人脐带间充质干细胞对大鼠创伤性脑损伤后血脑屏障的保护  

Protective effect of human umbilical cord blood mesenchymal stem cells on the rat’s blood-brain barrier after traumatic brain injury

作　　者：陈江龙 史新宇 程军 叶益超 张震文 李晓红 孙洪涛 Chen Jianglong;Shi Xinyu;Cheng Jun;Ye Yichao;Zhang Zhenwen;Li Xiaohong;Sun Hongtao(Logistics University of People’s Armed Police Force,Tianjin 300162,China;Institute of TBI and Neuroscience,Characteristic Medical Center of Chinese People’s Armed Police Force(PAP),Tianjin 300162,China;Shaoxing People’s Hospital,Shaoxing 312000,Zhejiang Province,China)

机构地区：[1]武警后勤学院,天津市300162 [2]脑创伤与神经疾病研究所,武警特色医学中心,天津市300162 [3]绍兴市人民医院,浙江省绍兴市312000

出　　处：《中国组织工程研究》2020年第25期3947-3952,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81671222),项目参与人:李晓红;天津市救援医学临床医学研究中心课题任务(15ZXLCSY00040-10),项目负责人:孙洪涛。

摘　　要：背景:人脐带间充质干细胞在创伤性脑损伤后血脑屏障修复过程中发挥着至关重要的作用。目的:探讨人脐带间充质干细胞移植对创伤性脑损伤大鼠血脑屏障的保护作用及可能机制。方法:①将60只SD大鼠随机分为假手术组、脑损伤模型组、人脐带间充质干细胞组及抑制剂Sunitinib组,每组15只。除假手术组外,其他3组采用大鼠可控性皮质撞击仪制作创伤性脑损伤模型;②造模后0.5,24,48 h,假手术组、脑损伤模型组大鼠经尾静脉注射1 mL生理盐水,人脐带间充质干细胞组、抑制剂Sunitinib组大鼠经尾静脉注射2×10^9 L^-1人脐带间充质干细胞1 mL,抑制剂Sunitinib组大鼠从造模前1 d开始至处死每天按80 mg/kg的剂量口服PDGFR-β通路抑制剂Sunitinib;③造模后3 d,利用干湿比重法计算各组大鼠脑组织含水量,伊文思蓝法检测血脑屏障的通透性,免疫荧光染色观察GFAP和vWF的表达,Western blot检测血脑屏障相关蛋白及PDGFR-β蛋白的表达。结果与结论:①与假手术组比较,脑损伤模型组的脑含水量、伊文思蓝含量、vWF和GFAP表达明显升高(P<0.05),ZO-1、Oclaudin-5、PDGFR-β蛋白表达明显降低(P<0.05);②与脑损伤模型组比较,人脐带间充质干细胞组的脑含水量、伊文思蓝含量、vWF和GFAP表达明显降低(P<0.05),ZO-1、Oclaudin-5、PDGFR-β蛋白表达明显升高(P<0.05);使用PDGFR-β抑制剂后人脐带间充质干细胞的治疗作用被明显抑制;③结果表明人脐带间充质干细胞可以降低创伤性脑损伤大鼠血脑屏障的通透性,发挥神经保护作用,其作用机制可能是其提高了损伤区PDGFR-β蛋白的表达。BACKGROUND:Human umbilical cord mesenchymal stem cells play a vital role in the repair of the blood-brain barrier after traumatic brain injury.OBJECTIVE:To investigate the protective effect of human umbilical cord blood mesenchymal stem cell transplantation on the blood-brain barrier after traumatic brain injury in rats and its possible mechanism.METHODS:Sixty Sprague-Dawley rats were randomly divided into sham operation group,injury control group(model group),cell transplantation group and Sunitinib group,with 15 rats in each group.Traumatic brain injury model was established by improved hydraulic impact method in all the groups except for the sham operation group.Rats in the sham operation group and model group were injected with 1 mL of normal saline,and those in the cell transplantation group were injected with 1 mL of 2×10^9/L human umbilical cord blood mesenchymal stem cells.The injection was done via the tail vein at 0.5,24,and 48 hours after modeling.In the Sunitinib inhibitor group,the rats were given oral PDGFR-βpathway inhibitor,Sunitinib(80 mg/kg),from 1 day before modeling until being executed.Three days after modeling,the water content in brain tissue was measured by dry-wet specific gravity method,the permeability of the blood-brain barrier was measured by Evans blue method,expression of GFAP and vWF was observed by immunofluorescence staining and the expression of blood-brain barrier related proteins and PDGFR-βpathway proteins was detected by western blot method.RESULTS AND CONCLUSION:Compared with the sham operation group,the brain water content of the model group increased significantly(P<0.05),while that of the cell transplantation group was significantly lower than that of the model group(P<0.05).The Evans blue content in the model group was significantly higher than that in the sham operation group(P<0.05),while the Evans blue content in the cell transplantation group was significantly lower than that in the model group(P<0.05).Compared with the sham operation group,the expression of vWF

关 键 词：人脐带间充质干细胞 创伤性脑损伤 血脑屏障 脑含水量 PDGFR-β通路 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]