新鲜和冷冻人脐带来源间充质干细胞对大鼠骨关节炎的治疗作用比较  

Comparison of therapeutic effects of fresh and frozen human umbilical cord-derived mesenchymal stem cells on osteoarthritis in rats

作　　者：张斌斌 赵英杰[1] 杨雪枝 韩萍 常艳[1] 魏伟[1] ZHANG Binbin;ZHAO Yingjie;YANG Xuezhi;HAN Ping;CHANG Yan;WEI Wei(Institute of Clinical Pharmacology, Anhui Medical University,Key Lab of Anti-inflammatory and Immune Medicine, Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine, Hefei 230032, Anhui,China)

机构地区：[1]安徽医科大学临床药理研究所抗炎免疫药物教育部重点实验室抗炎免疫药物安徽省协同创新中心

出　　处：《中国临床药理学与治疗学》2019年第10期1092-1100,共9页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金资助项目(81573443、81673444);安徽省自然科学杰出青年基金(170808J10);2016年高校优秀青年人才支持计划重点项目(gxyq ZD2016043);安徽医科大学第四批“青年拔尖人才支持计划”

摘　　要：目的:比较新鲜和冷冻人脐带来源间充质干细胞对Ⅱ型胶原酶诱导的大鼠骨关节炎的治疗作用。方法:选用Sprague-Dawley (SD)大鼠,采用Ⅱ型胶原酶关节腔注射法建立大鼠骨关节炎模型,将大鼠随机分为正常组(Normal)、模型组(Model)、新鲜人脐带来源间充质干细胞组(Fresh hUC-MSC)、冷冻人脐带来源间充质干细胞组(Frozen hUC-MSC)和透明质酸组(HA),每组10只,于第一次注射Ⅱ型胶原酶后第7天给药。Fresh hUC-MSC和Frozen hUC-MSC组大鼠分别膝关节腔注射Fresh hUC-MSC和Frozen hUC-MSC 1×10^6/50 μL;HA组大鼠给予HA 50 μL;正常组和模型组大鼠给予等体积无菌生理盐水。给药后6周处死大鼠,观察各项指标。检测各组大鼠膝关节周长差值、关节软骨大体观察评分、膝关节X射线观察及评分、膝关节病理变化以及膝关节软骨基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)和基质金属蛋白酶抑制因子-1(tissue inhibitor of metalloproteinase-1,TIMP-1)的表达情况。结果:与模型组相比,Fresh hUC-MSC和Frozen hUC-MSC组关节周长差值和关节软骨大体评分均显著减小,差异均有统计学意义(P<0.01);Fresh hUC-MSC和Frozen hUC-MSC组X射线的凯尔格伦-劳伦斯分级评分以及番红固绿染色病理变化的Mankin's评分差异均有统计学意义(P<0.05)。同时,Fresh hUC-MSC和Frozen hUC-MSC组的MMP-13的表达显著降低,TIMP-1的表达显著升高,MMP-13和TIMP-1的比值显著降低,并且差异均具有统计学意义(P<0.01)。结论:Fresh hUC-MSC和Frozen hUC-MSC均能改善大鼠骨关节的损伤,两组治疗骨关节炎差异无统计学意义,为临床应用hUC-MSC治疗提供实验依据。AIM:To observe the therapeutic effect of fresh and frozen human umbilical cord-derived mesenchymal stem cells on type II collagenase-induced osteoarthritis in rats. METHODS:Sprague-Dawley (SD) rats with the model of osteoarthritis established by intra-articular injection of type II collagenase after one week of modeling, which were randomly divided into normal group (Normal), model group(Model), fresh human umbilical cord-derived mesenchymal stem cell group (Fresh hUC-MSC), and frozen human umbilical cord-derived mesenchymal stem cell group (Frozen hUC-MSC) and hyaluronic acid group (HA), with the number of 10 in each group. Rats in the Fresh hUC-MSC or Frozen hUC-MSC were injected into the knee joint cavity with 1×10^6/50 μL of fresh or frozen human umbilical cord-derived mesenchymal stem cells, respectively, in HA were given 50 μL of hyaluronic acid, and in Normal and Model group were given an equal volume of sterile physiological saline. The rats were sacrificed 6 weeks after the administration to observe various indexes.The knee circumference difference, articular cartilage gross observation score, knee X-ray, knee joint pathological changes and the expression of knee cartilage matrix metalloproteinase-13 (MMP-13) and matrix metalloproteinase inhibitor-1(TIMP-1) were detected in each group. RESULTS:Compared with the model group, the difference of joint circumference and the total score of articular cartilage in Fresh hUC-MSC and Frozen hUC-MSC group were significantly decreased, and the difference was statistically significant (P<0.01). The Kelgren-Lawrence grading scores of X-rays in Fresh hUC-MSC and Frozen hUC-MSC group and Mankin's scores in pathological changes of Safranin green staining were significantly different (P<0.05). At the same time, the expression of MMP-13 was significantly decreased in Fresh hUC-MSC and Frozen hUC-MSC group, the expression of TIMP-1 was significantly increased, the ratio of MMP-13 and TIMP-1 was significantly decreased, and the difference was statistically significant(P<0

关 键 词：新鲜人脐带来源间充质干细胞 冷冻人脐带来源间充质干细胞 骨关节炎 

分 类 号：R965.2[医药卫生—药理学] R683[医药卫生—药学]