颅骨及硬脑膜中FAK/Twist1在SD大鼠颅脑生长发育过程中作用的研究  

Study on the role of FAK/Twist1 in skull and dura mater during brain growth and development in SD rats

作　　者：王雷 张晓璐 文平 张司玺 杨军 赵志军[1,4] 张春阳 Wang Lei;Zhang Xiaolu;Wen Ping;Zhang Sixi;Yang Jun;Zhao Zhijun;Zhang Chunyang(Department of Neurosurgery,The First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science&Technology,Baotou 014010,China;No.903 Hospital of the Joint Service Support Force,Hangzhou 310000,China;Department of Neurosurgery,Peking University Third Hospital,Beijing 100191,China;Institute of Neurosurgical Diseases,Baotou Medical College(Translational Medicine),Baotou 014040,China;Inner Mongolia Autonomous Region Bone Tissue Regeneration and Injury Repair Engineering Technology Center,Baotou 014040,China)

机构地区：[1]内蒙古科技大学包头医学院第一附属医院神经外科,包头014010 [2]联勤保障部队第九○三医院,杭州310000 [3]北京大学第三医院神经外科,北京100191 [4]包头医学院神经外科疾病研究所(转化医学),包头014040 [5]内蒙古自治区骨组织再生与损伤修复工程技术中心,包头014040

出　　处：《中华神经外科杂志》2024年第7期725-732,共8页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金(82160250,81960238);内蒙古自然科学基金(2021MS08124);国家临床重点专科建设项目(2022-内05);包头医学院研究生教育创新计划项目(bycx2021014)。

摘　　要：目的探究快速生长发育期SD大鼠颅骨的生长发育及其与颅骨和硬脑膜中局部黏着斑激酶(FAK)/Twist1信号通路的关系。方法选取幼龄大鼠,从出生后1周开始,使用软尺记录颅骨长度(颅长)及宽度(颅宽);分别使用电子秤和电子天平称量体质量和脑质量;使用量筒称量脑组织的容积;蛋白质免疫印迹法(WB)检测颅骨FAK和硬脑膜Twist1的表达变化;免疫组织化学(简称免疫组化)和免疫荧光染色法观察颅骨与硬脑膜组织的细胞形态及FAK/Twist1在不同部位的表达差异。结果(1)大鼠颅长、颅宽在1~7周内均逐渐增加(F颅长=420.20、F颅宽=47.27,均P<0.001),与第1周相比,第2周时颅长和颅宽均明显增加(均P<0.05),第7周时颅长和颅宽增长分别占整个发育过程(1~12周)的94.4%和90.0%。(2)大鼠的体质量和脑质量在1~7周内逐渐增加(F体质量=485.20、F脑质量=126.00,均P<0.001),与第1周相比,第2周时体质量和脑质量均明显增加(均P<0.05),第7周时增加的体质量和脑质量分别占整个发育过程的60.4%和51.2%。(3)大鼠的脑容积在1~7周内逐渐增加(F=852.80,P<0.001),1~3周增长迅速,3周以后较为平缓,第7周时增加的脑容积占整个发育过程的60.5%。(4)WB结果显示,第1~7周时颅骨组织中FAK表达水平逐渐升高(F=15.80,P<0.001),第7周时FAK表达水平较第1周明显升高(P<0.01);硬脑膜组织中FAK表达水平呈现先降低后升高的趋势,第4周FAK表达水平达到最高值,与第1周相比,差异有统计学意义(P<0.05);颅骨组织中FAK的表达水平与颅长、颅宽的相关系数分别为0.95、0.96(均P<0.001),呈正相关。(5)颅骨颅缝观察结果显示,出生后第1、4、7周,大鼠的颅骨生长以纵向生长为主,且颅缝逐渐清晰,在第7周时已完全闭合。(6)免疫组化及免疫荧光结果显示,Twist1主要在硬脑膜中表达,在第1周时表达水平最高,随后逐渐下降(F组化=22.10、F荧光=36.82,均P<0.01);FAK主要在颅骨中表达,第4�Objective To investigate the growth and development of skull in rapidly growing SD rats and it′s relationship to FAK-Twist1 signaling pathway in the skull and dura mater.Methods We selected young rats and started from 1 week after birth to document the cranial length and cranial width using a soft ruler.An electronic scale and electronic balance were used to weigh the body weight and brain weight of rats.A graduated cylinder was used to weigh the volume of brain tissue.Western blotting was performed to detect the expression changes of FAK in the skull and Twist1 in the dura mater.Immunohistochemistry and immunofluorescence were used to observe the cell morphology of the skull and dura mater tissues and the expression differences of FAK-Twist1 in different parts.Results(1)The cranial length and width of the rat skull increased gradually from 1 to 7 weeks(Fcranial length=420.20、Fcranial width=47.27,both P<0.001).Compared with the first week,the length and width of the skull increased significantly in the second week(all P<0.05),and the two lines increased by 94.4%and 90.0%of the whole skull development(1-12 weeks)respectively in the seventh week;(2)The body weight and brain weight of rats increased gradually from 1 to 7 weeks(Fbody weight=485.20、Fbrain weight=126.00,both P<0.001).Compared with the first week,there was a significant increase in body weight and brain weight in the second week(all P<0.05).In the seventh week,body weight and brain weight increased by 60.4%and 51.2%of the whole development respectively;(3)The brain volume of rats increased gradually from 1 to 7 weeks(F=852.80,P<0.001),rapidly from 1 to 3 weeks,and relatively slowly after 3 weeks.Compared with the first week,the brain volume increased by 60.45%of the whole development in the seventh week;(4)The results of WB showed,at weeks 1-7,the expression of FAK in the skull tissue gradually increased(F=15.80,P<0.001),and FAK was significantly increased in week 7 compared with week 1(P<0.01).FAK in the dura mater tissue showed a trend of first

关 键 词：颅骨 生长 硬脑膜 局部黏着斑激酶 Twist 1 大鼠 

分 类 号：R-332[医药卫生]