CAJ | 학술논문

目的观察3日龄新生大鼠缺血性脑损伤后脑室下区(subventricular zone,SVZ)新生细胞的变化,探讨未成熟脑缺血性损伤后的内源性修复机制. 方法 3日龄新生SD大鼠32只随机分为实验组和对照组,每组16只.实验组结扎双侧颈总动脉,对照组不结扎.两组大鼠均于术后5～7 d给予5-溴脱氧尿嘧啶(5-bromodeoxyuridine,BrdU),每次50 mg/kg腹腔注射,每12 h 1次×6次.两组大鼠分别于术后14 d、28 d处死取脑,采用免疫荧光染色方法检测BrdU、Ⅲ型β-微管蛋白(class Ⅲ β-tubulin,TuJ1)、少突胶质细胞O抗原-4(oligodendroeyte O antigen-4,O4)以及胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达变化,观察SVZ各类新生细胞的变化. 结果实验组术后14 d和28 d新生神经元(BrdU+/TuJ1+双标阳性细胞)分别为(7800±800)个/视野和(10 700±1400)个/视野、新生少突胶质细胞(BrdU+/O4+双标阳性细胞)分别为(6100±1000)个/视野和(7300±1400)个/视野,新生星形胶质细胞(Brdu+/GFAP+双标阳性细胞)分别为(4500±700)个/视野和(6700±1100)个/视野,均随着时点的延长而增加(P<0.01);且各新生神经元数目较同时点对照组明显增多,差异有统计学意义(P均<0.01). 结论新生大鼠缺血性脑损伤后,SVZ新生神经元、新生少突胶质细胞、新生星形胶质细胞增多,提示未成熟脑SVZ在缺血性损伤后有修复作用.
목적 관찰3일령신생대서결혈성뇌손상후뇌실하구(subventricular zone,SVZ)신생세포적변화,탐토미성숙뇌결혈성손상후적내원성수복궤제. 방법 3일령신생SD대서32지수궤분위실험조화대조조,매조16지.실험조결찰쌍측경총동맥,대조조불결찰.량조대서균우술후5～7 d급여5-추탈양뇨밀정(5-bromodeoxyuridine,BrdU),매차50 mg/kg복강주사,매12 h 1차×6차.량조대서분별우술후14 d、28 d처사취뇌,채용면역형광염색방법검측BrdU、Ⅲ형β-미관단백(class Ⅲ β-tubulin,TuJ1)、소돌효질세포O항원-4(oligodendroeyte O antigen-4,O4)이급효질섬유산성단백(glial fibrillary acidic protein,GFAP)적표체변화,관찰SVZ각류신생세포적변화. 결과 실험조술후14 d화28 d신생신경원(BrdU+/TuJ1+쌍표양성세포)분별위(7800±800)개/시야화(10 700±1400)개/시야、신생소돌효질세포(BrdU+/O4+쌍표양성세포)분별위(6100±1000)개/시야화(7300±1400)개/시야,신생성형효질세포(Brdu+/GFAP+쌍표양성세포)분별위(4500±700)개/시야화(6700±1100)개/시야,균수착시점적연장이증가(P<0.01);차각신생신경원수목교동시점대조조명현증다,차이유통계학의의(P균<0.01). 결론 신생대서결혈성뇌손상후,SVZ신생신경원、신생소돌효질세포、신생성형효질세포증다,제시미성숙뇌SVZ재결혈성손상후유수복작용.
Objective To investigate the differentiation of neural stem cells in the subventrieular zone(SVZ)of the 3-day-old neonatal rats with ischemic brain injury and to provide the experimental evidences on endogenous repair mechanism of premature brain injury after ischemia. Methods Thirty-two 3-day-old rats were divided into experimental group and the control group.Rats in the experimental group were subjected to bilateral common arteries occlusion.while those in control group were not.All rats were administrated 5-bromodeoxyuridine(BrdU)50 mg/kg by intraperitoneal injection twice daily during 5 to 7 days after the operation.At 14 days and 28 days after operation,rats were sacrificed and their brains were collected.The changes of newborn cells in the SVZ of brain were observed by marking the BrdU,neuronal class Ⅲ β-tubulin(TuJ1),oligodendrocyte O antigen-4 (O4)and glial fibrillary acidic protein(GFAP)with immunofluorescence. Results Compared with the control group,the number(7800±800 vs 4200±700,10 700±1400 vs 4600±600)of newborn neuron(BrdU+/TuJl+)and the number(6100±1000 vs 2600±500,7300±1400 vs 2800±800)of newborn oligodendrocytes(BrdU+/O4+)per field increased significantly at 14 days and 28 days after operation in the experimental group,so did the number of newborn astrocytes per field(4500±700 vs 2700±500,6700±1100 vs 3000±600)respectively(P<0.01). Conclusions The number of neurons,oligodendroeytes and atrocytes in the SVZ increased after brain ischemia,which suggested that SVZ might have the function of repair after brain ischemia injury.