吉非替尼和锂盐治疗骨性关节炎的效应和机制研究
发布时间：2019-01-26 07:27
【摘要】：骨性关节炎及其晚期出现的骨软骨缺损等尚无理想的治疗方法,影响着人们日常生活和工作。骨性关节炎早期出现软骨病变的临床药物均是对症处理,主要原因是目前关节透明软骨生理和病理知识缺乏。骨性关节炎晚期所致的骨软骨缺损修复,既需要纠正其病理机制又需要提供再生的材料。临床上用于神经系统退行性疾病和肿瘤疾病治疗的重要信号通路靶标,均已经过系统基础和临床验证。骨性关节炎病理机制具有与之相似的细胞内信号通路改变,因此这些系统的临床药物是开发软骨疾病的良好候选药物,是控制骨性关节炎早期软骨破坏和促进后期骨软骨缺损再生的理想小分子。 本课题以小分子药物吉非替尼和氯化锂为主要研究对象,分别对关节腔内缓释吉非替尼预防骨性关节炎早期软骨破坏的作用和机制以及构建缓释锂离子介孔生物活性玻璃支架促进骨性关节炎晚期骨软骨缺损再生的作用和机制进行了相关研究。 在对关节腔内缓释吉非替尼预防骨性关节炎早期软骨破坏的机制研究中发现磷酸化EGFR在骨性关节炎的软骨中表达升高,并伴随软骨自噬水平的降低。同时,在使用TGF-a构建的软骨细胞关节炎模型中观察到相同的结果。TGF-a导致的关节炎效应可以被EGFR的小干扰RNA或该受体的络氨酸激酶抑制剂吉非替尼阻断,继而增加胶原基因的表达和抑制金属降解酶MMP13的表达,且该效应通过自噬关键基因ATG5发挥作用。为了进一步探讨吉非替尼的体内效应,向骨性关节炎小鼠关节腔内注射吉非替尼壳聚糖缓释微球。结果显示：与对照组相比,含药微球维持了软骨的内平衡并同时抑制了EGFR受体的表达,吉非替尼壳聚糖微球增加了胶原的表达和抑制了降解酶MMP13的表达,并伴随自噬关键蛋白Beclin1和LC3的表达。 缓释锂离子介孔生物活性玻璃支架促进骨软骨缺损修复的研究中发现：在兔子骨软骨缺损模型中,与对照组相比,含锂介孔介孔生物活性玻璃支架促进了软骨下骨和透明软骨的再生。进一步研究表明支架中释放的锂离子在支架促进缺损再生中起关键作用,锂离子通过激活软骨下骨中骨髓间充质干细胞的Wnt信号通路促进其增殖和向骨系分化,同时锂离子通过提高软骨的自噬水平抑制骨性关节炎的炎症环境而发挥对软骨的保护作用。 以上结果表明关节腔缓释吉非替尼通过EGFR-自噬轴延缓小鼠骨性关节炎的进展,因此,提示吉非替尼可以作为骨性关节炎患者的临床候选药物；含锂介孔生物活性玻璃支架通过同时模拟软骨下骨和软骨的生物学特征促进骨软骨缺损的再生。
[Abstract]:There is no ideal treatment for osteoarthritis and its late stage osteochondral defect, which affects people's daily life and work. The main reason for the early occurrence of osteoarthritis is the lack of physiological and pathological knowledge of articular hyaline cartilage. The repair of osteochondral defects caused by late osteoarthritis requires both correction of its pathological mechanism and provision of regenerative materials. The important signal pathway targets for the treatment of neurodegenerative diseases and tumor diseases have been systemically and clinically verified. The pathological mechanism of osteoarthritis has similar changes in intracellular signaling pathways, so the clinical drugs of these systems are good candidates for developing cartilage diseases. It is an ideal small molecule for controlling early cartilage destruction and promoting regeneration of osteochondral defect in the later stage of osteoarthritis. In this paper, the small molecular drugs gefitinib and lithium chloride as the main research object, Effects and mechanisms of intraarticular sustained-release gefitinib on the prevention of early cartilage destruction in osteoarthritis and the effects of constructing lithium-ion bioactive glass scaffolds on regeneration of osteochondral defects in the late stage of osteoarthritis And the mechanism has been studied. It was found that the expression of phosphorylated EGFR in the cartilage of osteoarthritis was increased and the level of cartilage autophagy was decreased. At the same time, the same results were observed in the chondrocyte arthritis model constructed with TGF-a. The arthritis effect induced by TGF-a could be blocked by the small interfering RNA of EGFR or the receptor's complex kinase inhibitor gefitinib. The expression of collagen gene and the expression of metal-degrading enzyme MMP13 were inhibited, and the effect was played by autophagy key gene ATG5. In order to further study the in vivo effect of gefitinib, gifetini chitosan sustained release microspheres were injected into the articular cavity of osteoarthritis mice. The results showed that: compared with the control group, the drug containing microspheres maintained the internal balance of cartilage and inhibited the expression of EGFR receptor, and gefitinib chitosan microspheres increased the expression of collagen and inhibited the expression of degrading enzyme MMP13. The expression of Beclin1 and LC3 was associated with autophagy. In the study of slow-release lithium ion mesoporous bioactive glass scaffolds promoting the repair of bone cartilage defect, it was found that in rabbit bone cartilage defect model, compared with the control group, Lithium-containing mesoporous bioactive glass scaffolds promote the regeneration of subchondral bone and hyaline cartilage. Further studies have shown that lithium ions released in the scaffold play a key role in promoting the regeneration of the scaffold. Lithium ions promote the proliferation and differentiation of bone marrow mesenchymal stem cells by activating the Wnt signaling pathway of bone marrow mesenchymal stem cells in the subchondral bone. At the same time, lithium ion plays a protective role on cartilage by increasing the autophagy level of cartilage and inhibiting the inflammatory environment of osteoarthritis. These results suggest that the joint cavity sustained-release gefitinib delays the progression of osteoarthritis through the EGFR- autophagy axis. Therefore, it is suggested that gefitinib can be used as a clinical candidate for patients with osteoarthritis. Lithium-containing mesoporous bioactive glass scaffolds promote the regeneration of bone defects by simultaneously simulating the biological characteristics of subchondral bone and cartilage.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R684.3
本文编号：2415260
[Abstract]:There is no ideal treatment for osteoarthritis and its late stage osteochondral defect, which affects people's daily life and work. The main reason for the early occurrence of osteoarthritis is the lack of physiological and pathological knowledge of articular hyaline cartilage. The repair of osteochondral defects caused by late osteoarthritis requires both correction of its pathological mechanism and provision of regenerative materials. The important signal pathway targets for the treatment of neurodegenerative diseases and tumor diseases have been systemically and clinically verified. The pathological mechanism of osteoarthritis has similar changes in intracellular signaling pathways, so the clinical drugs of these systems are good candidates for developing cartilage diseases. It is an ideal small molecule for controlling early cartilage destruction and promoting regeneration of osteochondral defect in the later stage of osteoarthritis. In this paper, the small molecular drugs gefitinib and lithium chloride as the main research object, Effects and mechanisms of intraarticular sustained-release gefitinib on the prevention of early cartilage destruction in osteoarthritis and the effects of constructing lithium-ion bioactive glass scaffolds on regeneration of osteochondral defects in the late stage of osteoarthritis And the mechanism has been studied. It was found that the expression of phosphorylated EGFR in the cartilage of osteoarthritis was increased and the level of cartilage autophagy was decreased. At the same time, the same results were observed in the chondrocyte arthritis model constructed with TGF-a. The arthritis effect induced by TGF-a could be blocked by the small interfering RNA of EGFR or the receptor's complex kinase inhibitor gefitinib. The expression of collagen gene and the expression of metal-degrading enzyme MMP13 were inhibited, and the effect was played by autophagy key gene ATG5. In order to further study the in vivo effect of gefitinib, gifetini chitosan sustained release microspheres were injected into the articular cavity of osteoarthritis mice. The results showed that: compared with the control group, the drug containing microspheres maintained the internal balance of cartilage and inhibited the expression of EGFR receptor, and gefitinib chitosan microspheres increased the expression of collagen and inhibited the expression of degrading enzyme MMP13. The expression of Beclin1 and LC3 was associated with autophagy. In the study of slow-release lithium ion mesoporous bioactive glass scaffolds promoting the repair of bone cartilage defect, it was found that in rabbit bone cartilage defect model, compared with the control group, Lithium-containing mesoporous bioactive glass scaffolds promote the regeneration of subchondral bone and hyaline cartilage. Further studies have shown that lithium ions released in the scaffold play a key role in promoting the regeneration of the scaffold. Lithium ions promote the proliferation and differentiation of bone marrow mesenchymal stem cells by activating the Wnt signaling pathway of bone marrow mesenchymal stem cells in the subchondral bone. At the same time, lithium ion plays a protective role on cartilage by increasing the autophagy level of cartilage and inhibiting the inflammatory environment of osteoarthritis. These results suggest that the joint cavity sustained-release gefitinib delays the progression of osteoarthritis through the EGFR- autophagy axis. Therefore, it is suggested that gefitinib can be used as a clinical candidate for patients with osteoarthritis. Lithium-containing mesoporous bioactive glass scaffolds promote the regeneration of bone defects by simultaneously simulating the biological characteristics of subchondral bone and cartilage.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R684.3
【参考文献】
相关期刊论文 前1条
1 花芳;余娇娇;李珂;胡卓伟;;自噬影响衰老及老年病的研究进展[J];药学学报;2014年06期
，本文编号：2415260
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