CAJ | 학술논문

目的探讨多重免疫荧光技术在神经细胞形态学研究中的应用,为神经细胞靶向研究提供新方法. 方法 Balb/c小鼠10只按随机数字表法分为重度创伤性脑损伤(STBI)组、假模对照组(各5只),其中STBI组采用控制性经脑皮质撞击法进行造模,神经功能评级为8分.各组均于第7天终止实验取脑切片,应用多重免疫荧光技术方法,在激光共聚焦显微镜下同时观测神经元核抗原抗体(NeuN)、5-溴脱氧尿嘧啶核苷(BrdU)、胶质纤维酸性蛋白(GFAP)及4',6-二脒基-2-苯基吲哚(DAPI)的形态学表达并定量比较. 结果利用多重免疫荧光技术获得的图像色彩鲜明、清晰明辨、形态感强.STBI组大脑损伤区图像显示BrdU+NeuN+GFAP+DAPI表达较多;假模对照组大脑对应区域图像上几乎没有BrdU+NeuN+DAPI表达,GFAP表达也非常少.STBI组与假模对照组BrdU+NeuN+DAPI和GFAP相对表达量比较差异均有统计学意义(P＜0.05). 结论小鼠遭受重度脑创伤后可诱导神经细胞或星形胶质细胞不同程度自行增殖.多重免疫荧光技术可作为靶向研究神经细胞发育、增殖、凋亡或死亡机制的一种新的科学研究手段.
목적 탐토다중면역형광기술재신경세포형태학연구중적응용,위신경세포파향연구제공신방법. 방법 Balb/c소서10지안수궤수자표법분위중도창상성뇌손상(STBI)조、가모대조조(각5지),기중STBI조채용공제성경뇌피질당격법진행조모,신경공능평급위8분.각조균우제7천종지실험취뇌절편,응용다중면역형광기술방법,재격광공취초현미경하동시관측신경원핵항원항체(NeuN)、5-추탈양뇨밀정핵감(BrdU)、효질섬유산성단백(GFAP)급4',6-이미기-2-분기신타(DAPI)적형태학표체병정량비교. 결과 이용다중면역형광기술획득적도상색채선명、청석명변、형태감강.STBI조대뇌손상구도상현시BrdU+NeuN+GFAP+DAPI표체교다;가모대조조대뇌대응구역도상상궤호몰유BrdU+NeuN+DAPI표체,GFAP표체야비상소.STBI조여가모대조조BrdU+NeuN+DAPI화GFAP상대표체량비교차이균유통계학의의(P＜0.05). 결론 소서조수중도뇌창상후가유도신경세포혹성형효질세포불동정도자행증식.다중면역형광기술가작위파향연구신경세포발육、증식、조망혹사망궤제적일충신적과학연구수단.
Objective To explore the application of multi-overlap immunofluorescence techniques in the morphological research of nerve cells,and provide a new method of targeted research on nerve cells.Methods Ten Balb/c mice were selected and randomly divided into experimental group of severe traumatic brain injury (STBI) and sham control group (n=5);mice in the experimental group were induced STBI models by controlled cortical impact (CCI) method,and the neurological severity scale (NSS) scores were 8 in them.All animals were terminated on day 7.The gene and protein expressions of neuronal nuclei antigen (NeuN),glial fibrillary acidic protein (GFAP) and bromodeoxyuridine (BrdU),and the morphology expression of4',6-diamidino-2-phenylindole (DAPI) were observed under confocal microscopy by applying multi-overlap immunofluorescence techniques.Results The images got by multi-overlap immunofluorescence techniques possessed the characteristics of bright and clear color,strong morphological sense on the expressions.The more obvious expressions of BrdU+NeuN+GFAP+DAPI in the STBI group could be noted as compared with those in the sham control group;the relative expression quantity of BrdU+NeuN+DAPI was significantly different between the two groups (P＜0.05).Conclusion STBI in mice could induce self-regenerated phenomenon on cerebral neural cells or astrocyte;these new techniques provide new scientific means in the targeted study of development,proliferation,apoptosis or death of nerve cells and its mechanism.