Toll样受体3预处理增强羊膜间充质干细胞对大鼠创伤性脑损伤的治疗作用  

Toll-like receptor 3 preconditioning enhances the therapeutic efficacy of amniotic mesenchymal stem cells on traumatic brain injury in rat

作　　者：闫中杰[1] 赵宗茂[1] 李凤菊 刘力强[1] 杨亮[1] 李琛[1] 石云鹏 卢圣奎[1] Yan Zhongjie;Zhao Zongmao;Li Fengju;Liu Liqiang;Yang Liang;Li Chen;Shi Yunpeng;Lu Shengkui(Department of Neurosurgery,the Second Hospital of HebeiMedical University,Shijiazhuang 050003,China;Department of Radiology,Hebei Armed Police Corps Hospital,Shijiazhuang 050050,China)

机构地区：[1]河北医科大学第二医院神经外科,石家庄050003 [2]武警河北总队医院放射科,石家庄050050

出　　处：《中华实验外科杂志》2020年第8期1472-1476,共5页Chinese Journal of Experimental Surgery

基　　金：2017年河北省政府资助临床医学优秀人才培养和基础课题研究项目。

摘　　要：目的探讨Toll样受体3(TLR3)通路激活对羊膜间充质干细胞移植治疗大鼠创伤性脑损伤的作用及其机制。方法TLR3激活可使间充质干细胞(MSC)进入抑制免疫状态(M2)。应用聚肌胞苷酸[poly(I∶C),TLR3的配体]预处理羊膜间充质干细胞(AMSC)。采用SD大鼠(购自河北医科大学实验动物中心),应用随机数法随机分为3组,生理盐水组、AMSC组、预处理组,将细胞移植到创伤性脑损伤(TBI)大鼠损伤周围区域,应用改良神经功能损伤评分(mNSS),评估poly(I∶C)预处理对AMSC治疗大鼠TBI是否存在增强效应;应用免疫组织荧光法检测损伤局部的炎性反应强度,以及局部巨噬细胞/小胶质细胞的极化状态,并应用酶联免疫吸附试验(ELISA)法检测损伤局部鼠源性炎性因子白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、IL-10、转化生长因子-β(TGF-β)的表达水平,分析TLR3激活的AMSC对TBI大鼠治疗作用的机制。组间比较采取重复测量方差分析和LSD检验。结果将poly(I∶C)预处理后的AMSC移植入TBI大鼠体内后,移植后不同时间点mNSS神经功能评分显示,预处理组移植术后自第14天起均优于AMSC组:14 d(4.28±0.38比5.35±0.41,F=168.668,P<0.05)、21 d(4.08±0.35比4.46±0.39,F=192.316,P<0.05)、28 d(3.56±0.29比4.06±0.34,F=283.572,P<0.05);与AMSC组比较,预处理组能进一步降低损伤局部的炎性反应,促进损伤区域巨噬细胞/小胶质细胞极化进入抗炎的M2状态;移植后损伤局部鼠源性炎性因子检测结果显示,移植后1 d炎症急性期,预处理组促炎因子低于AMSC组[IL-1β,(40.6±4.1)ng/L比(45.1±4.8)ng/L,F=47.147,P<0.05、TNF-α,(150.2±18.0)ng/L比(180.0±20.6)ng/L,F=53.028,P<0.05],抗炎因子高于AMSC组[IL-10,(35.3±2.8)ng/L比(28.7±1.9)ng/L,F=96.934,P<0.05、TGF-β,(20.9±2.6)ng/L比(17.6±1.4)ng/L,F=47.305,P<0.05]。结论poly(I∶C)预处理可增强AMSC对TBI大鼠的治疗作用,其机制可能是通过诱导局部免疫细胞极化,调节免疫微�Objective To investigate the effect of Toll-like receptor 3(TLR3)pathway activation on the treatment of traumatic brain injury in rats with amnion mesenchymal stem cell transplantation.Methods TLR3 activation can cause mesenchymal stem cells(MSC)to enter a suppressed immune state(M2).Polyinosinic acid[poly(I∶C),ligand of TLR3]was used to pretreat amniotic mesenchymal stem cells(AMSC).SD rats were used,purchased from the Experimental Animal Center of Hebei Medical University,and randomly divided into 3 groups using random number method,physiological saline group,AMSC group,and pretreatment group.Cells were transplanted to the area around the traumatic brain injury(TBI)rat injury.The modified neurological severity scores(mNSS)neurological function score was used to evaluate poly(I∶C)Whether pretreatment has an enhanced effect on the treatment of TBI by AMSC in rats;immunohistofluorescence method was used to detect the intensity of the inflammatory response in the injured area,and the polarization state of local macrophages/microglia,and the enzyme linked immunosorbent assay(ELISA)method was used to detect the injured rats The expression levels of inflammatory factors interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-10 and transforming growth factor-β(TGF-β)were analyzed to analyze the mechanism of AMSC activated by TLR3 on the treatment of TBI rats.Comparison between groups adopts repeated measures analysis of variance and LSD test.Results After poly(I∶C)pretreatment AMSC was transplanted into TBI rats,the mNSS neurological function scores at different time points after transplantation showed that the preconditioning group was better than the AMSC group from the 14th day after transplantation:14 d(4.28±0.38 vs.5.35±0.41,F=168.668,P<0.05),21 d(4.08±0.35 vs.4.46±0.39,F=192.316,P<0.05),28 d(3.56±0.29 vs.4.06±0.34,F=283.572,P<0.05);compared with AMSC group,pretreatment group It can further reduce the inflammatory response in the injured area,and promote the polarization of macrophages/microglia in

关 键 词：创伤性脑损伤 羊膜间充质干细胞 免疫调节 TOLL样受体3 炎症 

分 类 号：R651.15[医药卫生—外科学] R-332[医药卫生—临床医学]