旋毛虫预感染对小鼠急性脊髓损伤炎症反应的抑制作用  

INHIBITORY EFFECT OF TRICHINELLA SPIRALIS PRE-INFECTION ON INFLAMMATORY RESPONSE IN MICE WITH ACUTE SPINAL CORD INJURY

作　　者：徐常艳 王小莉 万晨 杨浩宇 邱禹 许梦梦 吴宜修 常雪莲 袁圆 李徽徽 褚亮[3] 杨小迪 XU Chang-yan;WANG Xiao-li;WAN Chen;YANG Hao-yu;QIU Yu;XU Meng-meng;WU Yi-xiu;CHANG Xue-lian;YUAN Yuan;LI Hui-hui;CHU Liang;YANG Xiao-di(School of Basic Medicine,Bengbu Medical College,Bengbu 233030,Anhui,China;Key Laboratory of Infection and Immunity of Anhui Province,Bengbu 233030,Anhui,China;The Second Affiliated Hospital of Bengbu Medical College,Bengbu 233030,Anhui,China)

机构地区：[1]蚌埠医学院基础医学院,安徽蚌埠233030 [2]感染与免疫安徽省重点实验室,安徽蚌埠233030 [3]蚌埠医学院第二附属医院,安徽蚌埠233030

出　　处：《寄生虫与医学昆虫学报》2022年第1期1-8,共8页Acta Parasitologica et Medica Entomologica Sinica

基　　金：安徽省自然科学基金(No.2008085MH260);安徽省高校学科(专业)拔尖人才学术资助项目(No.gxbjZD15);安徽省教育厅高校优秀人才支持计划项目(No.gxyq2021188);安徽省教育厅重点自然科学研究项目(No.KJ2020A0566,No.KJ2020A0554,No.KJ2020A0572);蚌埠医学院自然科学高层次科技项目立项攀登计划项目(No.2020bypd007);蚌埠医学院“512人才培育计划”(No.by51201306);蚌埠医学院研究生科研创新计划项目(No.Byycx20006);蚌埠市-蚌埠医学院联合科技攻关项目(No.BYLK201818);省级大学生创新训练项目(No.S202010367108)。

摘　　要：为探讨旋毛虫预感染对小鼠急性脊髓损伤(spinal cord injury,SCI)的干预效果,选取40只C57BL/6雌性小鼠随机分为4组进行实验:A组(假手术组)、B组(旋毛虫预感染+假手术组)、C组(脊髓损伤组)、D组(旋毛虫预感染+脊髓损伤组)。A组小鼠只打开椎板,不损伤脊膜及脊髓;B组小鼠以旋毛虫肌幼虫(350条/只)灌胃后14 d打开椎板;C组小鼠进行SCI造模;D组小鼠在旋毛虫肌幼虫感染14 d后进行SCI造模。在术后第7天采用Basso小鼠评分(Basso mouse scale,BMS)观察不同组别小鼠后肢运动功能变化,其后收集小鼠脊髓组织和血清,HE染色法观察脊髓损伤处病理学变化及炎性细胞浸润情况;RT-PCR分析促炎因子IL-6、TNF-α和调节性细胞因子IL-10和TGF-β基因表达水平;ELISA检测血清IL-6、TNF-α、IL-10和TGF-β分泌水平。结果显示,术后第7天,4组小鼠行为学评分及单位面积的炎性细胞浸润数量差异均有统计学意义(P<0.05);C组小鼠行为学评分明显低于A组(P<0.05),且脊髓组织结构出现明显损伤,炎性细胞浸润显著增加(P<0.05);经旋毛虫预感染干预后,D组BMS评分较C组明显改善(P<0.05),且脊髓病理损伤显著缓解,炎性细胞浸润亦明显减少(P<0.05)。RT-PCR和ELISA结果显示:与C组相比,D组小鼠脊髓匀浆和血清中促炎细胞因子IL-6和TNF-α的表达均明显降低(P<0.05);抗炎细胞因子IL-10和TGF-β的表达均明显升高(P<0.05)。结果表明,旋毛虫预感染可抑制小鼠急性SCI后的炎症反应,减轻脊髓组织病理损伤,改善SCI小鼠肢体功能。To investigate the effects of Trichinella spiralis pre-infection on acute spinal cord injury(SCI)in mice,forty female C57BL/6 mice were randomly divided into 4 groups.Group A(sham operation group),group B(T.spiralis+sham group),group C(SCI group),group D(T.spiralis+SCI group).In group A,only the lamina was opened without damaging the spinal membrane and spinal cord.The lamina of mice in group B was opened 14 days after intragastrically fed with T.spiralis muscle larvae(350/mouse);Mice in group C were used for SCI modeling;Mice in group D were also used for SCI modeling 14 days after infection with T.spiralis muscle larvae.Basso Mouse Scale(BMS)score was used to observe the changes of hind limb motor function in different groups of mice on the 7th day after operation.Then the spinal cord tissue and serum of mice were collected and the pathological changes and inflammatory cell infiltration of spinal cord injury were observed by HE staining.The expression levels of pro-inflammatory cytokines IL-6,TNF-αand regulatory cytokines IL-10 and TGF-βwere analyzed by RT-PCR,and the levels of serum IL-6,TNF-α,IL-10 and TGF-βwere detected by ELISA.The results showed that the behavioral score and the number of inflammatory cell infiltration per unit area of the 4groups had statistical significance on the 7th day after operation(P<0.05);The behavioral score of mice in group C was significantly lower than that in group A(P<0.05),the spinal cord tissue structure showed obvious injury,and inflammatory cell infiltration was significantly increased(P<0.05);The BMS score of group D was significantly improved compared with group C after the intervention of T.spiralis pre-infection(P<0.05),spinal cord pathological injury was significantly alleviated,and inflammatory cell infiltration was significantly reduced(P<0.05).RT-PCR and ELISA results showed that the expression of pro-inflammatory cytokines IL-6 and TNF-αin spinal cord homogenate and serum of mice in group D decreased significantly compared with that in group C(P<0.05);The e

关 键 词：脊髓损伤 旋毛虫 炎症 Real-Time PCR ELISA 

分 类 号：R651.2[医药卫生—外科学]