、3.2、6.4及12.8ng/ml)培养下,HAEC细胞增殖率分别为17.1%、15.1%、9.3%、6.2%、4.8%、3.6%、2.0%、1.2%。TGF-β1组0.8~12.8ng/ml的HAEC增殖效应明显小于对照组(P〈0.05)。结论通过纤维蛋白支架,HAEC能移行修复缺损部位。EGF、bFGF能促进HAEC的增殖,而TGF-β1则抑制HAEC的增殖。 Objective To investigate whether damaged human amniotic epithelial cells (HAEC) could be repaired on the matrix of formulated fibrin clot in vitro and the effects of epidermal growth factor (EGF) ,basic fibroblast growth factor(bFGF) and transforming growth factor β1 (TGF-β1)on the proliferation of HAEC. Methods Ring drill was used to drill the HAEC layer on culture sheets to make quantified models of damaged HAEC, on which the lacks were then covered with fibrin clot. Subsequently, EGF (EGF group), bFGF( bFGF group) and TGF-β1 (TGF-β1 group) of different concentration were added into the sheets respectively. After the predesigned culturing time, the growing and transiting conditions of HAEC were observed under inverted microscope after Giemsa stain. Also, the proliferating conditions of HAEC were detected by using 5-bromodeoxyuridine (BrdU). Results In all groups, HAEC could transit toward damaged area on fibrin clot and grow there. Higher transiting speed and larger cell numbers were observed in the EGF and bFGF groups followed by the control group, while the TGF-β1 group showed the relatively poorer results. Proliferating rates of HAEC were 17. 8% , 28.0%, 35.3%, 51.6% , 34. 1% , 34. 2% and 26. 0% respectively by EGF of different cultured concentration ( 1.0ng/ml, 5. 0ng/ml, 10. 0ng/ml, 20. 0ng/ml, 40. 0ng/ml, 80. 0ng/ml and 160. 0ng/ml) . Proliferating rates of HAEC were 18. 0%, 35. 7%, 43. 0% , 52. 7%, 67.4% , 43.6% and 30. 5% respectively by bFGF of different cultured concentration ( 1.0ng/ml, 5.0ng/ml, 10. 0ng/ml, 20. 0 ng/ml, 40. 0 ng/ml, 80. 0 ng/ml and 160. 0ng/ ml). Compared with the control group, EGF groups (EGF concentration ranging from 10ng/ml to 80ng/ml)and bFGF groups (bFGF concentration ranging from 5ng/ml to 80ng/ml ) showed better proliferating effects of HAEC (P 〈 0. 05 ), especially the 20ng/ml EGF group and 40ng/ml bFGF group had the best proliferating results among their own respective groups(P 〈0. 05). Proliferating rates of HAEC were 17. 1%, 15. 1%, 9. 3%, 6. 2%, 4. 8%, 3. 6%, 2. 0% and 1.2% respectively by TGF-β1 of different cultured concentration (0. 1 ng/ml, 0.2 ng/ml, 0.4 ng/ml, 0.8 ng/ml, 1.6 ng/ml, 3.2 ng/ml, 6.4 ng/ml and 12.8 ng/ml). Proliferating rates of HAEC in TGF-β1 groups (TGF-β1 concentration ranging from 0. 8ng/ml to 12. 8ng/ml) were significantly lower than that in the control group ( P 〈 0.05 ). Conclusions HAEC could transit and grow on the matrix of fibrin clot and repair the damaged area. EGF and bFGF could obviously stimulate HAEC proliferation ,while TGF-β1 might have the inhibitive effects.
出处 《中华妇产科杂志》 CAS CSCD 北大核心 2006年第1期12-15,共4页 Chinese Journal of Obstetrics and Gynecology
基金 重庆市自然科学基金资助项目(03-8006) 重庆市卫生局科研基金资助项目(03-2-100)
关键词 羊膜 上皮细胞 纤维蛋白 细胞分裂 细胞因子类 Amnion Epithelial cells Fibrin Cell division Cytokines
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参考文献7

  • 1漆洪波,李玮,孙江川,吴味辛,卞度宏.体外构建纤维蛋白支架封闭胎膜破口的研究[J].中国实用妇科与产科杂志,2005,21(6):377-378. 被引量:12
  • 2Mercer BM.Preterm premature rupture of the membranes.Obstet Gynecol,2003,101:178-193.
  • 3张斌,苏琦枫.胎膜早破的治疗[J].中华妇产科杂志,2002,37(1):53-54. 被引量:9
  • 4Yam HF,Pang CP,Fan DS,et al.Growth factor changes in ex vivo expansion of human limbal epithelial cells on human amniotic membrane.Cornea,2002,21:101-105.
  • 5王正国.王正国创伤外科学(第1版)[M].上海:上海科学技术出版社,2002.148-158.
  • 6Hoppenreijs VP,Pels E,Felten PC.Synergistic action of heparin and serum on basic fibroblast growth factor-modulated DNA synthesis and mitochondrial activity of cultured bovine corneal endothelial cells.Cornea,1996,15:386-396.
  • 7Moustakas A,Souchelnytskyi S,Heldin CH.Smad regulation in TGF-β signal transduction.J Cell Sci,2001,114:4359-4369.

二级参考文献25

  • 1赖仁胜,周溶.胎膜早期破裂的生物物理学机理探讨[J].中华妇产科杂志,1993,28(9):520-523. 被引量:14
  • 2Shubert PJ, Diss E, Iams JD. Etiology of preterm premature rupture of membranes. Obstet Gynecol Clin North Am, 1992,19:251-263.
  • 3Bou-Resli MN, Al-Zaid NS, Ibrahim ME. Full-term and prematurely ruptured fetal membranes: an ultrastructural study. Cell Tissue Res,1981,220:263-278.
  • 4Mac Deermott K. Amniotic membrane collagen content and type distribution in women with preterm premature rupture of membranes in pregnancy. Br J Obstet Gynecol,1998,105:369-370.
  • 5Kanayama N, Terao T, Kawashima Y, et al. Collagen types in normal and prematuely ruptured amniotic membranes. Am J Obstet Gynecol, 1985,153:899-903.
  • 6Hampson V, Liu D, Bilett E, et al. Amniotic membrane collagen content and type distribution in women with preterm premature rupture of the membranes in pregnancy. Br J Obstet Gynecol,1997, 104:1087-1091.
  • 7Guller S, Kong L, Wozniak R, et al. Reduction of extracellular matrix protein expression in humen amnion epithelial cells by glucocorticoids: a potential role in preterm rupture of the fetal membranes. J Clin Endocrinol Metab,1995, 80:2244-2250.
  • 8Vadillo OF, Arechevaleta F, Beltran MJ. Apoptosis and extracellular matrix degradation in chorion-amnio during labor and premature membrane rupture. Gynecol Obstet Mex,1998,66:202-207.
  • 9Lei H, Furth EE, Kalluri R, et al. A program of cell death and extracellular matrix degradation is activited in the amnion before the onset of labor. J Clin Invest, 1996,98:1971-1978.
  • 10Mercer BM, Arheart KL. Antimicrobial therapy in expectant management of preterm premature rupture of the membranes. Lancet,1995, 346:1271-1279.

共引文献19

同被引文献23

  • 1Romero R, Ghidini A, Bahado - Singh R. Premature rupture of membranes. In: Reece EA, ed. Medicine of the fetus and the mother[ J ]. Philadelphia (PA) : JB Lippincot,2004.
  • 2Elkhwad M, Pandev V, Stetzer B, et al. Fetal membranes from term vaginal deliveries have a zone of weakness exhibiting characteristics of apoptosis and remodeling [ J ]. J Soc Gynecol Investig, 2006,13 : 191 -- 195.
  • 3Wtanabe E , Smith DM, Sun J, et al. Effects of basic fibroblast growth factor on ang io gen esis in the in fracted porcine heart[ J]. Basic Res Cavadiol,2002,93:30 - 37.
  • 4Sagol S, Sagol O, Ozkal S, et al. Role of apotosis, Bcl - 2 and Bax protein expression in premature rupture of fetal membranes [ J]. J Reprod Med ,2005,47:809 - 815.
  • 5Kakishita K, Elwan MA, Nakao N, et al. Human amniotic epithelial cells produce dopamine and survive after implantation into the striatum of a rat model of Parkinson's disease: a potential source of donor for transplantation therapy. Exp Neurol 2000;165(1 ):27-34
  • 6Wei JP, Zhang TS, Kawa S, et al. Human amnion-isolated cells normalize blood glucose in streptozotocin-induced diabetic mice. Cell Transplant 2003;12(5):545-552
  • 7Takashima S, Ise H, Zhao P, et al. Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. Cell Struct Funct2004 ;29(3):73-84
  • 8In 't Anker PS, Scherjon SA, Kleijburg-van der Keur C, et al. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells 2004;22(7):1338-1345
  • 9Miao Z, Jin J, Chen L, et al. Isolation of mesenchymal stem cells from human placenta: comparison with human bone marrow mesenchymal stem cells. Cell Biol Int 2006;30(9):681-687
  • 10Zhao P, Ise H, Hongo M, et al. Human amniotic mesenchymal cells have some characteristics of cardiomyocytes, Transplantation 2005; 79(5):528-535

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