Article(id=1208795421661590166, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208795418612339683, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2021.10.01, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1603555200000, receivedDateStr=2020-10-25, revisedDate=1624118400000, revisedDateStr=2021-06-20, acceptedDate=null, acceptedDateStr=null, onlineDate=1766128886857, onlineDateStr=2025-12-19, pubDate=1635350400000, pubDateStr=2021-10-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766128886857, onlineIssueDateStr=2025-12-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766128886857, creator=13701087609, updateTime=1766128886857, updator=13701087609, issue=Issue{id=1208795418612339683, tenantId=1146029695717560320, journalId=1189873630562394117, year='2021', volume='46', issue='10', pageStart='955', pageEnd='1060', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1766128886129, creator=13701087609, updateTime=1766128956061, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1208795711982924071, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208795418612339683, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1208795711982924072, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208795418612339683, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=955, endPage=960, ext={EN=ArticleExt(id=1208795421984551578, articleId=1208795421661590166, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=OPN/IGF1 promotes the corticospinal tract regeneration by increasing neuron IGFR expression, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objectives To study the role and associated potential molecular mechanism of osteopontin/insulin-like growth factor 1 (OPN/IGF1) in the corticospinal tract (CST) regeneration. Methods A total of 28 adult C57BL/6 mice (6-8 weeks old) were randomized into the experimental group (injection of AAV-OPN/IGF1, n=14) and control group (injection of AAV-PLAP, n=14) after training of single pellet retrieval. After received the unilateral pyramidotomy surgery, animals received AAV-OPN/IGF1 or AAV-PLAP at the injury site. Further, we employed the movement behaviour test to evaluate spinal motor recovery at 2-week intervals till 12-week after surgery. The protein kinase Cγ (PKCγ) staining in the dorsal funiculus of the cervical spinal cord indicated surgical efficiency. Lastly, we examined the axon sprouting and expression levels of ribosomal protein S6 kinase (pS6)and insulin-like growth factor receptor (IGFR) in neurons using immunohistochemical staining to study the potential mechanism of OPN/IGF1 to promote axon prouting. Results After evaluating PKCγ staining to indicate surgery effectiveness, a total of ten mice were in the experimental group and eight in the control group. In the mouse unilateral pyramidotomy model, OPN/IGF1 has not much effect on the recovery of motor function compared with the control group. After the overexpression of OPN/IGF1 on cortical neurons, sprouting of the spared CST was increased by the elevated expression of pS6 and IGFR, and the difference was statistically significant (P<0.05). Conclusions OPN/IGF1 promotes the sprouting of the CST by increasing the expression of neuronal IGFR and pS6.

, correspAuthors=Fang Cui, authorNote=null, correspAuthorsNote=
*E-mail:
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目的 研究骨桥蛋白(OPN)/胰岛素样生长因子1(IGF1)在小鼠皮质脊髓束(CST)再生中的作用及相关分子机制。方法 6~8周龄C57BL/6小鼠28只,随机分为实验组与对照组,每组14只。建立小鼠单侧锥体束横断模型,实验组在损伤对侧皮质注射腺相关病毒(AAV)-OPN/IGF1,对照组在损伤对侧皮质注射AAV-胎盘碱性磷酸酶(PLAP)。术后每2周采用行为学方法评估小鼠运动功能的恢复情况。术后12周处死小鼠,采用脊髓蛋白激酶Cγ(PKCγ)免疫组化染色判断皮质脊髓束横断情况;采用红色荧光蛋白(RFP)免疫组化染色分析轴突发芽的情况;采用免疫荧光染色检测神经元OPN和IGF1相关信号通路中核糖体蛋白S6激酶(pS6)和胰岛素样生长因子受体(IGFR)表达情况,研究OPN/IGF1促进轴突发芽的分子机制。结果 通过采用PKCγ免疫组化染色判断皮质脊髓束残余情况,最终实验组入组小鼠10只,对照组入组小鼠8只。实验组与对照组相比,OPN/IGF1对小鼠运动功能恢复无明显作用;皮质神经元高表达OPN/IGF1以后,损伤对侧皮质脊髓束发芽增多,神经元pS6和IGFR表达均明显高于对照组,差异有统计学意义(P<0.05)。结论 OPN/IGF1可通过增加神经元IGFR和pS6表达而促进CST发芽。

, correspAuthors=崔芳, authorNote=null, correspAuthorsNote=
崔芳,E-mail:
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张倩,博士研究生,主要从事脑血管病、神经系统变性病、周围神经病的诊疗研究

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张倩,博士研究生,主要从事脑血管病、神经系统变性病、周围神经病的诊疗研究

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BL. 基线;AAV. 腺相关病毒;PLAP. 胎盘碱性磷酸酶;IGF1. 胰岛素样生长因子1;A. 不规则步行梯测试[实验组(n=9),对照组(n=8)];B. 糖丸抓取测试[实验组(n=6),对照组(n=6)];C. 握食意大利面测试[实验组(n=9),对照组(n=7)]

, figureFileSmall=KUDqk3AFE/bOesus3prczw==, figureFileBig=AbA7lTQgGJ8IKULtUeNU3Q==, tableContent=null), ArticleFig(id=1208795432063464285, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208795421661590166, language=EN, label=Fig. 2, caption=Axon sprouted to the contralateral side in mice after unilateral pyramidotomy, figureFileSmall=nwJ9Zo5HKzcz56gcUMV7ng==, figureFileBig=BA9pQ1acU2AJuifV9xH7Ww==, tableContent=null), ArticleFig(id=1208795432176710500, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208795421661590166, language=CN, label=图2, caption=单侧锥体束横断术后小鼠未损伤侧轴突向对侧发芽情况

AAV. 腺相关病毒;PLAP. 胎盘碱性磷酸酶;IGF1. 胰岛素样生长因子1;A. 颈椎(C6)RPF免疫组化染色(显示未损伤侧轴突向损伤侧发芽情况,bar=500 μm);B. 轴突数量统计(通过中央导水管画一水平线,Mid、Z1、Z2是3条把灰质分为3等份的垂直线,统计通过三条垂直线的轴突数量);C. 实验组(n=8)和对照组(n=5)分别通过Mid、Z1、Z2三条垂直线的轴突数量相对于各自锥体交叉前标记轴突数量的比值的统计学分析,与对照组比较,(1)P<0.05,(2)P<0.01。

, figureFileSmall=nwJ9Zo5HKzcz56gcUMV7ng==, figureFileBig=BA9pQ1acU2AJuifV9xH7Ww==, tableContent=null), ArticleFig(id=1208795432277373802, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208795421661590166, language=EN, label=Fig. 3, caption=Expressions of neuronal pS6 and IGFR in mice after unilateral pyramidotomy, figureFileSmall=SP6KNLrKN2nxxjyAawpQgQ==, figureFileBig=E3XZ6KktnAsNorFB9DLXZg==, tableContent=null), ArticleFig(id=1208795432399008626, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208795421661590166, language=CN, label=图3, caption=单侧锥体束横断术后小鼠神经元pS6(A)及IGFR(B)的表达(n=6,bar=20 μm)

pS6. 核糖体蛋白S6激酶;IGFR. 胰岛素样生长因子受体;AAV. 腺相关病毒;PLAP. 胎盘碱性磷酸酶;IGF1. 胰岛素样生长因子1;与对照组比较,(1)P<0.05。

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OPN/IGF1促进小鼠皮质脊髓束轴突再生的作用及机制研究
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张倩 1 , 张坤 2 , 张增强 1 , 李剑勇 1 , 崔芳 1, *
解放军医学杂志 | 基础研究 2021,46(10): 955-960
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解放军医学杂志 | 基础研究 2021, 46(10): 955-960
OPN/IGF1促进小鼠皮质脊髓束轴突再生的作用及机制研究
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张倩1, 张坤2, 张增强1, 李剑勇1, 崔芳1, *
作者信息
  • 1解放军总医院海南医院神经内科,海南三亚 572013
  • 2空军军医大学神经生物学教研室,西安 710032
  • 张倩,博士研究生,主要从事脑血管病、神经系统变性病、周围神经病的诊疗研究

通讯作者:

崔芳,E-mail:
OPN/IGF1 promotes the corticospinal tract regeneration by increasing neuron IGFR expression
Qian Zhang1, Kun Zhang2, Zeng-Qiang Zhang1, Jian-Yong Li1, Fang Cui1, *
Affiliations
  • 1Department of Neurology, Hainan Hospital of Chinese PLA General Hospital, Sanya, Hainan 572013, China
  • 2Department of Neurobiology, Air Force Military Medical University, Xi'an 710032, China
出版时间: 2021-10-28 doi: 10.11855/j.issn.0577-7402.2021.10.01
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目的 研究骨桥蛋白(OPN)/胰岛素样生长因子1(IGF1)在小鼠皮质脊髓束(CST)再生中的作用及相关分子机制。方法 6~8周龄C57BL/6小鼠28只,随机分为实验组与对照组,每组14只。建立小鼠单侧锥体束横断模型,实验组在损伤对侧皮质注射腺相关病毒(AAV)-OPN/IGF1,对照组在损伤对侧皮质注射AAV-胎盘碱性磷酸酶(PLAP)。术后每2周采用行为学方法评估小鼠运动功能的恢复情况。术后12周处死小鼠,采用脊髓蛋白激酶Cγ(PKCγ)免疫组化染色判断皮质脊髓束横断情况;采用红色荧光蛋白(RFP)免疫组化染色分析轴突发芽的情况;采用免疫荧光染色检测神经元OPN和IGF1相关信号通路中核糖体蛋白S6激酶(pS6)和胰岛素样生长因子受体(IGFR)表达情况,研究OPN/IGF1促进轴突发芽的分子机制。结果 通过采用PKCγ免疫组化染色判断皮质脊髓束残余情况,最终实验组入组小鼠10只,对照组入组小鼠8只。实验组与对照组相比,OPN/IGF1对小鼠运动功能恢复无明显作用;皮质神经元高表达OPN/IGF1以后,损伤对侧皮质脊髓束发芽增多,神经元pS6和IGFR表达均明显高于对照组,差异有统计学意义(P<0.05)。结论 OPN/IGF1可通过增加神经元IGFR和pS6表达而促进CST发芽。

骨桥蛋白/胰岛素样生长因子1  /  锥体束横断术  /  轴突再生  /  胰岛素样生长因子受体

Objectives To study the role and associated potential molecular mechanism of osteopontin/insulin-like growth factor 1 (OPN/IGF1) in the corticospinal tract (CST) regeneration. Methods A total of 28 adult C57BL/6 mice (6-8 weeks old) were randomized into the experimental group (injection of AAV-OPN/IGF1, n=14) and control group (injection of AAV-PLAP, n=14) after training of single pellet retrieval. After received the unilateral pyramidotomy surgery, animals received AAV-OPN/IGF1 or AAV-PLAP at the injury site. Further, we employed the movement behaviour test to evaluate spinal motor recovery at 2-week intervals till 12-week after surgery. The protein kinase Cγ (PKCγ) staining in the dorsal funiculus of the cervical spinal cord indicated surgical efficiency. Lastly, we examined the axon sprouting and expression levels of ribosomal protein S6 kinase (pS6)and insulin-like growth factor receptor (IGFR) in neurons using immunohistochemical staining to study the potential mechanism of OPN/IGF1 to promote axon prouting. Results After evaluating PKCγ staining to indicate surgery effectiveness, a total of ten mice were in the experimental group and eight in the control group. In the mouse unilateral pyramidotomy model, OPN/IGF1 has not much effect on the recovery of motor function compared with the control group. After the overexpression of OPN/IGF1 on cortical neurons, sprouting of the spared CST was increased by the elevated expression of pS6 and IGFR, and the difference was statistically significant (P<0.05). Conclusions OPN/IGF1 promotes the sprouting of the CST by increasing the expression of neuronal IGFR and pS6.

osteopontin/insulin-like growth factor 1  /  pyramidotomy  /  axon regeneration  /  insulin-like growth factor receptor
张倩, 张坤, 张增强, 李剑勇, 崔芳. OPN/IGF1促进小鼠皮质脊髓束轴突再生的作用及机制研究. 解放军医学杂志, 2021 , 46 (10) : 955 -960 . DOI: 10.11855/j.issn.0577-7402.2021.10.01
Qian Zhang, Kun Zhang, Zeng-Qiang Zhang, Jian-Yong Li, Fang Cui. OPN/IGF1 promotes the corticospinal tract regeneration by increasing neuron IGFR expression[J]. Medical Journal of Chinese People’s Liberation Army, 2021 , 46 (10) : 955 -960 . DOI: 10.11855/j.issn.0577-7402.2021.10.01
脊髓损伤(spinal cord injury,SCI)及脑卒中后自发性运动功能恢复有限,主要原因是中枢神经系统(central nervous system,CNS)神经元损伤后很难再生[1-2],尤其是皮质脊髓束(corticospinal tract,CST)几乎无再生潜能[3-4]。SCI后有限的功能恢复主要依赖于CST发芽[5],大量研究证实,通过调控相关分子的表达增加神经元的内在再生潜能,从而促进CST再生,有助于损伤后运动功能的恢复[6]
SCI后运动功能恢复通常依赖以下两种方式:损伤后CST再生穿过损伤部位,或未损伤的轴突代偿性发芽,与中间神经元建立联系间接支配失神经的肌肉。鉴于CNS神经元再生能力弱,损伤后的轴突无法重建原始回路,能取得的功能恢复有限[7-8],如何增加CST的发芽以促进功能恢复是目前迫切需要解决的难题。最近,在小鼠脊髓半切和脑卒中模型中的研究显示,信号分子骨桥蛋白(osteopontin,OPN)可增加神经元对胰岛素样生长因子1(insulin-like growth factor 1,IGF1)的敏感性,进而促进CST再生和发芽[9]。本研究采用小鼠单侧锥体束横断模型研究OPN/IGF1促进轴突发芽的分子机制,以期寻找促进CNS损伤后轴突再生的方法。
兔抗小鼠红色荧光蛋白(red fluorescent protein,RFP)多克隆抗体(ab34771,英国Abcam公司);兔抗脊髓蛋白激酶Cγ(protein kinase Cγ,PKCγ)多克隆抗体(sc211)、兔抗小鼠胰岛素样生长因子受体(insulin-like growth factor receptor,IGFR)单克隆抗体(sc-712)(美国Santa Cruz公司);兔抗小鼠核糖体蛋白S6激酶(ribosomal protein S6 kinase,pS6)单克隆抗体(4857,美国Cell Signaling Technology 公司);神经元核抗原(neuronal nuclei antigen,NeuN)小鼠单克隆抗体(MAB377,美国Millipore公司);Alexa FlourR647 标记的山羊抗小鼠IgG(A-21236,美国ThermoFisher Scientific公司);Alexa FlourR405 标记的山羊抗兔IgG(A31556,美国Invitrogen公司);Alexa Fluor 594标记的山羊抗兔IgG(A-11012,美国Life Technology公司)。病毒:AAV-IGF1,AAV-OPN,AAV-胎盘碱性磷酸酶(placental alkaline phosphatase,PLAP),AAV-ChR2-mCherry(红色荧光蛋白,波士顿儿童医院,viral core)。4,6-二脒基-2-苯基吲哚(4,6-diamidino-2-phenylindole,DAPI,28718-90-3,美国Sigma公司),糖丸(F0071,日本Bio-Serv公司);意大利面(pasta);氯胺酮(美国Hospira公司);赛拉嗪(美国AnaSed公司)。冰冻切片机(德国Leica CM3050S);微量注射器(UMC4 Micro4TM Controller);小鼠立体定位仪(美国Stoelting公司);共聚焦显微镜(Zeiss 700和Zeiss 710,日本);高速摄像机(Hotshot e64);不规则水平梯;糖丸抓取箱。
6~8周龄C57BL/6野生型小鼠28只,雌雄不限,体重25~30 g,购自美国Charles River公司。所有实验程序均按照波士顿儿童医院动物使用委员会批准的动物规程(17-05-3451R,Boston Children's Hospital)进行,审批号17-05-3451R。28只小鼠按照随机分配原则均分为实验组与对照组。实验组:单侧锥体束切断,对侧皮质给予AAV-OPN/IGF1和AAV-mCherry注射;对照组:单侧锥体束切断,对侧皮质给予AAV-PLAP和AAV-mCherry注射。结合PKCγ免疫组化染色结果,去除CST残余超过80%的小鼠,最终实验组入组10只,对照组入组8只。各项行为学检测采取录像及双盲方法,实验过程符合国家及单位实验动物管理和使用的相关规定。
氯胺酮(100 mg/kg)联合赛拉嗪(10 mg/kg)麻醉小鼠后,俯卧位固定于离体定位仪上,在相应颅骨上以(±1.5,0)为中心打开颅骨(长4 mm,宽2 mm),实验组给予AAV-IGF1+AAV-OPN+AAV-ChR2-mCherry病毒注射,对照组给予AAV-PLAP+AAV-ChR2-mCherry病毒注射,病毒浓度调至(0.5~5)×1013拷贝/ml。注射坐标位分别为:(±1.3,1.0)(±1.3,0.5)(±1.3,0) (±1.3,-0.5) (±1.3,-1.0)(±1.3,-1.5),每个点注射200 μl。病毒注射皮质是控制小鼠抓取糖丸的上肢对侧皮质(不同小鼠抓取糖丸的优势上肢不同)。病毒注射后1周,再次麻醉小鼠,仰卧位固定,沿气管一侧切开皮肤,钝性分离肌肉,暴露颅底,咬骨钳打开枕骨,暴露延髓,用手术刀在锥体交叉以上横断一侧锥体束(控制小鼠抓取糖丸的优势一侧),逐层缝合肌肉和皮肤。术后每2周进行行为学检测。
术前2周,水平梯训练小鼠行走,用高速摄像机(Hotshot e64)录像,使错误率达到20%左右,表明训练成功。结果判断:小鼠前掌正中央放在小钢柱上,且足趾抓牢为正确,其余均为错误。
小鼠提前禁食18 h,放置于特制透明装置内(203.00 cm×153.00 cm×8.53.00 cm),正前方有一个宽0.53 cm、高133.00 cm 的开口,外加一个可以放置糖丸的平台,20 min内放置40个糖丸,小鼠用优势上肢抓取糖丸计为成功,统计成功率(剔除无抓取糖丸意识或用舌头取糖丸的小鼠)。
将意大利面裁剪成长度为1.5 cm,禁食18 h的小鼠适应环境后,给予一根1.5 cm的意大利面,记录进食完整意大利面的时间。
给予小鼠过量麻醉药后,用4%多聚甲醛固定取材,30%蔗糖溶液脱水24 h后,用组织包埋剂包被组织,冷冻切片(颈段脊髓40 μm,脑组织60 μm),用10%山羊血清+0.5%Triton-100溶液室温下孵育2 h,一抗兔抗PKCγ多克隆抗体(1:100)、兔抗小鼠RFP多克隆抗体(1:400)、兔抗小鼠IGFR单克隆抗体(1:200)、兔抗小鼠pS6单克隆抗体(1:200)、NeuN小鼠单克隆抗体(1:150)4 ℃条件下孵育过夜,对应的二抗(1:1000)室温下孵育2 h,DAPI(1:5000)室温下孵育10 min。共聚焦显微镜在同一条件下采集图像,采用Image J软件选取不同区域NeuN阳性细胞,统计pS6、IGFR荧光强度,取平均值。分析至少统计3套切片。
采用PKCγ免疫组化染色判断皮质脊髓束横断情况;锥体束横断术后12周,将脊髓切片按上述方法进行RFP免疫组化染色。通过脊髓中央导水管及灰质边缘画一水平线,采用3条垂直线Mid、Z1、Z2把轴突损伤侧脊髓灰质的水平线分为3等份,计算通过3条垂直线的轴突数量,得到的数值除以每只小鼠在延髓处(锥体交叉以上)标记的轴突数量(至少选取2套切片,每套计算4个10 000 μm2/个范围内标记到的轴突数量)。
采用GraphPad 5.0和SPSS 16.0软件进行统计分析。计量资料采用$\bar{x}±s$表示,运动行为学分析采用Two-way ANOVA方法,轴突发芽数量和神经元荧光强度的比较采用独立样本t检验。P<0.05为差异有统计学意义。
单侧锥体束横断术后不同时间,采用运动行为学方法评估小鼠运动行为学的恢复情况,结果显示:在不规则水平梯中,术后第2周以后,对照组神经损伤侧的错误率高于实验组,并且持续到术后12周,但两者之间差异无统计学意义(P>0.05);锥体束切断术前及术后,实验组及对照组神经未损伤侧错误率在不同时间点之间均未见明显差异,损伤后相同时间点实验组与对照组比较差异无统计学意义(P>0.05,图1A)。在糖丸抓取行为学测试中,实验组与对照组成功率差异无统计学意义(P>0.05,图1B);在握食意大利面行为学测试中,两组动物完成任务的时间差异无统计学意义(P>0.05,图1C)。
RFP染色结果显示,实验组未损伤侧轴突向损伤侧发芽明显多于对照组(图2A);统计通过Mid、Z1、Z2三条垂直线的轴突数量(图2B),实验组通过Mid、Z1、Z2发芽轴突数量明显多于对照组,差异有统计学意义(P<0.05,图2C)。
对神经元pS6荧光强度进行统计学分析,结果显示,实验组pS6表达明显高于对照组,差异有统计学意义(P<0.05,图3A),表明皮质注射AAV-OPN/IGF1可促进神经元高表达pS6。对神经元IGFR荧光强度进行统计学分析,结果显示,实验组IGFR表达明显高于对照组,差异有统计学意义(P<0.05,图3B),显示皮质注射AAV-OPN/IGF1可促进神经元IGFR的表达。
研究发现,敲除人类第10号染色体缺失的磷酸酶及张力蛋白同源的基因(phosphatase and tension homologue deleted from chromosome10,PTEN),能够增加CNS神经元再生潜能[3,10],PTEN基因敲除后,通过调节雷帕霉素靶蛋白(mechanistic target of rapamycin,mTOR)表达促进轴突再生,但作为抑癌基因PTEN不适用于临床应用。进一步研究发现,PTEN可通过抑制OPN的表达影响mTOR,而OPN存在于各种组织细胞中,把OPN作为调节靶点成为研究热点。在小鼠视神经横断模型中,促进视神经节细胞高表达OPN、IGF1或脑源性神经生长因子(brain derived neurotrophic factor,BDNF)可促进轴突再生,使小鼠恢复部分视力[8,11]。体外培养神经元时给予IGF1或BDNF,可促进其轴突再生[12-14],但在体内实验中,仅给予IGF1或BDNF并不能有效促进CST再生[15],可能是由于成熟神经元对神经生长因子的反应性降低。因此,探寻有效的措施来增加成熟神经元对神经生长因子的敏感性,可能更有利于轴突再生。
SCI后通过中和外源性的抑制分子、增强神经元活性,可以促进CST再生,恢复部分运动功能[7,16]。轴突损伤后,主要通过两种方式再生:被横断的轴突再生越过损伤区;未损伤的轴突发芽,与中间神经元建立联系[17]。因局部环境中的抑制分子以及瘢痕形成,损伤后的轴突很难穿越损伤区;通过去除抑制分子、增强神经元的再生潜能,可以促进未损伤CST代偿性发芽,从而有利于SCI后运动功能的恢复[8-9,18-19]。本课题组前期研究发现,在小鼠T10半切模型和脑卒中模型中,OPN/IGF1可促进轴突的再生和发芽,进而促进运动功能的恢复[9],但在单侧锥体束横断模型中是否具有同样的效果尚不清楚。单侧锥体束横断术主要对前肢的运动功能造成影响,尤其是前肢抓握功能[6,20-21]。因此,本研究选择不规则步行梯、抓取糖丸和握食意大利面测试来评估小鼠运动功能的恢复情况。
在单侧锥体束横断模型中,本研究结果显示OPN/IGF1可促进未损伤CST的发芽(图2),但实验组与对照组小鼠运动功能恢复情况并无明显差异(图1)。推测可能的原因有两种:第一,发芽的轴突尚未与损伤侧神经元建立有效的联系,不能发挥其调节运动功能恢复的作用;第二,因锥体束横断术手术难度大,模型制作过程中可能存在误差,不能完全横断所有锥体束,或切的过深加重损伤,均可能对动物运动行为学的观察造成影响,增加实验误差,导致两组动物行为学无差异。
有研究发现,体外培养原代神经元时,给予IGF1可以促进轴突生长[14]。但体内实验采用小鼠脑卒中模型,单独给予IGF1不能促进损伤后轴突再生[15]。本研究显示,成熟神经元和未成熟神经元对神经营养因子的反应不同,单独应用IGF1或者OPN并不能增加IGFR的表达进而促进轴突再生,而联合应用IGF1和OPN可增加神经元IGFR和pS6的表达[9]。OPN可以提高神经元对IGF1的反应,OPN作为可溶性的蛋白信号分子,作用于细胞表面,通过增加神经元IGFR的表达促进轴突再生(图4)。OPN可通过整合素(integrins)和CD44作用于不同类型的细胞,在皮肤细胞和免疫细胞中,IGFR和整合素均与细胞表面脂蛋白相互连接,向细胞内传递信号[22-23]。基于以上研究结果,推测OPN可能通过整合细胞表面的IGFR调节相关信号通路,进而促进轴突再生。
综上所述,本研究通过神经元高表达OPN/IGF1增加了神经细胞的再生潜能,促进了CST再生,而OPN、IGF1作为可溶性蛋白[9],用于CNS损伤后修复,具有很高的转化医学价值,该研究结果为今后临床应用OPN、IGF1治疗SCI及脑卒中提供了实验依据。
  • 国家自然科学基金(81901252)
  • 国家自然科学基金(81601056)
  • 海南省自然科学基金(820QN383)
  • 海南省自然科学基金(2018CXTD348)
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2021年第46卷第10期
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doi: 10.11855/j.issn.0577-7402.2021.10.01
  • 接收时间:2020-10-25
  • 首发时间:2025-12-19
  • 出版时间:2021-10-28
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  • 收稿日期:2020-10-25
  • 修回日期:2021-06-20
基金
National Natural Science Foundation of China(81901252)
国家自然科学基金(81901252)
National Natural Science Foundation of China(81601056)
国家自然科学基金(81601056)
Natural Science Foundation of Hainan Province(820QN383)
海南省自然科学基金(820QN383)
Natural Science Foundation of Hainan Province(2018CXTD348)
海南省自然科学基金(2018CXTD348)
作者信息
    1解放军总医院海南医院神经内科,海南三亚 572013
    2空军军医大学神经生物学教研室,西安 710032

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2种不同金属材料的力学参数

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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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