核心蛋白聚糖对声带瘢痕治疗的初步研究

发布时间：2018-04-24 09:28

本文选题：核心蛋白聚糖 + 声带瘢痕　；参考：《山东中医药大学》2017年硕士论文

【摘要】：目的:构建新西兰兔声带损伤模型,在声带瘢痕形成的不同时期,于损伤部位周围多方向外源性注入核心蛋白聚糖(Decorin,DCN),分析声带组织病理学改变和转化生长因子-β1(Transforming growth factor-β1,TGF-β1)表达变化,观察DCN能否抑制声带瘢痕增生,探讨DCN抑制瘢痕增生的作用机制。方法:选取新西兰兔25只,随机选取20只做双侧声带急性损伤模型,用喉钳夹取双侧声带前中部组织。选取10只新西兰兔立即进行右侧声带损伤部位周围多方向注入100μg/L的核心蛋白聚糖,左侧声带损伤部位作为对照声带(A组),其它10只于声带损伤1月后进行右侧声带损伤部位周围多方向注入100μg/L的核心蛋白聚糖,左侧声带损伤部位作为对照声带(B组),剩余5只新西兰兔作为无干预对照组(C组)。A组、B组新西兰兔于注射核心蛋白聚糖后4周、12周分别随机挑选5只收获声带组织。将3组新西兰兔收获的声带组织进行HE染色和免疫组织化学染色,分析声带组织的病理学改变和TGF-β1表达变化,将各组所得值相对比并进行统计学分析。结果:1、通过环甲膜切开的方式损伤新西兰兔的声带组织,成功完成了声带损伤动物模型的构建,为接下来研究外源性注入DCN能否抑制声带瘢痕增生奠定了基础。2、HE染色结果新西兰兔的无损伤声带组织可见明显的分层构造,依次为上皮层、固有层和肌层,固有层主要是松散的结缔组织,胶原纤维的排列比较规则。声带损伤组可见声带固有层的纤维组织成分明显增多,胶原纤维比较密集,排列也非常杂乱,染色比无损伤声带明显加深。DCN注射组可见固有层纤维组织比无损伤声带有所增多,但胶原纤维的排列比较规则,染色比声带损伤组明显变浅。3、免疫组织化学染色结果(1)A、B两组比C组声带TGF-β1的阳性表达高,有显著的统计学意义。(P0.01)(2)A、B两组右侧声带比左侧声带TGF-β1的阳性表达明显减少,差异有显著统计学意义。(P0.01)(3)A组比B组TGF-β1的阳性表达明显减少,差异有统计学意义。(P0.01)(4)A、B两组,12周后观察组比4周后观察组TGF-β1的阳性表达无明显变化,差异没有统计学意义。(P=0.17、P=0.14,P0.05)结论:1、DCN能够与胶原纤维高度亲和并以此来调节其形成,使其在组织中的表达受到干扰,稳定性变差,进而改变其装配和结构,从而抑制瘢痕增生。2、TGF-β1是介导纤维增生性疾病的关键因子,DCN能够通过结合TGF-β1,并且灭活TGF-β1的生物活性,来抑制瘢痕形成。瘢痕形成的早期纤维化程度比较轻,早期注射效果比较好,而且DCN降解速度适当,是良好的防治瘢痕的材料。
[Abstract]:Objective: to construct the vocal cord injury model of New Zealand rabbits, and to analyze the pathological changes of vocal cord and the expression of transforming growth factor-尾 1(Transforming factor- 尾 1 (TGF- 尾 1). To observe whether DCN can inhibit vocal cord scar proliferation and to explore the mechanism of DCN inhibiting scar hyperplasia. Methods: 25 New Zealand rabbits were selected and 20 rabbits were randomly selected to make the model of bilateral vocal cord acute injury. The anterior and middle tissues of bilateral vocal cord were clamped by larynx clamp. Ten New Zealand rabbits were immediately injected with 100 渭 g / L core proteoglycan around the injured site of the right vocal cord. The left vocal cord injury site was used as the control vocal cord group A, the other 10 rats were injected with 100 渭 g / L core proteoglycan around the right vocal cord injury site one month after the vocal cord injury, and the other 10 rats were injected with 100 渭 g / L core proteoglycan around the right vocal cord injury site. The injured sites of left vocal cord were used as control vocal cord group B, and the remaining 5 New Zealand rabbits as control group C, group A and group B were randomly selected to harvest vocal cord tissue 4 weeks and 12 weeks after injection of core proteoglycan. The vocal cord tissues harvested from three groups of New Zealand rabbits were stained with HE and immunohistochemistry. The pathological changes of vocal cords and the expression of TGF- 尾 1 were analyzed, and the values of each group were compared and statistically analyzed. Results the vocal cord tissue of New Zealand rabbits was damaged by cyclidine incision, and the animal model of vocal cord injury was successfully constructed. In order to study whether exogenous injection of DCN can inhibit vocal cord scar proliferation, the results of HE staining showed that the undamaged vocal cord tissue in New Zealand rabbits was obviously stratified in order of epithelium, lamina propria and muscularis. The lamina propria is mainly loose connective tissue, and the arrangement of collagen fibers is regular. In the vocal cord injury group, the fibrous tissue of the lamina propria of vocal cord was obviously increased, the collagen fiber was dense, and the arrangement was very messy. The staining of the lamina propria was significantly deeper than that of the non-injuried vocal cord. The fibrous tissue of the lamina propria was increased in the group of DCN injection than that in the non-injured vocal cord. But the arrangement of collagen fibers was regular, the staining was lighter than that of vocal cord injury group, and the positive expression of TGF- 尾 1 was higher in two groups than that in C group, and the expression of TGF- 尾 1 in two groups was higher than that in C group, and the expression of TGF- 尾 1 in group B was higher than that in group C. The positive expression of TGF- 尾 1 in the right vocal cord was significantly lower than that in the left vocal cord, and the difference was statistically significant. The positive expression of TGF- 尾 1 in the right vocal cord group was significantly lower than that in the B group, and the positive expression of TGF- 尾 1 in the right vocal cord group was significantly lower than that in the left vocal cord group. The difference was statistically significant. The positive expression of TGF- 尾 1 in the observation group after 12 weeks was not significantly different from that in the observation group after 4 weeks. The difference was not statistically significant (P < 0. 17, P < 0. 14, P 0. 05). Conclusion: 1. The expression of TGF- 尾 _ 1 in tissues is disturbed, its stability is deteriorated, and its assembly and structure are changed, thus inhibiting scar proliferation. TGF- 尾 _ 1 is a key factor in mediating fibrous hyperplastic disease. DCN can inhibit the biological activity of TGF- 尾 _ 1 by binding to TGF- 尾 _ 1, and inactivating the bioactivity of TGF- 尾 _ 1. To inhibit scar formation. The degree of early fibrosis of scar formation is relatively light, the effect of early injection is better, and the DCN degradation rate is appropriate, so it is a good material for the prevention and treatment of scar.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R767.4

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