绿色荧光蛋白示踪检测骨髓间充质干细胞移植在CIA大鼠体内的迁移、定植和分布

发布时间：2018-05-16 11:04

本文选题：胶原诱导关节炎 + 绿色荧光蛋白　；参考：《山西医科大学》2015年硕士论文

【摘要】：目的:GFP标记的BM-MSCs通过体外培养、扩增后移植到CIA大鼠体内,通过光学成像联合免疫组化的方法检测其在CIA大鼠免疫器官以及关节部位的迁移、定植和分布情况,探讨其修复损伤的可能机制。方法:1.对GFP标记的MSCs进行体外培养、扩增及鉴定。2.将II型胶原与完全弗氏佐剂的混合制剂间隔14天两次免疫Wistar大鼠,从大鼠症状(大体观、关节炎指数、后肢肿胀程度)、影像学、病理学方面进行评估,建立CIA大鼠模型。随机分为早期干预组(n=20)、晚期干预组(n=20),同时设置CIA早期对照组(n=12)、晚期对照组(n=12)和正常对照组(n=12)。3.尾静脉途径移植GFP-MSCs,小动物活体成像技术动态观察移植MSCs在大鼠体内的迁移过程。4.于3、11、30和42天将各组大鼠的脾脏、胸腺、淋巴结和关节组织作成病理切片,通过免疫组化的方法检测移植细胞在免疫器官和炎症关节部位的迁移和分布。结果:1.(1)早、晚期干预组关节炎指数、关节肿胀程度较CIA对照组明显下降(P0.05);(2)早期干预组与晚期干预组比较,关节炎指数与关节肿胀程度降低,差异有统计学意义(P0.05)。2.小动物活体成像技术未能动态监测到MSCs移植在CIA大鼠体内的迁移过程,动物自身荧光干扰明显。3.抗GFP免疫组化法在干预组大鼠免疫器官和关节部位成功检测到GFP阳性结果,并可持续存在至少42天。结论:1.MSCs经尾静脉途径移植后,可迁移至CIA大鼠脾脏、淋巴结和胸腺和关节炎症部位,并可较长期(42天)定植于该组织。2.GFP不适用活体示踪检测MSCs在大鼠免疫器官及关节部位的迁移。3.MSCs干预对CIA大鼠有效,早期干预疗效优于晚期干预。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22;R457.7

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