摘要
目的探讨血管内皮生长因子（VEGF）基因转移对6-羟多巴胺（6-OHDA）诱导的多巴胺能细胞损伤的保护作用。方法用腺病毒载体携带VEGF165基因（Ad-VEGF165）感染PC12细胞，并设腺病毒携带的β-半乳糖苷酶基因（Ad-LacZ）感染组和磷酸盐缓冲液（PBS）处理组作对照，基因转移成功后，用6-OHDA处理细胞，另设正常对照（不加6—0HDA）。四甲基偶氮唑盐法（MTT）测定细胞活性，免疫荧光细胞化学法检测细胞酪氨酸羟化酶（TH）表达，高效液相色谱检测细胞分泌功能。结果Ad-VEGF165感染组细胞吸光度A570（0．31±0．07），高于Ad-LacZ感染组（0．15±0．07）和PBS处理组（0．13±0．05），但低于正常对照组（0．55±0．10），分别与各组比较，差异均有统计学意义（均为P＜0．01）；免疫荧光染色显示Ad-VEGF组细胞胞浆平均荧光强度（86．75±21．62）高于Ad-LacZ组（51．53±17．49）及PBS组（54．19±15．82），但低于正常对照组（110．39±24．21），分别与各组比较，差异均有统计学意义（均为（P＜0．05）；Ad-VEGF感染组细胞培养液中多巴胺和去甲肾上腺素含量也高于Ad-LacZ感染组和PBS处理组。结论腺病毒介导的VEGF基因转移对6-OHDA诱导的多巴胺能细胞损伤具有保护作用。
Objective To explore the protective effect of vascular endothelial growth factor165 （VEGF165） gene transfer on dopaminergic cells against 6-hydroxydopamine-induced degeneration. Methods Adenovirus vector carrying the gene of VEGF165 （Ad-VEGF165） was used to infect PC12 cells, and groups of adenovirus vector carrying β-galactosidae gene （Ad-LacZ） and phosphate buffered solution （PBS） were set for controls, and then the cells were exposed to 6-hydroxydopamine （6-OHDA） as the cell model of Parkinson disease. Methyhhiazolyl tetrazolium （MTT） and immunofluorescence-cytochemistry were used to assay the viability of PC12 cell and the expressions of the tyrosine hydroxylase （ TH ） respectively. The high performance liquid chromatography （HPLC-ECD） were performed to detect the secretion function of the PC12 cells. Results MTT showed that the cell absorbance A570 of Ad-VEGF165 group （0.31 ±0. 07） was higher than that of Ad-LacZ group （0. 15±0. 07） and PBS group （0.13±0.05）, but lower than that of normal control （0. 55±0.10） （all P〈 0. 01） ; TH immunofluorescence showed that the mean fluorescence intensity in the A＆VEGF group （86.75±21.62） was higher than that of Ad-LacZ group （51.53±17.49） and PBS group （54. 19±15.82）, but lower than that of normal control （110. 39±24.21） （all P〈0. 05）. HPLC-ECD showed that the levels of dopamine and noradrenaline in the culture fluid of Ad-VEGF165 group were higher than those of Ad-LacZ group and PBS group. Conclusions Gene transfer of Ad-VEGF165 has a protective effect on dopaminergic cells against 6-hydroxydopamine-induced degeneration.
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2007年第2期140-144,共5页
Chinese Journal of Geriatrics
关键词
血管内皮生长因子类
腺病毒
细胞保护
基因治疗法
多巴胺
Vascular endothelial growth factors
Adenovirus, human
Cytoprotection
Gene therapy
Dopamine
