本文選題：整合素 + 骨關(guān)節(jié)炎　； 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：骨關(guān)節(jié)炎（Osteoarthritis，OA）是一種退行性關(guān)節(jié)疾病，主要病理表現(xiàn)為關(guān)節(jié)軟骨的退變，好發(fā)于中老年，膝關(guān)節(jié)常受累。隨著世界人口老齡化進(jìn)程的加速，OA的發(fā)病率逐年上升，已成為中老年人致殘的重要原因之一，給患者帶來(lái)極大痛苦與經(jīng)濟(jì)負(fù)擔(dān)，嚴(yán)重影響患者的生活質(zhì)量。由于OA的病因與發(fā)病機(jī)制尚不清楚，目前仍是骨科領(lǐng)域研究的熱點(diǎn)與難點(diǎn)。研究發(fā)現(xiàn)，整合素（Integrin）是細(xì)胞粘附分子家族的一員，存在于細(xì)胞表面，是一種跨膜蛋白，在細(xì)胞粘附與信號(hào)傳導(dǎo)中起著重要作用，參與細(xì)胞形態(tài)、極性、粘附、運(yùn)動(dòng)、生長(zhǎng)、增殖、分化與凋亡等多種生理功能的調(diào)節(jié)。軟骨細(xì)胞可表達(dá)多種Integrins，成人軟骨細(xì)胞中主要表達(dá)Integrinα5β1。Integrinα5β1在軟骨細(xì)胞力學(xué)信號(hào)轉(zhuǎn)導(dǎo)、細(xì)胞增殖與分化等方面具有重要作用。研究發(fā)現(xiàn)，Integrinα5β1與軟骨細(xì)胞的損傷、修復(fù)、細(xì)胞表型改變等多種軟骨病變密切相關(guān)。我們推測(cè)，Integrinα5β1在不同退變程度軟骨中可能存在差異性表達(dá)，且與軟骨的退變及OA的發(fā)展相關(guān)。目前，關(guān)于Integrinα5β1在不同退變程度軟骨中的表達(dá)情況及與軟骨退變的關(guān)系缺乏相關(guān)研究。本實(shí)驗(yàn)選取膝骨關(guān)節(jié)炎（Knee Osteoarthritis，KOA）患者股骨遠(yuǎn)端負(fù)重面不同退變程度關(guān)節(jié)軟骨，檢測(cè)其中Integrinα5β1的表達(dá)，探討Integrinα5β1在KOA不同退變程度軟骨中的表達(dá)情況及與軟骨退變的關(guān)系。方法：收集KOA患者全膝關(guān)節(jié)置換術(shù)股骨遠(yuǎn)端截骨組織20例，經(jīng)大體觀察，依據(jù)Outerbridge分級(jí)標(biāo)準(zhǔn)選取不同退變程度軟骨標(biāo)本共40例（每例標(biāo)本沿矢狀面等分成三塊，一塊用10%中性甲醛固定，另兩塊－80℃凍藏保存），將10%中性甲醛固定的標(biāo)本脫鈣后制成石蠟組織塊，3μm切片后，常規(guī)HE染色及鏡下觀察，參照Bobinac分期，將標(biāo)本分為三組：輕度退變組（0～6分）、中度退變組（7～9分）、重度退變組（10～14分）。采用免疫組化、Real-time PCR及Western blot檢測(cè)Integrinα5β1在三組不同退變程度軟骨中mRNA與蛋白水平的表達(dá)。實(shí)驗(yàn)數(shù)據(jù)采用均數(shù)±標(biāo)準(zhǔn)差表示，統(tǒng)計(jì)分析采用SPSS17.0軟件，三組間比較采用單因素方差分析，P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果：本實(shí)驗(yàn)20例KOA患者股骨遠(yuǎn)端截骨順利，內(nèi)外髁負(fù)重面軟骨無(wú)人為損傷及污染，共選取軟骨標(biāo)本40例，HE染色均勻、紅藍(lán)分明、組織結(jié)構(gòu)清晰、胞漿胞核清晰可辨、鏡下觀察與判別良好，參照Bobinac分期分成三組，輕度退變組13例、中度退變組15、重度退變組12例。免疫組化染色顯示：Integrinα5β1在不同退變程度軟骨中均有表達(dá)；輕度退變組主要表達(dá)在軟骨淺表層與移形層；中度退變組除鈣化層周圍軟骨細(xì)胞未見明顯表達(dá)外，其余軟骨細(xì)胞均有表達(dá)，侵及軟骨潮線的血管內(nèi)皮細(xì)胞可見表達(dá)；重度退變組所有軟骨細(xì)胞與侵入軟骨的血管內(nèi)皮細(xì)胞均有表達(dá)。輕度退變組呈低表達(dá)，中度退變組表達(dá)增強(qiáng)，重度退變組表達(dá)明顯增強(qiáng)；免疫組化檢測(cè)結(jié)果經(jīng)Image-Pro Plus6.0軟件半定量分析，三組間比較，差異性顯著（P＜0.05）。Real-time PCR與Western blot檢測(cè)顯示：Integrinα5β1的mRNA與蛋白在三組不同退變程度軟骨中均有表達(dá)，兩種方法檢測(cè)結(jié)果相同，輕度退變組呈低表達(dá)，中度退變組表達(dá)增強(qiáng)，，重度退變組表達(dá)明顯增強(qiáng)；Real-time PCR與Western blot檢測(cè)結(jié)果對(duì)應(yīng)經(jīng)ABI StepOne PlusReal-time PCR System與Quantity One軟件定量分析，同種方法的三組間比較，差異性顯著（P＜0.05）。結(jié)論：Integrinα5β1在KOA不同退變程度軟骨中均有表達(dá)，各組間的表達(dá)存在差異性；隨著軟骨退變程度的加重，Integrinα5β1在軟骨中的表達(dá)由淺表層細(xì)胞逐漸擴(kuò)展到全層軟骨細(xì)胞，表達(dá)量逐漸增強(qiáng)。Integrinα5β1在不同退變程度軟骨中的差異性表達(dá)與軟骨退變及OA的發(fā)展具有密切聯(lián)系。
[Abstract]:Objective: Osteoarthritis (OA) is a degenerative joint disease. The main pathological manifestation is the degeneration of articular cartilage. The knee joint is often affected in the middle and old age. With the acceleration of the aging process of the world population, the incidence of OA has risen year by year. It has become one of the important reasons for the disabled people in the middle and old years, which brings great pain to the patients. And the economic burden seriously affects the quality of life of the patients. Because the etiology and pathogenesis of OA is still unclear, it is still a hot and difficult point in the field of Department of orthopedics. It is found that integrin (Integrin) is a member of the cell adhesion molecule family, which exists on the cell surface, is a kind of transmembrane protein, and plays an important role in cell adhesion and signal transduction. To participate in the regulation of many physiological functions, such as cell morphology, polarity, adhesion, movement, growth, proliferation, differentiation and apoptosis. Chondrocytes can express a variety of Integrins. The main expression of Integrin alpha 5 beta 1.Integrin alpha 5 beta 1 in adult chondrocytes is important in the study of chondrocyte mechanical signal transduction, cell proliferation and differentiation. It is found that Integrin alpha 5 beta 1 is closely related to cartilage damage, repair, cell phenotype change and so on. We speculate that Integrin alpha 5 beta 1 may have differential expression in different degeneration cartilage, and is related to cartilage degeneration and the development of OA. At present, the expression of Integrin alpha 5 beta 1 in cartilage of different degeneration degree The relationship between the condition and the cartilage degeneration is not related. This experiment selected the articular cartilage of the distal femur with different degrees of degeneration in Knee Osteoarthritis (KOA). The expression of Integrin alpha 5 beta 1 was detected, and the expression of Integrin alpha 5 beta 1 in the cartilage of different degree of degeneration of KOA and the relationship with cartilage degeneration were investigated. Methods: 20 cases of distal femur osteotomy in total knee arthroplasty of KOA patients were collected, and 40 cases with different degree of degeneration were selected according to the standard of Outerbridge classification. Three pieces were divided into three blocks along the sagittal plane, one was fixed with 10% neutral formaldehyde, and the other two - 80 centigrade frozen storage was preserved. The target of 10% neutral formaldehyde was fixed. After decalcification, the paraffin tissue was made. After 3 m slices, conventional HE staining and microscopic observation were used. The specimens were divided into three groups: mild degeneration group (0~6), moderate degeneration group (7~9) and severe degeneration group (10~14 points). Immunohistochemistry, Real-time PCR and Western blot were used to detect Integrin alpha 5 beta 1 in three groups of different degrees of degeneration soft The expression of mRNA and protein in bone was expressed. The experimental data were expressed with mean standard deviation, and SPSS17.0 software was used for statistical analysis. The three groups were compared with single factor analysis of variance. The difference was statistically significant in P < 0.05. Results: the distal femur osteotomy was smooth in 20 cases of KOA patients and no one was damaged and polluted by the internal and external condyle bearing face cartilage. 40 cases of cartilage specimens were selected, HE staining was uniform, red and blue, clear tissue structure, clear differentiation of cytoplasm nucleus, good observation and discrimination under the microscope, Bobinac staging was divided into three groups, 13 cases in mild degeneration group, 15 in moderate degeneration group and 12 in severe degeneration group. Immunohistochemical staining showed that Integrin alpha 5 beta 1 was expressed in different degeneration degree cartilage The mild degeneration group was mainly expressed in the superficial cartilaginous surface and the transitional layer, and the other chondrocytes were expressed in the moderate degeneration group except the cartilage cells in the calcification layer, and the vascular endothelial cells that invaded the cartilage tidal line were expressed, and all the cartilage cells in the severe degeneration group were expressed in the vascular endothelial cells that invaded the cartilage. The expression of mild degeneration group was low, the expression of moderate degenerative group was enhanced and the expression of severe degeneration group was obviously enhanced. The results of immunohistochemical detection were semi quantitative analysis by Image-Pro Plus6.0 software. The difference was significant between the three groups (P < 0.05).Real-time PCR and Western blot detection: the mRNA and protein of Integrin alpha 5 beta 1 were in three groups of different degenerative processes. The results of the two methods were the same, the results of the two methods were the same, the mild degeneration group showed low expression, the moderate degeneration group was enhanced, the expression of the severe degeneration group was obviously enhanced; the results of Real-time PCR and Western blot were quantified by ABI StepOne PlusReal-time PCR System and Quantity One software, and compared between the same methods. The difference was significant (P < 0.05). Conclusion: the expression of Integrin alpha 5 beta 1 was expressed in different degree of degeneration of KOA cartilage, and the expression in each group was different. With the aggravation of cartilage degeneration, the expression of Integrin alpha 5 beta 1 in cartilage gradually expanded from superficial layer to whole layer cartilage cell, and the expression gradually increased.Integrin alpha 5 beta 1 in different The differential expression of degeneration in cartilage is closely related to the degeneration of cartilage and the development of OA.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R684.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 林燕萍;何嘉承;佘家Y

本文編號(hào)：2032269