神经解剖学杂志
神經解剖學雜誌
신경해부학잡지
CHINESE JOURNAL OF NEUROANATOMY
2005年
6期
569-575
,共7页
胰岛素样生长因子1%β样淀粉蛋白%细胞凋亡%Tau%流式细胞%Alzheimer病
胰島素樣生長因子1%β樣澱粉蛋白%細胞凋亡%Tau%流式細胞%Alzheimer病
이도소양생장인자1%β양정분단백%세포조망%Tau%류식세포%Alzheimer병
insulin-like growth factor 1%β-amyloid%apoptosis%tau%flow cytometry%Alzheimer's disease
本研究的目的是阐明胰岛素样生长因子1(IGF-1)对β样淀粉蛋白(Aβ)引起的神经元凋亡的保护作用,以及tau蛋白磷酸化的作用.用MTT(四甲基偶氮唑盐)方法检测细胞活性,用流式细胞学结合Annexin V-FITC和PI(碘化丙锭)双染的方法检测早期凋亡和晚期凋亡/坏死,用Hoechst 33342染色观察凋亡细胞形态学,用免疫细胞化学的方法检测tau蛋白磷酸化.IGF-1阻止了Aβ25-35引起的培养的大鼠海马神经元的毒性,MTT值显著增加,从54.51%增至61.8%,Hoechst 33342阳性细胞的百分比从30.77%减少到22.81%.Aβ25-35孵育使Annexin V单标记细胞(Annexin V+/PI-)以及Annexin V/PI双标记细胞(Annexin V+/PI+)的百分比显著增加(分别为3.41%和19.47%),应用100 ng/ml的IGF-1可显著减少Annexin V单标记细胞和Annexin V/PI双标记细胞的百分比分别至2.98%和15.16%.Aβ25-35可增加tau蛋白磷酸化,AT8阳性细胞占41.84%,而IGF-1则可抑制这一效应.我们的结果表明IGF-1可保护神经元,降低Aβ的细胞毒性,减少早期和晚期凋亡/坏死细胞的比例,抑制tau蛋白磷酸化,这可能是IGF-1神经保护作用的细胞机制.
本研究的目的是闡明胰島素樣生長因子1(IGF-1)對β樣澱粉蛋白(Aβ)引起的神經元凋亡的保護作用,以及tau蛋白燐痠化的作用.用MTT(四甲基偶氮唑鹽)方法檢測細胞活性,用流式細胞學結閤Annexin V-FITC和PI(碘化丙錠)雙染的方法檢測早期凋亡和晚期凋亡/壞死,用Hoechst 33342染色觀察凋亡細胞形態學,用免疫細胞化學的方法檢測tau蛋白燐痠化.IGF-1阻止瞭Aβ25-35引起的培養的大鼠海馬神經元的毒性,MTT值顯著增加,從54.51%增至61.8%,Hoechst 33342暘性細胞的百分比從30.77%減少到22.81%.Aβ25-35孵育使Annexin V單標記細胞(Annexin V+/PI-)以及Annexin V/PI雙標記細胞(Annexin V+/PI+)的百分比顯著增加(分彆為3.41%和19.47%),應用100 ng/ml的IGF-1可顯著減少Annexin V單標記細胞和Annexin V/PI雙標記細胞的百分比分彆至2.98%和15.16%.Aβ25-35可增加tau蛋白燐痠化,AT8暘性細胞佔41.84%,而IGF-1則可抑製這一效應.我們的結果錶明IGF-1可保護神經元,降低Aβ的細胞毒性,減少早期和晚期凋亡/壞死細胞的比例,抑製tau蛋白燐痠化,這可能是IGF-1神經保護作用的細胞機製.
본연구적목적시천명이도소양생장인자1(IGF-1)대β양정분단백(Aβ)인기적신경원조망적보호작용,이급tau단백린산화적작용.용MTT(사갑기우담서염)방법검측세포활성,용류식세포학결합Annexin V-FITC화PI(전화병정)쌍염적방법검측조기조망화만기조망/배사,용Hoechst 33342염색관찰조망세포형태학,용면역세포화학적방법검측tau단백린산화.IGF-1조지료Aβ25-35인기적배양적대서해마신경원적독성,MTT치현저증가,종54.51%증지61.8%,Hoechst 33342양성세포적백분비종30.77%감소도22.81%.Aβ25-35부육사Annexin V단표기세포(Annexin V+/PI-)이급Annexin V/PI쌍표기세포(Annexin V+/PI+)적백분비현저증가(분별위3.41%화19.47%),응용100 ng/ml적IGF-1가현저감소Annexin V단표기세포화Annexin V/PI쌍표기세포적백분비분별지2.98%화15.16%.Aβ25-35가증가tau단백린산화,AT8양성세포점41.84%,이IGF-1칙가억제저일효응.아문적결과표명IGF-1가보호신경원,강저Aβ적세포독성,감소조기화만기조망/배사세포적비례,억제tau단백린산화,저가능시IGF-1신경보호작용적세포궤제.
The aim of this study is to elucidate the protective and anti-apoptotic effects of insulin-like growth factor 1 ( IGF-1 ) against β-amyloid (Aβ) and investigate the effect of IGF-1 on Aβ-induced tau phosphorylation. Cell viability was measured using the MTT (3-(4,5-dimethylthiazolyl-2 )-2,5-diphenyltetrazolium bromide) assay, early apoptosis and late apoptosis/necrosis were analyzed by flow cytometry using Annexin V-FITC and propidium iodide (PI) double staining, and morphology was examined by Hoechst 33342 staining. Tau phosphorylation was detected using AT8 immunostaining. Preincubation of cultured rat hippocampal neurons with IGF-1 for 24 h prevented cytotoxicity induced by Aβ25-35 for 48 h. The MTT value significantly increased from 54.51% to 61.8% of the control group, and the percentage of Hoechst 33342-positive cells decreased from 30.77% to 22.81%. Incubation with Aβ25-35 for 48 h caused a marked increase in the percentages of Annexin V-FITC single-labeled cells (Annexin V +/PI-) and Annexin V/PI double-stained cells (Annexin V +/PI + ) (3.41% and 19.47% , respectively), which were significantly decreased by pretreatment with 100 ng/ml of IGF-1 for 24 h (to 2.98% and 15.16% , respectively). Aβ25-35 treatment increased tau phosphorylation and AT8 positive cells were 41.84%. This effect could be inhibited by different concentrations of IGF-1. Our findings showed that IGF-1 protected against Aβ-induced cytotoxicity, decreased the percentage of early and late apoptosis/necrosis cells, and inhibited tau phosphorylation, which may be the cellular mechanisms for its neuroprotective action.
