【摘要】：骨性關(guān)節(jié)炎及其晚期出現(xiàn)的骨軟骨缺損等尚無理想的治療方法,影響著人們?nèi)粘Ｉ詈凸ぷ�。骨性關(guān)節(jié)炎早期出現(xiàn)軟骨病變的臨床藥物均是對癥處理,主要原因是目前關(guān)節(jié)透明軟骨生理和病理知識缺乏。骨性關(guān)節(jié)炎晚期所致的骨軟骨缺損修復,既需要糾正其病理機制又需要提供再生的材料。臨床上用于神經(jīng)系統(tǒng)退行性疾病和腫瘤疾病治療的重要信號通路靶標,均已經(jīng)過系統(tǒng)基礎(chǔ)和臨床驗證。骨性關(guān)節(jié)炎病理機制具有與之相似的細胞內(nèi)信號通路改變,因此這些系統(tǒng)的臨床藥物是開發(fā)軟骨疾病的良好候選藥物,是控制骨性關(guān)節(jié)炎早期軟骨破壞和促進后期骨軟骨缺損再生的理想小分子。 本課題以小分子藥物吉非替尼和氯化鋰為主要研究對象,分別對關(guān)節(jié)腔內(nèi)緩釋吉非替尼預(yù)防骨性關(guān)節(jié)炎早期軟骨破壞的作用和機制以及構(gòu)建緩釋鋰離子介孔生物活性玻璃支架促進骨性關(guān)節(jié)炎晚期骨軟骨缺損再生的作用和機制進行了相關(guān)研究。 在對關(guān)節(jié)腔內(nèi)緩釋吉非替尼預(yù)防骨性關(guān)節(jié)炎早期軟骨破壞的機制研究中發(fā)現(xiàn)磷酸化EGFR在骨性關(guān)節(jié)炎的軟骨中表達升高,并伴隨軟骨自噬水平的降低。同時,在使用TGF-a構(gòu)建的軟骨細胞關(guān)節(jié)炎模型中觀察到相同的結(jié)果。TGF-a導致的關(guān)節(jié)炎效應(yīng)可以被EGFR的小干擾RNA或該受體的絡(luò)氨酸激酶抑制劑吉非替尼阻斷,繼而增加膠原基因的表達和抑制金屬降解酶MMP13的表達,且該效應(yīng)通過自噬關(guān)鍵基因ATG5發(fā)揮作用。為了進一步探討吉非替尼的體內(nèi)效應(yīng),向骨性關(guān)節(jié)炎小鼠關(guān)節(jié)腔內(nèi)注射吉非替尼殼聚糖緩釋微球。結(jié)果顯示：與對照組相比,含藥微球維持了軟骨的內(nèi)平衡并同時抑制了EGFR受體的表達,吉非替尼殼聚糖微球增加了膠原的表達和抑制了降解酶MMP13的表達,并伴隨自噬關(guān)鍵蛋白Beclin1和LC3的表達。 緩釋鋰離子介孔生物活性玻璃支架促進骨軟骨缺損修復的研究中發(fā)現(xiàn)：在兔子骨軟骨缺損模型中,與對照組相比,含鋰介孔介孔生物活性玻璃支架促進了軟骨下骨和透明軟骨的再生。進一步研究表明支架中釋放的鋰離子在支架促進缺損再生中起關(guān)鍵作用,鋰離子通過激活軟骨下骨中骨髓間充質(zhì)干細胞的Wnt信號通路促進其增殖和向骨系分化,同時鋰離子通過提高軟骨的自噬水平抑制骨性關(guān)節(jié)炎的炎癥環(huán)境而發(fā)揮對軟骨的保護作用。 以上結(jié)果表明關(guān)節(jié)腔緩釋吉非替尼通過EGFR-自噬軸延緩小鼠骨性關(guān)節(jié)炎的進展,因此,提示吉非替尼可以作為骨性關(guān)節(jié)炎患者的臨床候選藥物；含鋰介孔生物活性玻璃支架通過同時模擬軟骨下骨和軟骨的生物學特征促進骨軟骨缺損的再生。
[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.
【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：R684.3

【參考文獻】

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1 花芳;余嬌嬌;李珂;胡卓偉;;自噬影響衰老及老年病的研究進展[J];藥學學報;2014年06期

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本文編號：2415260