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目的 研究第 2代人发角蛋白 (humanhairkeratinⅡ ,HHK -Ⅱ )人工腱植入后不同时期转化生长因子 - β1,2 ,3(TGF - β1,2 ,3)mRNA的表达及泛素 (ubiquitin)的分布。 方法 选用4只新西兰兔制作动物模型 ,用原位杂交 (insituhybridization ,ISH)的方法研究HHK -Ⅱ人工腱植入后不同时期TGF - β1,2 ,3mRNA的表达 ;用免疫组化法研究泛素的分布。 结果 TGF - β1,2 ,3mRNA用原位杂交方法均能检测到 ,但它们分布的位置不同。TGF - β1mRNA仅存在于人工腱周围新生的腱成纤维细胞质中。TGF - β2mRNA在新生腱组织中广泛存在 ,在半成熟的腱组织中也有一定程度的表达 ;而TGF - β3mRNA在新生腱组织和较成熟腱组织中表达量很低。在正常自体腱组织中 ,泛素呈低表达 ,且均匀分布 ;HHK -Ⅱ人工腱植入后 3周 ,人工腱之间及其周围的新生腱组织中泛素表现为强阳性 ;植入后 6周和 9周的泛素为弱阳性 ,其中 9周的泛素接近正常腱组织。即在人工腱降解期泛素表现为强阳性 ,自体腱形成期及之后的时期泛素的表达明显下调。 结论 在肌腱修复时 ,TGF - β1,2可能起主要作用 ,TGF - β3在肌腱修复过程中起协同作用。人工腱角蛋白的降解可能主要由泛素承担 ;人工腱周围新生组织中不正常蛋白的清除除有溶酶体参与外 ,泛素也?
Objective To study the expression of transforming growth factor - β1, 2, 3 (TGF - β1, 2, 3) mRNA and the expression of ubiquitin mRNA in the second generation human hairkeratin Ⅱ (HHK - Ⅱ) Distribution. Methods Four New Zealand rabbits were used to make animal models. The expressions of TGF - β1, 2 and 3 mRNA in different stages of HHK - Ⅱ artificial tendon implantation were detected by in situ hybridization (ISH). The expression of ubiquitin Distribution. Results TGF - β1, 2 and 3 mRNAs were all detected by in situ hybridization method, but their locations were different. TGF - β1 mRNA is present only in the nascent tendon fibroblast cytoplasm around the artificial tendon. TGF - β2 mRNA is widely expressed in new tendon tissues and also expressed to a certain extent in semi - mature tendon tissues. However, TGF - β3 mRNA expression is low in new tendon tissues and mature tendon tissues. In normal autologous tendon tissue, ubiquitin showed low expression and was evenly distributed; UHU tended to be strongly positive in the new tendon tissue between and around the artificial HHK-II artificial tendon 3 weeks after implantation Ubiquitin was weakly positive at weeks 6 and 9, with 9 weeks of ubiquitin near normal tendon tissue. That is to say, ubiquitin showed strong positive during the degradation of artificial tendon and the expression of ubiquitin was significantly down-regulated during and after autologous tendon formation. Conclusion TGF - β1,2 may play a major role in tendon repair. TGF - β3 may play a synergistic role in tendon repair. Degradation of artificial tendon keratin may be mainly assumed by ubiquitin; Artificial tendon neoplasm abnormal protein removal in addition to the involvement of lysosomes, ubiquitin also?