培养胚胎、小儿皮肤成纤维细胞的比较及tgf-β1对小儿成纤维细胞的干预研究

《培养胚胎、小儿皮肤成纤维细胞的比较及tgf-β1对小儿成纤维细胞的干预研究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、中图分类号：R321硕士学位论文培养胚胎、小儿皮肤成纤维细胞的比较及TGF-β1对小儿成纤维细胞的干预研究院（系）、所基础医学院研究生姓名何铭学科、专业病理与病理生理学导师姓名程基焱教授二Ο一二年四目录培养胚胎、小儿皮肤成纤维细胞的比较及TGF-β1对小儿成1纤维细胞的干预研究…………………………………………11.1中文摘要„„„„„„„„„„„„„„„„„„„„„11.2英文摘要„„„„„„„„„„„„„„„„„„„„„31.3前言„„„„„„„„„„„„„„„„„„„„„„„51.4.1第一部分材料与方法„„„„„„„„„„„„„„„„81.4.2

2、结果„„„„„„„„„„„„„„„„„„„„„„„151.4.3讨论„„„„„„„„„„„„„„„„„„„„„„„221.4.4结论„„„„„„„„„„„„„„„„„„„„„„„241.4.5第二部分材料与方法„„„„„„„„„„„„„„„„251.4.6结果„„„„„„„„„„„„„„„„„„„„„„„291.4.7讨论„„„„„„„„„„„„„„„„„„„„„„„371.4.8结论„„„„„„„„„„„„„„„„„„„„„„„401.5参考文献„„„„„„„„„„„„„„„„„„„„„411.6英汉缩略词对照表„„„„„„„„„„„„„„„„„46

3、2致谢„„„„„„„„„„„„„„„„„„„„„„„473无瘢痕愈合的信号转导机制研究进展（综述）„„„„„„„„„„„„„„„„„„„„„„„„„48胚胎、小儿皮肤成纤维细胞及TGF-β1干预后小儿皮肤成纤维细胞调节的Ⅰ、Ⅲ型胶原降解摘要目的：研究培养人胚胎、小儿皮肤成纤维细胞及TGF-β1干预后小儿皮肤成纤维细胞的盘状结构域受体1（DDR-1；discoidindomainreceptor）、基质金属蛋白酶13（MMP-13；matrixMetalloproteinases）、增殖细胞核抗原（PCNA；Proliferatingcellnuclea

4、rantigen）、Ⅰ型胶原（collagenⅠ；Col-Ⅰ）及Ⅲ型胶原(collagenⅢ；Col-Ⅲ)的表达及相互关系，从而探讨Ⅰ、Ⅲ型胶原的降解途径并观察培养胚胎与小儿成纤维细胞的形态特征。方法：1.原代培养小儿包皮成纤维细胞(humanpostnataldermalfibroblast；hPDF)和人胚胎皮肤成纤维细胞（humanfetaldermalfibroblast；hFDF）并传代；2.应用HE染色法观察hFDF与hPDF的形态结构特点；3.应用免疫组织化学S-P法检测hFDF与hPDF的DDR-1、MMP-13、PCNA（Prolif

5、eratingcellnuclearantigen）、Ⅰ型胶原（collagenⅠ；Col-Ⅰ）及Ⅲ型胶原(collagenⅢ；Col-Ⅲ)的表达；4.利用外源性转化生长因子β1（TGF-β1）（5ng/ml）干预hPDF,并设立TGF-β1未干预的对照组，分别于TGF-β1孵育8小时、24小时、48小时、96小时收获细胞；5.应用免疫组织化学S-P法观察不同时间点TGF-β1干预组与对照组hPDF的PCNA和DDR-1、MMP-13、Ⅰ型胶原及Ⅲ型胶原的表达；6.应用IX-71显微成像系统（OLYMPUS）在低倍（×100）视野下观察分析及照相，用I

6、mage-pro6.0图像分析系统测定各项指标的平均光密度值；7.所得数据以均数±标准差（±s）表示。采用spss19.0统计软件进行统计学分析，应用两独立样本t检验、秩和检验、重复测量方差分析、单因素方差分析、LSD检验、Pearson相关分析及多重线性回归分析法，P1＜0.05为具有统计学意义。结果：1.hFDF组DDR-1、MMP-13、Ⅰ、Ⅲ型胶原的平均光密度值皆高于hPDF组(P<0.05)，hFDF组与hPDF组的Ⅲ/Ⅰ胶原及PCNA的平均光密度值差异无统计学意义(P>0.05)。2.hPDFⅠ型胶原对DDR-1、Ⅲ型胶原、MMP-13的

7、线性回归方程显著成立ˆ(x：DDR-1；x：Ⅲ型胶原；x：MMP-13),Y=0.025-0.853x1+0.408x2+0.772x3123﹎其余各线性回归方程皆不成立。ˆ3.TGF-β1干预与对照组的两组细胞在不同ˆ时间点上ˆI型胶原、Ⅲ型胶原、DDR-1、MMP-13的平均光密度值不同（P<0.05），其中I型胶原、Ⅲ型胶原、MMP-13表达的基线上调（P<0.05）。ˆ4.DDR-1对Ⅰ、Ⅲ型胶原:Y=0.016+0.238x1-0.100x2(x1：I型胶原；x2：Ⅲ﹎ˆˆ型胶原)；Ⅰ型胶原对DDR-1、Ⅲ型胶原:Y=0.002+0.338x

8、1+1.084x2(x1：ˆ﹎Ⅲ型胶原；x2：DDR-1)；ˆⅢ型胶原对ˆⅠ型胶原、DDR-1