RGMa通过Rho激酶调控EAE小鼠IL-27的表达和神经功能  

RGMa regulates IL-27 expression and neural function in EAE mice through Rho kinase

作　　者：牟春地 燕伟平 余刚[2] 秦新月[2] 冯金洲[2] MOU Chundi;YAN Weiping;YU Gang;QIN Xinyue;FENG Jinzhou(Department of Neurology,People′s Hospital of Qijiang District,Chongqing 401420,China;Department of Neurology,the First Affiliated Hospital of Chongqing Medical University,Chongqing, 400016,China;Department of Neurology,People′s Hospital of Guangrao County,Dongying,Shandong 257300,China)

机构地区：[1]重庆市綦江区人民医院神经内科,401420 [2]重庆医科大学附属第一医院神经内科,400016 [3]山东省广饶县人民医院神经内科,山东东营257300

出　　处：《重庆医学》2021年第20期3421-3425,共5页Chongqing medicine

基　　金：国家自然科学基金青年科学基金项目(81701191);重庆市教委科学技术研究项目(KJ1600210)。

摘　　要：目的探讨排斥性导向分子a(RGMa)对实验性自身免疫性脑脊髓炎(EAE)小鼠神经功能的影响,以及RGMa调控IL-27的相关分子机制。方法C57BL/6小鼠分为正常对照组,EAE模型组,RGMa RNAi组(rAd5-shRNA-RGMa重组腺病毒),空载体对照组(空载体rAd5-HK重组腺病毒),Rho激酶抑制剂组和PBS组,每组10只。除正常对照组外均构建EAE小鼠模型,每天行神经功能评分评价小鼠神经功能,造模当天记做免疫第0天,免疫12 d后对RGMa RNAi组、空载体对照组、Rho激酶抑制剂组及PBS组分别给予rAd5-shRNA-RGMa重组腺病毒[(2.51×10^(10))pfu/mL,6μL]、空载体rAd5-HK重组腺病毒[(2.51×10^(10))pfu/mL,6μL]、Rho激酶特异性抑制剂Y-27632(按体质量10 mg/kg)、等体积PBS干预,正常对照组及EAE模型组无其他干预。免疫18 d后行Western blot检测血清RGMa及Rho激酶的表达水平,ELISA检测血清IL-27的表达。结果Western blot结果显示RGMa RNAi组小鼠RGMa及Rho激酶表达较EAE模型组显著降低(0.07±0.01 vs.0.58±0.09,P<0.001;42.98±5.69 vs.66.74±8.35,P=0.027)。神经功能评分示RGMa RNAi组小鼠起病时间较EAE模型组及空载体对照组明显推迟,神经功能损伤明显减轻(P<0.05或P<0.01)。ELISA结果显示,与EAE模型组(1145.36±134.45)比较,RGMa RNAi组(658.46±83.41,P=0.004)及Rho激酶抑制剂组(804.45±133.90,P=0.019)IL-27表达水平显著降低,与PBS组(1060.73±104.54)比较,Rho激酶抑制剂组IL-27表达水平显著降低(P=0.022)。结论特异性抑制EAE小鼠RGMa可显著缓解其神经功能损伤,抑制IL-27的表达水平,这一作用可通过Rho激酶介导。Objective To investigate the effect of rejection guide molecule a(RGMa)on the neurological function of experimental autoimmune encephalomyelitis(EAE)mice,and the related molecular mechanism of RGMa regulating IL-27.Methods C57BL/6 mice were divided into the normal control group,the EAE model group,the RGMa RNAi group(rAd5-shRNA-RGMa recombinant adenovirus),empty vector control group(empty vector rAd5-HK recombinant adenovirus),the Rho kinase inhibitor group and the PBS group(n=10 per group).Except for the normal control group,EAE mouse models were constructed in the other groups.The neurological function scores were recorded every day to evaluate the neurological function of mice.The day of modeling was recorded as the 0th day of immunization.12 days after immunization,the RGMa RNAi group,the empty vector control group,the Rho kinase inhibitor group and the PBS group were given rAd5-shRNA-RGMa[(2.51×10^(10))pfu/mL,6μL],empty vector rAd5-HK[(2.51×10^(10))pfu/mL,6μL],Rho kinase specific inhibitor Y-27632(10 mg/kg)and equal volume of PBS intervention,respectively.Western blotwas performed to detect the expression levels of serum RGMa and Rho kinase at 18 days after immunization,and the expression of serum IL-27 was detected by ELISA.Results Western blot results showed that the expression leves of RGMa and Rho kinase in the RGMa RNAi group were significantly lower than that of the EAE model group(0.07±0.01 vs.0.58±0.09,P<0.001;42.98±5.69 vs.66.74±8.35,P=0.027).The neurological function score results showed that the onset of disease attack time of mice in the RGMa RNAi group was significantly delayed and the neurological damage was significantly alleviated when compared with the empty vector control group(P<0.05,P<0.01).ELISA results showed that,IL-27 expression level in the RGMa RNAi group(658.46±83.41,P=0.004)and the Rho kinase inhibitor group(804.45±133.90,P=0.022)was significantly reduced,when compared with the EAE model group(1145.36±134.45).The expression level of IL-27 in the Rho kinase inhibitor gro

关 键 词：多发性硬化 实验性自身免疫性脑脊髓炎 RGMa RHO激酶 白介素-27 

分 类 号：R744.51[医药卫生—神经病学与精神病学]