严斌等

[摘要] 目的 探讨茶多酚对大鼠骨关节软骨氧化损伤的保护作用，以进一步了解茶多酚摄入机体对骨折早期愈合的影响机制。 方法 选取3～4月龄健康雄性Sprague-Dawley大鼠45只作为观察对象，随机将其分为A、B、C三组，每组15只。各组均制造骨折前茶多酚生物效应，对三组大鼠进行皮下注射，A组剂量为5 mg/（kg·d）茶多酚，B组剂量为10 mg/（kg·d）茶多酚，而C组剂量为0.5 mL/（kg·d）生理盐水；注射3周后，将每只大鼠桡骨中下1/3交界处造成左侧桡骨3 mm完全骨质缺损的骨折模型，不固定；骨折后继续注射茶多酚，制造骨折后茶多酚生物效应；采用HE染色分别观察大鼠在骨折愈合不同时期（造模后3、7、14、21 d）的骨痂厚度及骨痂成熟度。 结果 ①骨痂厚度：经HE染色观察，造模后3 d，三组间比较差异无统计学意义（P > 0.05）；造模后7 d，B组明显高于其他两组，差异有统计学意义（P < 0.05）；造模后14 d， A组与B组明显高于C组，差异有统计学意义（P < 0.05）；造模后21 d，B组明显高于A组与C组，且A组明显高于C组，各组间差异均有统计学意义（P < 0.05）。②A组与B组大鼠术后3 d即可见到骨内外膜开始增厚，术后7 d骨折端肉芽组织形成，术后14 d骨折端肉芽组织开始重新生长，术后21 d骨折端纤维性骨痂与纤维母细胞有明显减少；C组术后3、7 d骨内外膜也有所增厚，但是与术前相比差异不明显，术后14、21 d骨痂中纤维细胞、毛细血管密度及纤维性结缔组织有明显增加，出现新生血管，但是成骨细胞的增殖数量与A、B组相比明显偏低，而且骨痂的成熟度也相对较低，内外骨痂改造塑形期出现较晚。 结论 茶多酚对骨关节软骨氧化损伤具有明显的保护作用，其可以通过促进骨痂增殖分化，增加骨痂含量，加快骨痂的成熟，从而提高骨折的愈合速度。

[关键词] 茶多酚；骨关节软骨；氧化损伤；保护；肿瘤坏死因子

[中图分类号] R285 [文献标识码] A [文章编号] 1673-7210（2015）01（b）-0016-04

[Abstract] Objective To explore the protection of tea polyphenols on oxidative injury of the rat articular cartilages to further understand the effect of intake of tea polyphenols on the early healing of the fracture. Methods 45 healthy male Sprague-Dawley rats which were 3-4 month old were selected as the research objects and randomly divided into A， B， C three groups， 15 rats in each group. The rats in each group manufactured biological effects of tea polyphenols before fracture and the rats in the three groups received subcutaneous injection. The rats in group A were injected 5 mg/（kg·d） of tea polyphenols， rats in group B were injected 10 mg/（kg·d） of tea polyphenols， while rats in group C were injected 0.5 mL/（kg·d） physiological brine. Three weeks after the injection， the fracture model of each rat middle radius 1/3 junction caused by left radius 3 mm complete bone defectis were not fixed； after fracture continue to inject tea polyphenols， tea polyphenols manufacturing fracture afterbiological effects were observed by HE staining； rats in different period of fracture healing （model after 3， 7， 14， 21 d） of bone thickness and bone callus maturity. Results ①Callus thickness： observed by HE staining， 3 d after modeling， significant comparison between three groups showed that， differences were not statistically significant （P > 0.05）； 7 d after modeling， group B was significantly higher than the other two groups， differences had statistical significance （P < 0.05）； 14 d after modeling， group A and group B was significantly higher than that in group C， differences were statistical significance （P < 0.05）； 21 d after modeling， group B was significantly higher than that in group A and group C， and group A was significantly higher than that in group C， differences were statistical significance （P < 0.05）. ②3 d after the operation in group A and group B rats can be seen in the outer membrane of bonethickens， 7 d after operation fracture end of granulation tissue formation， 14 d after operation fracture end of granulation tissue growing back， after 21 d fracture of fibrous callus and fibroblast were significantly reduced in group C； 3， 7 d after operation， bone in outer membrane had thickened， but the difference was not significant compared with the preoperative， postoperativefiber 14， 21 d in callus cells， capillary density and fibrous connective tissue increased significantly， neovascularization， but the osteoblast proliferationcompared with group A， group B was significantly lower， and the callus maturity is relatively low， internal， external callus transformation and shaping period appeared later. Conclusion Tea polyphenols has a significant protective effect on articular cartilage of oxidative damage， which canpromote the proliferation and differentiation of the callus， callus content increase， accelerate callus of mature， so improve the fracture healing rate.

[Key words] Tea polyphenols； Articular cartilage； Oxidative damage； Protection； Tumor necrosis factor

'骨折后形成的骨痴组织对骨的修复、愈合过程至关重要，也是骨折愈合程度的评定标准[1-2]。茶多酚对机体细胞和组织具有复杂的生物效应，本课题拟采用动物实验方法研究茶多酚对大鼠骨折愈合的影响，通过建立茶多酚生物效应下的骨折愈合模型，使用HE染色法检测骨痂厚度及成熟度，使用免疫组化染色法检测骨痂中肿瘤坏死因子（TNF-α）表达水平来探讨茶多酚对骨折愈合的早期影响。

1 对象与方法

1.1 研究对象

选取3～4月龄健康雄性Sprague-Dawley（SD）大鼠45只，作为本组研究的观察对象，随机将其分为A、B、C三组，每组15只。入……