[1]唐兆华,廖正步,谢延风,等.氧糖剥夺、复氧后损伤星形胶质细胞中MMP-9的表达变化与意义[J].陆军军医大学学报(原第三军医大学学报),2012,34(05):387-390.
 Tang Zhaohua,Liao Zhengbu,Xie Yanfeng,et al.Significance of MMP-9 expression in astrocytes after oxygen-glucose deprivation/reoxygenation in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(05):387-390.
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氧糖剥夺、复氧后损伤星形胶质细胞中MMP-9的表达变化与意义(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第05期
页码:
387-390
栏目:
论著
出版日期:
2012-03-15

文章信息/Info

Title:
Significance of MMP-9 expression in astrocytes after oxygen-glucose deprivation/reoxygenation in vitro
作者:
唐兆华廖正步谢延风石全红何朝晖詹彦
重庆医科大学附属第一医院神经外科
Author(s):
Tang Zhaohua Liao Zhengbu Xie Yanfeng Shi Quanhong He Zhaohui Zhan Yan
Department of Neurosurgery, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
基质金属蛋白酶-9星形胶质细胞氧糖剥夺、复氧
Keywords:
matrix metalloproteinase-9 astrocyte oxygen-glucose deprivation/reoxygenation
分类号:
R329.6;R338.1;R345
文献标志码:
A
摘要:
目的      通过观察金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在氧糖剥夺、复氧培养后损伤星形胶质细胞中基质的表达变化并抑制其活性,探讨其在氧糖剥夺、复氧后细胞损伤中的意义。      方法        将星形胶质细胞分为正常组、模型组和GM6001组。模型组细胞在无糖DMEM培养基,1%O2的培养条件下行3、5、7 h时长的氧糖剥夺后复氧至24 h。GM6001组细胞在5 h氧糖剥夺、复氧的过程中加入外源性基质金属蛋白酶抑制剂(GM6001,10 μmol/ml),复氧至24 h。用RT-PCR测定不同时长氧糖剥夺后星形胶质细胞MMP-9 mRNA表达变化,ELISA法测定细胞培养上清液中MMP-9蛋白的浓度变化;用光学显微镜观察各组的细胞形态变化,LDH漏出率、CCK-8法检测细胞的损伤情况及存活率。      结果        ①RT-PCR及ELISA法检测示:与正常组相比,氧糖剥夺、复氧后,模型组细胞MMP-9 mRNA表达量和细胞培养上清液中MMP-9蛋白的浓度均有所增加(P<0.05),随着氧糖剥夺时间的延长,均呈现不断上升趋势;②与正常组相比,氧糖剥夺、复氧后,模型组细胞明显损伤,随着氧糖剥夺时间的延长,细胞损伤加重,光镜下细胞出现明显肿胀,并有部分细胞呈凝固性坏死,LDH漏出率不断增高(P<0.01)、存活率则逐渐下降(P<0.05, P<0.01),且培养上清MMP-9蛋白浓度与细胞LDH漏出率的增高程度呈正相关(r=0.693,P<0.05);与模型组相比,5 h氧糖剥夺、复氧后,GM6001组细胞形态学损伤表现明显减轻,LDH漏出率水平也有明显下降(P<0.01),细胞存活率则显著上升(P<0.05)。      结论        MMP-9过表达可能是氧糖剥夺、复氧后引起细胞损伤的重要因素之一。
Abstract:
Objective        To investigate the significance of matrix metalloproteinase-9 (MMP-9) expression in astrocytes after oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro.       Methods        Astrocytes were divided into control group, OGD/R group and GM6001 group. Astrocytes in OGD/R group were treated with OGD for 3, 5, and 7 h, respectively, and then reoxygenated for 24 h. In GM6001 group, astroctyes were treated with the inhibitor of matrix metalloproteinase (GM6001, 10 μmol/ml) during 5 hours’ OGD/R. The expression of MMP-9 at mRNA and protein levels in the astrocytes were detected after 24 hours’ reoxygenation by RT-PCR and ELISA. The morphological alterations of cells in each group were observed by light microscopy. The cell injury was inspected by LDH activity (LDH %) and Cell Counting Kit-8 (CCK-8).       Results      Compared with normal group, as the OGD time prolonged, the expression of MMP-9 at mRNA and protein levels were increased gradually (P<0.05) after OGD/R, which were further increase with time elapse. OGD resulted in in significant swelling astrocytes and obviously injured with time prolonged, and some cells seemed to be coagulation necrosis, the LDH % was increased (P<0.01) and the survival rate was decreased severely (P<0.05, P<0.01). Protein level of MMP-9 in the supernatant was positively correlated with LDH % (r=0.693, P<0.05). Compared with OGD/R group, cell injury following 5 hours’ OGD/R was remarkably decreased, as survival rate was raised significantly.       Conclusion        Over-expression of MMP-9 might be a important factor in OGD/R-induced cell injury.

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更新日期/Last Update: 2012-03-01