脂肪来源间充质干细胞调控变应性鼻炎小鼠T细胞免疫状态的研究  

Effect of adipose-derived mesenchymal stem ceils （ADSC） on the T cell immune status of allergicrhinitis mouse model

作　　者：李冠雪[1] 刘艳慧[2] 申聪香[1] 文忠[1] 张沈华[1] 杨柯柯[1] 

机构地区：[1]南方医科大学珠江医院儿科小儿耳鼻咽喉组,广州510282 [2]新疆医科大学附属二院耳鼻咽喉科,乌鲁木齐830285

出　　处：《中华耳鼻咽喉头颈外科杂志》2016年第1期50-56,共7页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的 探讨脂肪来源间充质干细胞（adipose—derivedstemceils，ADSC）对变应性鼻炎（allergicrhinitis，AR）小鼠辅助性T细胞（Th）、调节性T细胞（Treg）的调节作用及其机制。方法取Balb／c小鼠60只，采用随机数字表法随机分为6组（分组表示为：致敏／激发／处理），分别为实验组1（OVA／OVA／高剂量ADSC）、实验组2（OVA／OVA／低剂量ADSC）、实验组3（0VA／OVA／磷酸盐缓冲液PBS）、实验组4（OVA／OVA／0）、对照组1（PBS／PBS／0）和对照组2（O／O／0）。常规分离提取小鼠ADSC，卵清蛋白（ovalbumin，OVA）联合氢氧化铝构建AR小鼠模型，尾静脉注射CM-Dil标记的ADSC（高剂量组：3×10^6；低剂量组：1×10^6）连续3d，48h后处死，酶联免疫吸附试验（enzymelinkedimmunosorbentassay，ELISA）检测血清中自细胞介素（interleukin，IL）4、IL-6、IL-10和叮干扰素（Y interferon，IFN-y）的水平，反转录聚合酶链反应（reversetranscriptasepolymerasechainreaction，RT—PCR）检测脾脏中上述各细胞因子mRNA的表达，荧光显微镜下观察ADSC在AR模型鼠鼻黏膜中的迁移分布，鼻黏膜HE染色观察嗜酸粒细胞浸润情况。以SPSS13．0软件进行统计学处理。结果成功分离培养并鉴定小鼠ADSC。AR模型小鼠症状学评分与对照组比较差异有统计学意义（P〈0．01）。实验组1与实验组4相比，血清IL4、IL-6水平下调[组1：（17．95±7．78）、（27．51±5．93）pg／ml；组4：（56．82±9．12）、（70．034-7．22）pg／m1]，IFN-1、IL-10水平上调[组1：（367．74±、13．79）、（417．10±72．40）p∥ml；组4：（199．464-11．25）、（122．50±15．57）pgZml]，差异均有统计学意义（P值均〈0．01）。实验组1与实验组2相比，血清IL-4、IL．6水平下调[组1：（17．95±7．78）、（27．51±5．93）pg／ml；组2：（41．57±12．27）、（56．21±9．23）pg／m1]，IFN-1、IL-10水平上调[组1�Objective To investigate the regulation of adipose-derived rnesenchyrnal stern cells （ADSC） on helper T ceils and regulatory T cells in allergic rhinitis （AR） mouse model and the underlying mechanisms. Methods Using random number table, 60 Balb/c mice were divided into 6 groups （represented by： sensitized/challenged/treated ）, they were the experimental group 1 （ OVA/OVA/high dose ADSC）, the experimental group 2（ OVA/OVA/low dose ADSC）, the experimental group 3 （OVA/ OVA/PBS） , the experimental group 4 （ OVA/OVA/0）, the control group 1 （PBS/PBS/0） and the control group 2 （0/0/0）. The mouse ADSC were isolated and cultured through conventional method, and AR mouse model was built with OVA and aluminum. The mice were injected with high （ 3 × 10^6 ） , low （ 1 × 10^6 ） ADSC respectively labeled by CM-I）il for 3 consecutive days via tail-vein injection and sacrificed 48 hours later. Finally, levels of IL-4, IL-6, IL-10 and IFN -y in serum were examined by ELISA; expressions of the four cytokines in spleen were examined by q RT-PCR; migration of ADSC to mouse model nasal mucosa were observed through fluorescence microscope; eosinophil infiltration were observed by the nasal HE staining. Results Mouse ADSC was isolated, cultured and identified successfully. There was significant difference in symptom scores of AR models （ compared with 0/0/0 group, P 〈 0.01 ）. The IL-4 and IL-6 levels of OVA/ OVA/high ADSC group were significantly lower than OVA/OVA/0 group （ group 1 ： （ 17.95 ± 7.78 ）, （27.51 ± 5.93 ） pg/ml; group 4： （56. 82 ± 9.12）, （70. 03±7. 22） pg/ml）, the IFN-~/ and IL-10 levels increased significantly （ group 1 ： （367.74 ± 13.79 ）, （417.10 ± 72. 40） pg/ml ; group 4 ： （ 199.46± 11.25 ）, （ 122. 50 -＋ 15.57）pg/ml） in serum. These differences were statistically significant（P 〈0. 01 ）. Compared with OVA/OVA/low ADSC group, the IL-4 and IL-6 levels decreased significantly （ group 1 ： （ 17.95 ±7.7

关 键 词：鼻炎 变应性 脂肪来源间充质干细胞 Th1/Th2细胞免疫平衡 调节性T细胞 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]