CTLA4-Ig腹腔注射对大鼠实验性自身免疫性重症肌无力的治疗作用及其机制  

Therapeutic effect of intraperitoneal injection of CTLA4-Ig on EAMG in rats and its mechanism

作　　者：焦子洋 梁培鑫 薛占霞[1] 郑立卿[1] 董晓华[1] 吴苗苗[1] 武海霞[1] 侯勇[1] JIAO Ziyang;LIANG Peixin;XUE Zhanxia;ZHENG Liqing;DONG Xiaohua;WU Miaomiao;WU Haixia;HOU Yong(Department of Pharmacy,Hebei North University,Hebei Key Laboratory of Neuropharmacology,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院药学院,河北省神经药理学重点实验室,河北张家口075000

出　　处：《山东医药》2022年第30期14-19,共6页Shandong Medical Journal

基　　金：省属高校基本科研业务费项目(JYT2020022)。

摘　　要：目的观察细胞毒性T淋巴细胞抗原4免疫球蛋白(CTLA4-Ig)腹腔注射对大鼠实验性自身免疫性重症肌无力(EAMG)的治疗作用,并探讨其可能作用机制。方法取45只雌性Lewis大鼠,其中35只大鼠采用AchRα97-116肽免疫法构建EAMG模型(第1、30、60天皮下注射50μg AChRα97-116肽),另10只大鼠(对照组)同期给予等量生理盐水。第74天时将造模成功的EAMG模型大鼠随机分为CTLA4-Ig组、EAMG组及地塞米松组,每组各10只,CTLA4-Ig组大鼠用2 mg/kg CTLA4-Ig腹腔注射,地塞米松组用1 mg/kg地塞米松腹腔注射,EAMG组及对照组大鼠腹腔注射相同体积PBS,1天1次至第130天。分别于第74、102、130天对各组大鼠进行称重,观察肌力变化,采集尾静脉血,第130天同时采集大鼠后肢肌肉组织和胸腺组织,ELISA法检测血清AChR IgG、AChR IgG 2b及后肢肌肉组织AChR含量,RT-PCR法检测胸腺组织凋亡相关基因Fas、FasL、B细胞淋巴瘤-2(Bcl-2)、Bax、半胱氨酸天冬氨酸特异蛋白酶3(Caspases-3)、Caspases-8和Caspases-9的mRNA,紫外分光法检测大鼠胸腺组织Caspases-3、Caspases-8和Caspases-9活性。结果与对照组比较,EMAG组大鼠Lennon评分高、肌肉AChR含量少、血清AChR IgG、AChR IgG2b水平高(P均<0.05);与EAMG组比较,第102、130天CTLA4-Ig组及地塞米松组大鼠体质量升高、Lennon评分低,血清AChR IgG、AChR IgG 2b水平降低,后肢肌肉组织AChR含量增加(P均<0.05)。与对照组比较,EAMG组大鼠胸腺组织Fas mRNA、Bcl-2 mRNA相对表达量高,胸腺组织Caspases-3 mRNA、Caspases-8 mRNA和Caspases-9 mRNA相对表达量降低,胸腺组织Caspases-3、Caspases-8和Caspases-9活性低(P均<0.05);与EAMG组比较,CTLA4-Ig组和地塞米松组FasL mRNA、Bax mRNA、Caspases-3 mRNA、Caspases-8 mRNA和Caspases-9 mRNA相对表达量高,Bcl-2 mRNA相对表达量低,Caspases-3、Caspases-8和Caspases-9活性高(P均<0.05)。结论CTLA4-Ig腹腔注射可改善EAMG大鼠的临床症状。CTLA4-Ig可能通过上调胸腺组Objective To evaluate the therapeutic effect of intraperitoneal injection of cytotoxic T lymphocyte antigen 4 immunoglobulin(CTLA4-Ig)on experimental autoimmune myasthenia gravis(EAMG)in rats,and to investigate its underlying mechanism.Methods Forty-five female Lewis rats were selected,among which,35 rats were immunized with AchRα97-116 peptide to establish EAMG models(50μg AChRα97-116 peptide was injected subcutaneously on the 1st,30th and 60th days),and the other 10 rats(control group)were given the same amount of normal saline at the same time.On the 74th day,the successfully modeled EAMG rats were randomly divided into the CTLA4-Ig group,EAMG group and dexamethasone group,with 10 rats in each group.Rats in the CTLA4-Ig group were intraperitoneally injected with 2 mg/kg CTLA4-Ig,dexamethasone group with 1 mg/kg dexamethasone,and EAMG group and the control group with the same volume of PBS,once a day to the 130th day.On the 74th,102nd and 130th days,the rats in each group were weighed,the changes of muscle strength were observed,and the tail vein blood was collected.On the 130th day,the muscle tissue and thymus tissue of hind limbs of rats were collected at the same time.Serum levels of AChR IgG and AChR IgG 2b and the content of AChR in hind limbs were detected by ELISA.The mRNA expression levels of Fas,FasL,B-cell lymphoma-2(Bcl-2),B-cell lymphoma-2 associated X protein(Bax),cysteinyl aspartate specific protease-3(Caspases-3),Caspase-8 and Caspase-9 in thymus were measured by RT-PCR.The activities of Caspases-3,Caspase-8 and Caspase-9 in thymus were measured by ultraviolet spectrophotometry.Results Compared with the control group,the rats in the model group had higher Lennon score,less AChR content in muscle and higher levels of AChR IgG and AChR IgG2b in serum(all P<0.05).Compared with EAMG group,the rats in the CTLA4-Ig group and dexamethasone group had higher body weight,lower Lennon score,lower levels of serum AChR IgG and AChR IgG2b,and higher AChR content in hind limb muscles on the 102nd and 130th d

关 键 词：细胞毒性T淋巴细胞抗原4 细胞毒性T淋巴细胞抗原4免疫球蛋白 凋亡相关基因 细胞凋亡 重症肌无力 实验性自身免疫性重症肌无力 

分 类 号：R392[医药卫生—免疫学]