本文選題：膝前交叉韌帶　切入點(diǎn)：基質(zhì)金屬蛋白酶　出處：《南華大學(xué)》2013年碩士論文

【摘要】：目的：本文研究前交叉韌帶不同分束在前交叉韌帶損傷修復(fù)中的作用，探討是否前交叉韌帶的前內(nèi)側(cè)在前交叉韌帶中較后外側(cè)束作用更大。 方法：隨機(jī)抽取本院2010年6月至2012年10月具有術(shù)前MRI資料并進(jìn)行膝關(guān)節(jié)鏡探查的病例進(jìn)行回顧性分析，通過(guò)膝關(guān)節(jié)MRI矢狀位及斜矢狀位雙維掃描，檢測(cè)前交叉韌帶前內(nèi)側(cè)束、后外側(cè)束的長(zhǎng)度、厚度、寬度及脛骨止點(diǎn)之前和脛骨止點(diǎn)之后的長(zhǎng)度。在無(wú)菌環(huán)境下，從10具自愿捐贈(zèng)的新鮮尸體上取膝關(guān)節(jié)，并取出膝關(guān)節(jié)交叉前韌帶，根據(jù)ACL的解剖形態(tài)學(xué)特點(diǎn)和膝關(guān)節(jié)不同伸屈角度下的緊張狀態(tài)，將ACL分成前內(nèi)側(cè)束和后外側(cè)束兩個(gè)功能纖維束，，束間被疏松結(jié)締組織和血管分離。通過(guò)生物動(dòng)力學(xué)測(cè)試了前交叉韌帶不同部位的結(jié)構(gòu)和力學(xué)特點(diǎn)，在肌腱恢復(fù)后行拉斷試驗(yàn)，記錄負(fù)荷-延長(zhǎng)曲線，并根據(jù)此前測(cè)得的初長(zhǎng)度及橫截面積按以下公式計(jì)算最大應(yīng)變和最大應(yīng)力；提取總RNA，并反轉(zhuǎn)錄成cDNA后，應(yīng)用實(shí)時(shí)定量PCR檢測(cè)前內(nèi)側(cè)束和后外側(cè)束中與肌腱修復(fù)相關(guān)的常見MMPs基因；再通過(guò)將ACL的前內(nèi)側(cè)束和后外側(cè)束兩個(gè)功能纖維束，經(jīng)固定、石蠟包埋，利用MMP-2和MMP-3的多克隆抗體為一抗，進(jìn)行免疫組化實(shí)驗(yàn)；將前內(nèi)側(cè)束和后外側(cè)束兩個(gè)功能纖維束中的組織分別收集，勻漿后提取總蛋白，用Western blotting分析不同分束中MMP-2、MMP-3的表達(dá)量，將WB條帶進(jìn)行灰度掃描，灰度和條帶面積的乘積即為各蛋白相對(duì)表達(dá)水平。檢測(cè)了MMPs在前交叉韌帶不同部位中的表達(dá)水平。 結(jié)果： 1.通過(guò)影像學(xué)和生物動(dòng)力學(xué)測(cè)試了前交叉韌帶不同部位的結(jié)構(gòu)和力學(xué)特點(diǎn)；前內(nèi)束的最大應(yīng)力和最大應(yīng)變比后外束的大，進(jìn)一步說(shuō)明前交叉韌帶的脛骨止點(diǎn)的前部具有成為臨床肌腱重建最佳部位的潛力，為今后的臨床應(yīng)用提供理論基礎(chǔ)。 2.用RT-PCR、免疫組化、western-blot檢測(cè)了MMPs在前交叉韌帶不同部位中的表達(dá)水平，結(jié)果表明MMP-2在前內(nèi)束表達(dá)較高，在后外側(cè)束的表達(dá)較低；MMP-3在前內(nèi)側(cè)束表達(dá)較低，在后外側(cè)束的表達(dá)較高，進(jìn)一步說(shuō)明包含脛骨前端的前內(nèi)側(cè)束比后外側(cè)束更具有重塑潛力。 結(jié)論： 包含脛骨前端的前內(nèi)側(cè)束比后外側(cè)束更為重要，ACL脛骨止點(diǎn)前方更具有成為臨床肌腱修復(fù)和重塑的潛在位點(diǎn)。
[Abstract]:Aim: to study the role of different anterior cruciate ligament bundles in the repair of anterior cruciate ligament injury and to explore whether the anterior medial anterior cruciate ligament is more effective than the posterolateral bundle in the anterior cruciate ligament. Methods: from June 2010 to October 2012, patients with preoperative MRI data and knee arthroscopy were randomly selected for retrospective analysis. The anterior medial anterior cruciate ligament (ACL) anterior medial bundle was examined by MRI sagittal and oblique sagittal scan. The length, thickness, width and length of the posterolateral bundle before and after the tibial stop. Under sterile conditions, the knee joint was removed from 10 fresh cadavers donated voluntarily and the cruciate anterior ligament of the knee joint was removed. According to the anatomical morphological characteristics of ACL and the tension of knee joint at different extension and flexion angles, ACL was divided into two functional fiber bundles, the anteromedial bundle and the posterolateral bundle. The structures and mechanical characteristics of different parts of the anterior cruciate ligament (ACL) were measured by biomechanical method. The tensile test was performed after the tendon was restored, and the load-prolongation curves were recorded. The maximum strain and maximum stress were calculated according to the previously measured initial length and cross-sectional area, and the total RNAs were extracted and reversed into cDNA. Real-time quantitative PCR was used to detect the common MMPs genes associated with tendon repair in the anteromedial and posterolateral bundles, and then fixed and paraffin embedded the two functional fiber bundles of ACL, the anterior medial bundle and the posterolateral bundle. Using the polyclonal antibody of MMP-2 and MMP-3 as the first antibody, the immunohistochemical experiment was carried out, the tissues of the anterior medial bundle and the posterolateral bundle were collected separately, the total protein was extracted from homogenate, and the expression of MMP-2mpMMP-3 in different bundles was analyzed by Western blotting. The product of gray scale and band area is the relative expression level of each protein, and the expression level of MMPs in different parts of anterior cruciate ligament is detected. Results:. 1. The structures and mechanical characteristics of different parts of anterior cruciate ligament (ACL) were measured by imaging and biomechanics, the maximum stress and strain of anterior internal bundle were larger than that of posterior external bundle. It is further demonstrated that the anterior tibial insertion of the anterior cruciate ligament has the potential to be the best site for clinical tendon reconstruction, which provides a theoretical basis for clinical application in the future. 2. The expression of MMPs in different parts of anterior cruciate ligament (ACL) was detected by RT-PCR and Western blot. The results showed that the expression of MMP-2 was higher in the anterior medial bundle, lower in the posterolateral bundle, and higher in the anteromedial bundle and posterolateral bundle. It is further demonstrated that the anteromedial bundle containing the anterior tibia has more remodeling potential than the posterolateral bundle. Conclusion:. The anterior medial bundle containing the front of the tibia is more important than the posterolateral bundle. The anterior end of the ACL tibia has a potential site for clinical tendon repair and remodeling.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R686.5

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