摘要
目的探究创伤性脑损伤后表皮生长因子结构域7(EGFL7)及血管生成因子(CD34)在脑组织中的表达及其在血管形成中的作用机制。方法将成年雄性大鼠随机分为研究组和对照组,按损伤后1 h、6 h、24 h、3 d、7 d、14 d分为6个亚组,每组12只。研究组大鼠按创伤性脑损伤进行造模,对照组进行假手术。对两组大鼠进行神经行为学测评;免疫组织化学染色检测大鼠脑组织EGFL7、CD34表达并计算其微血管密度;蛋白质免疫印迹(WB)检测大鼠脑组织EGFL7蛋白表达;实时荧光定量PCR(RT-PCR)检测EGFL7mRNA表达。结果和对照组相比,创伤性脑损伤(TBI)大鼠随受损时间延长,神经行为学评分呈上升趋势(P<0.05),于1~3 d到达高峰(P<0.05);免疫组织化学染色显示,研究组脑组织损伤区有EGFL7、CD34表达,EGFL7阳性细胞数与对照组相比差异有统计学意义(P<0.05),于伤后24 h表达最高,后呈缓慢下降趋势,但高于对照组;研究组的CD34阳性细胞数与对照组相比差异有统计学意义(P<0.05)。于伤后3 d达最高峰,后呈缓慢下降趋势,但仍高于对照组;研究组脑损伤后的大鼠微血管数高于对照组;WB结果示,研究组脑损伤区EGFL7表达的相对灰度值呈升高趋势,与对照组相比差异有统计学意义(P<0.05)。于伤后24 h表达最高;RT-PCR结果示,研究组脑损伤区EGFL7mRNA表达与对照组相比差异有统计学意义(P<0.05)。于伤后3 d时表达最高。结论EGFL7、CD34在创伤性脑组织中表达升高,局部微血管密度增加,主要与新生的血管形成和内皮细胞的正确排列有关,可促进内皮细胞增植、迁移、粘附,促进局部微循环,修复局部受损脑组织。
Objective To explore the expression of EGFL7 and CD34 in brain tissue and the mechanism of angiogenesis after traumatic brain injury(TBI).MethodsAdult male rats were randomly divided into study group and control group,and divided into 6 subgroups according to 1 hr,6 hrs,24 hrs,3 days,7 days,and 14 days after injury,with 12 rats in each group.Rats in the study group were modeled after traumatic brain injury,and the control group underwent sham surgery.Neurobehavioral evaluation was performed on the two groups of rats;the expression of EGFL7 and CD34 in rat brain tissue were measured by immunohistochemical staining and the microvessel density was calculated;EGFL7 protein expression in rat brain tissue was detected by WB.RT-PCR was used to detect EGFL7 mRNA expression.ResultsCompared with the control group,the TBI rats showed an increasing trend in neurobehavioral scores with prolonged injury time(P<0.05),and reached a peak at 1-3 days(P<0.05).EGFL7 and CD34 were expressed in the brain tissue injury area,and the number of EGFL7 positive cells was different from that of the control group(P<0.05).The expression was highest at 24 hrs after injury,and then showed a slowly decreasing trend,but higher than that of the control group.The number of CD34 positive cells was significantly different from that of the control group(P<0.05).After the injury,CD34 reached a peak,and then gradually decreased,but still higher than the control group.The number of microvessels in the study group after brain injury was higher than that in the control group.The degree value was increasing,and there was a difference compared with the control group(P<0.05).The expression was the highest 24 hrs after injury;RT-PCR results showed that the expression of EGFL7 mRNA in the brain injury area of the study group was significantly different from that of the control group(P<0.05).The expression was the highest at 3 days after injury.ConclusionThe expression of EGFL7 and CD34 in traumatic brain tissue is increased,and the local microvessel density is increased,which is mainly related to the formation of new blood vessels and the correct arrangement of endothelial cells,which can promote the proliferation,migration and adhesion of endothelial cells and promote local microcirculation to repair locally damaged brain tissue.
作者
方俊
童文捷
黄波
FANG Jun;TONG Wenjie;HUANG Bo(Department of Neurosurgery, Central Hospital in Songjiang District, Shanghai 201600, China)
出处
《新疆医科大学学报》
CAS
2020年第4期409-413,共5页
Journal of Xinjiang Medical University
基金
上海市卫生健康委科研资助项目(201540204)。
