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[1]李盟,赵剑,万志远,等.黄芪甲苷对TNF-α上调星形胶质细胞AQP-4基因表达的影响[J].宁夏医科大学学报,2016,(03):240-244.[doi:10.16050/j.cnki.issn1674-6309.2016.03.003]
 LI Meng,ZHAO Jian,WAN Zhiyuan,et al.Influence of Astragaloside IV Intervention on the AQP-4 Gene Expression Induced by TNF-α in Cultured Astrocytes[J].Ningxia Medical University,2016,(03):240-244.[doi:10.16050/j.cnki.issn1674-6309.2016.03.003]
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黄芪甲苷对TNF-α上调星形胶质细胞AQP-4基因表达的影响(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2016年03期
页码:
240-244
栏目:
论著
出版日期:
2016-03-30

文章信息/Info

Title:
Influence of Astragaloside IV Intervention on the AQP-4 Gene Expression Induced by TNF-α in Cultured Astrocytes
作者:
李盟1赵剑1万志远2曹羿堃1王效军1马全瑞1秦毅1
1.宁夏医科大学基础医学院人体解剖与组织胚胎学系,银川 750004; 2.解放军第513医院,酒泉 735000
Author(s):
LI Meng1 ZHAO Jian1 WAN Zhiyuan2 CAO Yikun1 WANG Xiaojun1 MA Quanrui1 QIN Yi1
1.Dept. of Human Anatomy and Histoembryology, School of Basic Medicine Science, Ningxia Medical University, Yinchuan 750004; 2.The PLA`s 513 Hospital, Jiuquan 735000
关键词:
黄芪甲苷 AQP-4 TNF-α 星形胶质细胞
Keywords:
AQP-4 TNF-α astragaloside IV astrocytes
分类号:
R743.33
DOI:
10.16050/j.cnki.issn1674-6309.2016.03.003
文献标志码:
A
摘要:
目的 探讨黄芪甲苷对TNF-α上调星形胶质细胞上AQP-4基因表达的影响。方法 选用第2代第1天的星形胶质细胞。将星形胶质细胞接种到96孔板,分别用0、0.01、0.1、1、10和100ng·mL-1的TNF-α和0、0.001、0.01和0.1μmol·mL-1黄芪甲苷进行干预。用CCK-8试剂盒检测星形胶质细胞活性的变化,以筛选干预用TNF-α和黄芪甲苷的浓度。将星形胶质细胞分别用0、0.5、5、10、30和50ng·mL-1的TNF-α进行干预,用RT-PCR方法检测AQP-4基因表达水平的变化,观察TNF-α对AQP-4基因表达水平的影响。将星形胶质细胞分为Sham组、TNF-α组、0.001μmol·mL-1ASIV+TNF-α组、0.01μmol·mL-1ASIV+TNF-α组、0.1μmol·mL-1ASIV+TNF-α组,用RT-PCR方法检测AQP-4基因表达水平的变化,观察黄芪甲苷对TNF-α诱导星形胶质细胞AQP-4基因表达的影响。结果 当TNF-α的浓度为0.01、0.1、1、10ng·mL-1时,星形胶质细胞的活性与0ng·mL-1相比差异无统计学意义(P<0.05); 当TNF-α的浓度为100ng·mL-1时,星形胶质细胞的活性与0ng·mL-1相比明显下降(P<0.05)。当黄芪甲苷的浓度为0.001、0.01、0.1μmol·mL-1时,星形胶质细胞的活性与0μmol·mL-1相比均有提高(P<0.05)。与0ng·mL-1组相比,各TNF-α干预组的AQP-4基因表达水平均升高(P<0.05),以10ng·mL-1 TNF-α干预组的作用最明显。0.001、0.01、0.1μmol·mL-1黄芪甲苷+TNF-α三个干预组的AQP-4基因表达水平与TNF-α干预组相比明显降低(P<0.05)。结论 当TNF-α的浓度在0~100ng·mL-1之间时对星形胶质细胞无明显细胞毒性; 所选浓度范围的黄芪甲苷可促进星形胶质细胞细胞增殖; TNF-α诱导AQP-4的基因表达具有浓度依赖性; 黄芪甲苷可下调由TNF-α所致的星形胶质细胞上AQP-4基因表达水平的升高。
Abstract:
Objective To examine the effects of astragaloside IV on AQP-4 gene expression induced by TNF-α in cultured astrocytes. Methods 100μL of cell suspension(20000 cells /well)were dispensed in a 96-well plate. Cells were incubated with various concentrations of TNF-α(0,0.01,0.1,1,10,100ng·mL-1)and various concentrations of astragaloside IV(0.001,0.01,0.1μmol·mL-1)separately, and cell activity was measured by cell counting kit-8(CCK-8)to filtrate the concentrations of astragaloside IV and TNF-α. Cell were incubated with various concentrations of TNF-α(0,0.5,5,10,30,50ng·mL-1, and AQP-4 gene level was assessed by qRT-PCR to explore the effects of TNF-α on AQP-4 gene expression. Cells were divided into five groups: sham group, TNF-α group, 0.001 μmol·mL-1 ASIV+TNF-α group, 0.01μmol·mL-1 ASIV+TNF-α group, 0.1μmol·mL-1ASIV +TNF-α group, and AQP-4 gene level was assessed by qRT-PCR to explore the roles of astragaloside IV. Results Compared with the 0ng·mL-1 group, the activity of cells was higher and had a tendency to rise, without statistical significance, when the concentration of TNF-α ranged from 0 to 10 ng·mL-1. When the concentration of TNF-α reached to 100 ng·mL-1, the activity of cells declined significantly(P<0.05)compared with the 0ng·mL-1 group. When the concentration of astragaloside IV was 0.001,0.01,0.1μmol·mL-1, the activity of astrocytes was significantly improved at a dose-dependent manner compared with the 0μmol·mL-1group(P<0.05). qRT-PCR analysis showed that compared with the 0ng·mL-1 group, the expression levels of AQP-4mRNA in astrocytes treated with various concentrations of TNF-α were higher significantly and when the concentration of TNF-α was 10ng·mL-1. The expression levels of AQP-4mRNA in astrocytes treated with 0.001,0.01,0.1μmol·mL-1 ASIV+ TNF-α were significantly lower than that of the TNF-α group. Conclusion TNF-α elevates AQP-4 gene expression in a concentration-dependent manner. Astragaloside IV could inhibit AQP-4 gene expression induced by TNF-α.

参考文献/References:

[1] 沈慧, 肖培, 李信晓, 等. 脑水肿治疗新见解[J].新乡医学院学报, 2014(6):488-491,495.
[2] Xi G, Keep RF, Hoff JT. Mechanisms of brain injury after intracerebral haemorrhage[J].The Lancet Neurology, 2006, 5(1):53-63.
[3] Sun MC, Honey CR, Berk C, et al. Regulation of aquaporin-4 in a traumatic brain injury model in rats[J].J Neurosurg, 2003, 98(3):565-569.
[4] Fukuda AM, Badaut J. Aquaporin 4:a player in cerebral edema and neuroinflammation[J].Journal of Neuroinflammation, 2012, 9(1):279.
[5] Yang B, Zador Z, Verkman AS. Glial cell aquaporin-4 overexpression in transgenic mice accelerates cytotoxic brain swelling[J].J Biol Chem, 2008, 283(22):15280-15286.
[6] Li C, Zhang X, Li H, et al. Brain edema after intracerebral hemorrhage in rats:The role of inflammation[J].Neurol India, 2006, 54(4):402.
[7] Laird MD, Sukumari-Ramesh S, Swift AE, et al. Curcumin attenuates cerebral edema following traumatic brain injury in mice:a possible role for aquaporin-4[J].J Neurochem, 2010, 113(3):637-648.
[8] Alexander JJ, Jacob A, Cunningham P, et al. TNF is a key mediator of septic encephalopathy acting through its receptor, TNF receptor-1[J].Neurochemistry International, 2008, 52(3):447-456.
[9] 赵剑, 刘广辉, 梁亮, 等. 黄芪甲苷对大鼠脑出血后脑水肿及水通道蛋白4表达的影响[J].宁夏医科大学学报, 2015, 37(8):879-882,886.
[10] Inaji M, Tomita H, Tone O, et al. Chronological changes of perihematomal edema of human intracerebral hematoma[J].Acta Neurochir Suppl, 2003, 86:445-448.
[11] Gebel JM Jr, Jauch EC, Brott TG, et al. Relative edema volume is a predictor of outcome in patients with hyperacute spontaneous intracerebral hemorrhage[J].Stroke, 2002, 33(11):2636-2641.
[12] Tait M, Saadoun S, Bell B, et al. Water movements in the brain:role of aquaporins[J].Trends in Neurosciences, 2008(1):37-43.
[13] Kimelberg HK. Current concepts of brain edema. Review of laboratory investigations[J].J Neurosurg, 1995, 83(6):1051-1059.
[14] MacAulay N, Zeuthen T. Water transport between CNS compartments:contributions of aquaporins and cotransporters[J].Neuroscience, 2010, 168(4):941-956.
[15] Nielsen S, Nagelhus EA, Amiry-Moghaddam M, et al. Specialized membrane domains for water transport in glial cells:high-resolution immunogold cytochemistry of aquaporin-4 in rat brain[J].J Neurosci, 1997, 17(1):171-180.
[16] Jung JS, Bhat RV, Preston GM, et al. Molecular characterization of an aquaporin cDNA from brain:candidate osmoreceptor and regulator of water balance[J].Proc Natl Acad Sci USA, 1994, 91(26):13052-13056.
[17] Griesdale DE, Honey CR. Aquaporins and brain edema[J].Surg Neurol, 2004, 61(5):418-421.
[18] Papadopoulos MC, Verkman AS. Aquaporin-4 and brain edema[J].Pediatr Nephrol, 2007, 22(6):778-784.
[19] Manley GT, Binder DK, Papadopoulos MC, et al. New insights into water transport and edema in the central nervous system from phenotype analysis of aquaporin-4 null mice[J].Neuroscience, 2004, 129(4):983-991.
[20] 房金勇. 高压氧对实验性脑出血大鼠出血灶周围水肿及AQP-4表达的影响[D]. 河北医科大学, 2011.
[21] Ding Z, Zhang J, Xu J, et al. Propofol administration modulates AQP-4 expression and brain edema after traumatic brain injury[J].Cell Biochem Biophys, 2013, 67(2):615-622.

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备注/Memo

备注/Memo:
收稿日期:2015-11-16 基金项目:宁夏自然科学基金(NZ14064) 作者简介:李盟,女,在读硕士研究生,研究方向:脑出血病理机制。 通信作者:秦毅,教授,硕士研究生导师。E-mail:qinyi966@163.com
更新日期/Last Update: 2016-03-20