Blau综合征动物模型的建立  

Establishment of animal model of Blau syndrome

作　　者：宋昊昕 杨熙玥 叶菜英[1] 朱蕾[1] SONG Hao-xin;YANG Xi-yue;YE Cai-ying;ZHU Lei(Dept of Pharmacology,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences,and School of Basic Medicine,Peking Union Medical College,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院药理系,北京100005

出　　处：《中国药理学通报》2023年第6期1195-1199,共5页Chinese Pharmacological Bulletin

基　　金：中国医学科学院医学与健康科技创新工程(No 2021-1-I2M-005)。

摘　　要：目的建立稳定可靠的Blau综合征(Blau syndrome,BS)体内动物模型。方法C57BL/6J小鼠腹腔注射胞壁酰二肽(muramyl dipeptide,MDP)或L18-MDP建立BS系统性炎症模型,同时设置阳性药依那西普(etanercept,ETN)考察模型对药物的响应以评价其有效性;SD大鼠玻璃体腔内注射MDP建立BS相关的葡萄膜炎模型;ELISA法检测血清中TNF-α、IL-6和IL-8水平;HE染色法检测眼球病理学变化;免疫组织化学染色法检测玻璃体内p65、p-ERK、p-p38、p-JNK、TNF-α、IL-6和IL-8的表达情况。结果小鼠血清中TNF-α水平在腹腔注射MDP后出现上升(P<0.05),在注射L18-MDP后明显升高(P<0.01),同时血清中IL-6和IL-8水平亦被L18-MDP明显诱导(P<0.01)。ETN对L18-MDP诱导的上述细胞因子水平的升高均有明显抑制作用(P<0.05,P<0.01)。大鼠玻璃体腔内注射MDP后,视网膜及玻璃体中有大量炎性细胞浸润,视网膜损伤严重,眼球组织中p65、p-ERK、p-p38、p-JNK、TNF-α、IL-6和IL-8的染色明显增强。结论C57BL/6J小鼠腹腔注射L18-MDP可成功构建系统性BS动物模型,SD大鼠玻璃体腔内注射MDP可建立良好的BS相关葡萄膜炎动物模型,这为探索BS发病机制以及开展药效评价提供了操作简便、容易重复的较为理想的动物模型。Aim To establish stable and reliable animal models of Blau syndrome(BS)in vivo.Methods C57BL/6J mice were intraperitoneally injected with muramyl dipeptide(MDP)or L18-MDP to induce systemic inflammatory model of BS.Meanwhile,positive drug etanercept(ETN)was set to investigate the response of the model to evaluate effectiveness.SD rats were intravitreally injected with MDP to establish BS-associated uveitis model.Serum levels of TNF-α,IL-6 and IL-8 were detected by enzyme linked immunosorbent assay(ELISA).Histopathological changes of rat eyeballs were detected by HE staining and the expressions of p65,p-ERK,p-p38,p-JNK,TNF-α,IL-6 and IL-8 in vitreous were determined by immunohistochemistry(IHC)staining.Results The serum level of TNF-αin mice increased after intraperitoneal injection of MDP(P<0.05),and increased significantly after L18-MDP injection(P<0.01).Meanwhile,the levels of IL-6 and IL-8 were also markedly induced by L18-MDP(P<0.01,P<0.01).ETN treatment evidently inhibited the increased levels of these above cytokines induced by L18-MDP(P<0.05,P<0.01).After the intravitreous injection of MDP in SD rats,there were numerous inflammatory cells infiltrated in retina and vitreous,and the retina was seriously damaged.The staining levels of p65,p-ERK,p-p38,p-JNK,TNF-α,IL-6 and IL-8 in eyeball tissues were significantly enhanced.Conclusions The systemic animal model of BS can be successfully established by intraperitoneal injection of L18-MDP in C57BL/6J mice,and the good BS-related uveitis can be induced by intravitreous injection of MDP in SD rats,which provides the simple,convenient,repeatable and ideal animal models for exploring the pathogenesis of BS and evaluating the efficacy of drugs.

关 键 词：Blau综合征 胞壁酰二肽 动物模型 系统性炎症 葡萄膜炎 细胞因子 

分 类 号：R-332[医药卫生] R363-332R392.12R773.9R977.6