本文選題：膠原誘導(dǎo)關(guān)節(jié)炎 + 綠色熒光蛋白��； 參考：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:GFP標(biāo)記的BM-MSCs通過體外培養(yǎng)、擴(kuò)增后移植到CIA大鼠體內(nèi),通過光學(xué)成像聯(lián)合免疫組化的方法檢測(cè)其在CIA大鼠免疫器官以及關(guān)節(jié)部位的遷移、定植和分布情況,探討其修復(fù)損傷的可能機(jī)制。方法:1.對(duì)GFP標(biāo)記的MSCs進(jìn)行體外培養(yǎng)、擴(kuò)增及鑒定。2.將II型膠原與完全弗氏佐劑的混合制劑間隔14天兩次免疫Wistar大鼠,從大鼠癥狀(大體觀、關(guān)節(jié)炎指數(shù)、后肢腫脹程度)、影像學(xué)、病理學(xué)方面進(jìn)行評(píng)估,建立CIA大鼠模型。隨機(jī)分為早期干預(yù)組(n=20)、晚期干預(yù)組(n=20),同時(shí)設(shè)置CIA早期對(duì)照組(n=12)、晚期對(duì)照組(n=12)和正常對(duì)照組(n=12)。3.尾靜脈途徑移植GFP-MSCs,小動(dòng)物活體成像技術(shù)動(dòng)態(tài)觀察移植MSCs在大鼠體內(nèi)的遷移過程。4.于3、11、30和42天將各組大鼠的脾臟、胸腺、淋巴結(jié)和關(guān)節(jié)組織作成病理切片,通過免疫組化的方法檢測(cè)移植細(xì)胞在免疫器官和炎癥關(guān)節(jié)部位的遷移和分布。結(jié)果:1.(1)早、晚期干預(yù)組關(guān)節(jié)炎指數(shù)、關(guān)節(jié)腫脹程度較CIA對(duì)照組明顯下降(P0.05);(2)早期干預(yù)組與晚期干預(yù)組比較,關(guān)節(jié)炎指數(shù)與關(guān)節(jié)腫脹程度降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。2.小動(dòng)物活體成像技術(shù)未能動(dòng)態(tài)監(jiān)測(cè)到MSCs移植在CIA大鼠體內(nèi)的遷移過程,動(dòng)物自身熒光干擾明顯。3.抗GFP免疫組化法在干預(yù)組大鼠免疫器官和關(guān)節(jié)部位成功檢測(cè)到GFP陽性結(jié)果,并可持續(xù)存在至少42天。結(jié)論:1.MSCs經(jīng)尾靜脈途徑移植后,可遷移至CIA大鼠脾臟、淋巴結(jié)和胸腺和關(guān)節(jié)炎癥部位,并可較長期(42天)定植于該組織。2.GFP不適用活體示蹤檢測(cè)MSCs在大鼠免疫器官及關(guān)節(jié)部位的遷移。3.MSCs干預(yù)對(duì)CIA大鼠有效,早期干預(yù)療效優(yōu)于晚期干預(yù)。
[Abstract]:Objective to investigate the migration, colonization and distribution of BM-MSCs labeled with GFP in the immune organs and joints of CIA rats by means of optical imaging combined with immunohistochemistry. To explore the possible mechanism of its repair injury. Method 1: 1. GFP labeled MSCs was cultured, amplified and identified in vitro. Wistar rats were immunized twice with collagen II and complete Freund adjuvant mixture for 14 days. The rat model of CIA was established by evaluating the symptoms (gross view, arthritis index, swelling degree of hind limbs, imaging and pathology). They were randomly divided into two groups: early intervention group, late intervention group, CIA early control group and late control group. GFP-MSCs was transplanted through the caudal vein and the migration process of transplanted MSCs in rats was observed dynamically by small animal in vivo imaging. The spleen, thymus, lymph nodes and joints of each group were made into pathological sections at 30 and 42 days after operation. The migration and distribution of transplanted cells in immune organs and inflammatory joints were detected by immunohistochemical method. Results (1) the arthritis index and joint swelling degree in the early and late intervention group were significantly lower than those in the CIA control group (P < 0.01). The arthritis index and the joint swelling degree in the early intervention group were significantly lower than those in the late intervention group, and the difference was statistically significant. In vivo imaging of small animals could not dynamically monitor the migration process of MSCs transplantation in CIA rats. The positive results of GFP were detected successfully in the immune organs and joints of the rats in the intervention group by immunohistochemical method of anti GFP, and the results were sustained for at least 42 days. Conclusion: 1. MSCs can migrate to spleen, lymph node, thymus and arthritis of CIA rats after transplantation via caudal vein. GFP could not be used to detect the migration of MSCs in the immune organs and joints of rats. 3. MSCs intervention was effective in CIA rats, and the early intervention effect was better than the late intervention.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R593.22;R457.7

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本文編號(hào)：1896581