多巴胺对C8胶质细胞生长及LMO3和CIB1表达的影响
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摘要
目的了解多巴胺对C8胶质细胞生长的影响、可能受体途径以及对CIB1、LMO3表达的影响。方法采用MTT方法检测多巴胺、多巴胺受体激动剂、多巴胺受体拮抗剂对C8胶质细胞生长增殖的作用,同时采用免疫荧光组化法检测不同浓度多巴胺对C8胶质细胞中LMO3、CIB1表达及定位的影响。结果多巴胺对C8胶质细胞的作用与浓度相关,低浓度(10μmol/L)对C8胶质细胞生长无明显影响,中浓度(100μmol/L)可明显促进C8胶质细胞的增殖(P<0.05,P<0.01),而高浓度(500μmol/L)则抑制C8胶质细胞生长(P<0.05);多巴胺处理组比多巴胺D1激动剂组促C8胶质细胞生长作用明显,同时加入多巴胺及D2受体拮抗剂组比多巴胺处理组C8胶质细胞显著增加(P<0.01)。低浓度(10μmol/L)多巴胺显著增加C8胶质细胞LMO3的表达水平(P<0.05);高浓度(500μmol/L)多巴胺则降低CIB1蛋白的表达(P<0.05)。结论多巴胺主要通过多巴胺D2受体对C8胶质细胞生长增殖起作用,并呈浓度相关性,多巴胺D2受体拮抗剂对C8胶质细胞促增殖作用有协同性,不同浓度多巴胺对C8胶质细胞中CIB1及LMO3的表达影响不同,具体作用机制还有待进一步研究。
Objective To investigate the effects of Dopamine on the proliferation of C8 glial cells,the possible acceptor pathway and the expressions of CIB1 and LMO3. Methods The effects of Dopamine,Dopamine Agonists and Dopamine Receptor Antagonists on proliferation of C8 glial cells was detected using MTT( thiazolyl blue) assay,and the effects of different concentrations of Dopamine on expressions and locations of LMO3 and CIB1 in C8 glial cells were detected using immunofluorescence method. Results The effects of Dopamine on C8 glial cells obviously related with the its concentration. Low-concentration( 10μmol / L) group had no significant effect on cell growth,middle-concentration( 100 μmol/L) group significantly enhanced cell proliferation( P < 0. 05,P < 0. 01),while high-concentration( 500 μmol/L) group inhibited the cell growth( P < 0. 05). Dopamine-treated group was found more significantly effect on the cell growth than that in D1 agonist group,while the number of C8 glial cells were significant increased by treated with Dopamine and D2 receptor antagonist than by Dopamine( P < 0. 01). The LMO3 expression was significantly increased in low-concentration group( P < 0. 05),while the expression of CIB1 protein was decreased in high-concentration group( P < 0. 05). Conclusion The Dopamine may affect the proliferation of C8 glial cells by Dopamine D2 receptors,and it closely relates to Dopamine concentrations. D2 Dopamine receptor antagonist has the cooperativity with the proliferation of C8 glial cells,and different concentrations of Dopamine can affect expressions of CIB1 and LMO3 in C8 glial cells in different degree.
Objective To investigate the effects of Dopamine on the proliferation of C8 glial cells,the possible acceptor pathway and the expressions of CIB1 and LMO3. Methods The effects of Dopamine,Dopamine Agonists and Dopamine Receptor Antagonists on proliferation of C8 glial cells was detected using MTT( thiazolyl blue) assay,and the effects of different concentrations of Dopamine on expressions and locations of LMO3 and CIB1 in C8 glial cells were detected using immunofluorescence method. Results The effects of Dopamine on C8 glial cells obviously related with the its concentration. Low-concentration( 10μmol / L) group had no significant effect on cell growth,middle-concentration( 100 μmol/L) group significantly enhanced cell proliferation( P < 0. 05,P < 0. 01),while high-concentration( 500 μmol/L) group inhibited the cell growth( P < 0. 05). Dopamine-treated group was found more significantly effect on the cell growth than that in D1 agonist group,while the number of C8 glial cells were significant increased by treated with Dopamine and D2 receptor antagonist than by Dopamine( P < 0. 01). The LMO3 expression was significantly increased in low-concentration group( P < 0. 05),while the expression of CIB1 protein was decreased in high-concentration group( P < 0. 05). Conclusion The Dopamine may affect the proliferation of C8 glial cells by Dopamine D2 receptors,and it closely relates to Dopamine concentrations. D2 Dopamine receptor antagonist has the cooperativity with the proliferation of C8 glial cells,and different concentrations of Dopamine can affect expressions of CIB1 and LMO3 in C8 glial cells in different degree.
引文
[1]于宝成,徐若华,何建政,等.军队男性高龄离退休干部帕金森病患病情况及相关因素调查分析[J].解放军医药杂志,2013,25(8):69-72.
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[3]Miyazaki I,Asanuma M,Diaz-Corrales F J,et al.Direct evidence for expression of dopamine receptors in astrocytes from basal ganglia[J].Brain research,2004,1029(1):120-123.
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[10]Sun W,Guan Q,Wen J,et al.Calcium-and integrinbinding protein-1 is down-regulated in the sperm of patients with oligoasthenozoospermia:CIB1 expression in patients with oligoasthenozoospermia[J].J Assist Reprod Genet,2014,31(5):541-547.
[11]Leisner T M,Moran C,Holly S P,et al.CIB1 prevents nuclear GAPDH accumulation and non-apoptotic tumor cell death via AKT and ERK signaling[J].Oncogene,2013,32(34):4017-4027.
[12]Chan J P,Sieburth D.Localized sphingolipid signaling at presynaptic terminals is regulated by calcium influx and promotes recruitment of priming factors[J].J Neurosci,2012,32(49):17909-17920.
[13]张峪涵,杨磊,黄瑾神.经营养因子用于治疗中枢神经系统疾病[J].中国综合临床,2005,21(3):283-285.
[14]屈赵,张丽,李林,等.不同剂量MOG35-55对EAE小鼠p-JAK1、BDNF表达的影响[J].中国首都医科大学学报,2014,35(6):725-729.
[15]吕彦,何凡,曲方,等.海湾战争综合征模型大鼠海马脑源性神经营养因子及其受体和调节因子的表达[J].解放军预防医学杂志,2013,31(6):490-492.
[16]Khan Z U,Koulen P,Rubinstein M,et al.An astroglialinked dopamine D2-receptor action in prefrontal cortex[J].Proc Natl Acad Sci U S A,2001,98(4):1964-1949.
[17]袁崇刚,薛小琳.多巴胺对PC12细胞的双重影响[J].实验生物学报,2002,35(2):98-102.
[18]Seeman P,Weinshenker D,Quirion R,et al.Dopamine supersensitivity correlates with D2High states,implying many paths to psychosis.Proceedings of the National Academy of Sciences of the United States of America[J].See comment in Pub Med Commons below Proc Natl Acad Sci U S A,2005,102(9):3513-3518.
[19]Zhen X,Torres C,Cai G,et al.Inhibition of protein tyrosine/mitogen-activated protein kinase phosphatase activity is associated with D2 dopamine receptor supersensitivity in a rat model of Parkinson's disease[J].Molecular pharmacology,2002,62(6):1356-1363.
[20]Gerfen C R,Miyachi S,Paletzki R,et al.D1 dopamine receptor supersensitivity in the dopamine-depleted striatum results from a switch in the regulation of ERK1/2/MAP kinase[J].See comment in Pub Med Commons below J Neurosci,2002,22(12):5042-5054.
[21]Espinoza S,Ghisi V,Emanuele M,et al.Postsynaptic D2 dopamine receptor supersensitivity in the striatum of mice lacking TAAR1[J].Neuropharmacology,2015,93:308-313.
[22]Shi J,Cai W,Chen X,et al.Identification of dopamine responsive mRNAs in glial cells by suppression subtractive hybridization[J].Brain research,2001,910(1-2):29-37.
[23]Hui L,Ji C,Hui B,et al.The oncoprotein LMO3 interacts with calcium-and integrin-binding protein CIB1[J].Brain research.2009,1265:24-29.
[2]韩炜,张小青,郭艳丹,等.帕金森病的防治与护理进展[J].解放军预防医学杂志,2015,33(1):100-102.
[3]Miyazaki I,Asanuma M,Diaz-Corrales F J,et al.Direct evidence for expression of dopamine receptors in astrocytes from basal ganglia[J].Brain research,2004,1029(1):120-123.
[4]Hattori A,Luo Y,Umegaki H,et al.Intrastriatal injection of dopamine results in DNA damage and apoptosis in rats[J].Neuroreport,1998,9(11):2569-1572.
[5]Shao W,Zhang S Z,Tang M,et al.Suppression of neuroinflammation by astrocytic dopamine D2 receptors via alpha B-crystallin[J].Nature,2013,494(7435):90-94.
[6]Beardsley P M,Hauser K F.Glial modulators as potential treatments of psychostimulant abuse[J].Advances in pharmacology,2014,69:1-69.
[7]Savarese A,Zou M E,Kharazia V,et al.Increased behavioral responses to ethanol in Lmo3 knockout mice[J].Genes Brain Behav,2014.13(8):777-783.
[8]Watanabe H,Francis J M,Woo M S,et al.Integrated cistromic and expression analysis of amplified NKX2-1 in lung adenocarcinoma identifies LMO3 as a functional transcriptional target[J].Genes Dev,2013,27(2):197-210.
[9]Isogai E,Ohira M,Ozaki T,et al.Oncogenic LMO3 collaborates with HEN2 to enhance neuroblastoma cell growth through transactivation of Mash1[J].PLo S One,2011,6(5):19297.
[10]Sun W,Guan Q,Wen J,et al.Calcium-and integrinbinding protein-1 is down-regulated in the sperm of patients with oligoasthenozoospermia:CIB1 expression in patients with oligoasthenozoospermia[J].J Assist Reprod Genet,2014,31(5):541-547.
[11]Leisner T M,Moran C,Holly S P,et al.CIB1 prevents nuclear GAPDH accumulation and non-apoptotic tumor cell death via AKT and ERK signaling[J].Oncogene,2013,32(34):4017-4027.
[12]Chan J P,Sieburth D.Localized sphingolipid signaling at presynaptic terminals is regulated by calcium influx and promotes recruitment of priming factors[J].J Neurosci,2012,32(49):17909-17920.
[13]张峪涵,杨磊,黄瑾神.经营养因子用于治疗中枢神经系统疾病[J].中国综合临床,2005,21(3):283-285.
[14]屈赵,张丽,李林,等.不同剂量MOG35-55对EAE小鼠p-JAK1、BDNF表达的影响[J].中国首都医科大学学报,2014,35(6):725-729.
[15]吕彦,何凡,曲方,等.海湾战争综合征模型大鼠海马脑源性神经营养因子及其受体和调节因子的表达[J].解放军预防医学杂志,2013,31(6):490-492.
[16]Khan Z U,Koulen P,Rubinstein M,et al.An astroglialinked dopamine D2-receptor action in prefrontal cortex[J].Proc Natl Acad Sci U S A,2001,98(4):1964-1949.
[17]袁崇刚,薛小琳.多巴胺对PC12细胞的双重影响[J].实验生物学报,2002,35(2):98-102.
[18]Seeman P,Weinshenker D,Quirion R,et al.Dopamine supersensitivity correlates with D2High states,implying many paths to psychosis.Proceedings of the National Academy of Sciences of the United States of America[J].See comment in Pub Med Commons below Proc Natl Acad Sci U S A,2005,102(9):3513-3518.
[19]Zhen X,Torres C,Cai G,et al.Inhibition of protein tyrosine/mitogen-activated protein kinase phosphatase activity is associated with D2 dopamine receptor supersensitivity in a rat model of Parkinson's disease[J].Molecular pharmacology,2002,62(6):1356-1363.
[20]Gerfen C R,Miyachi S,Paletzki R,et al.D1 dopamine receptor supersensitivity in the dopamine-depleted striatum results from a switch in the regulation of ERK1/2/MAP kinase[J].See comment in Pub Med Commons below J Neurosci,2002,22(12):5042-5054.
[21]Espinoza S,Ghisi V,Emanuele M,et al.Postsynaptic D2 dopamine receptor supersensitivity in the striatum of mice lacking TAAR1[J].Neuropharmacology,2015,93:308-313.
[22]Shi J,Cai W,Chen X,et al.Identification of dopamine responsive mRNAs in glial cells by suppression subtractive hybridization[J].Brain research,2001,910(1-2):29-37.
[23]Hui L,Ji C,Hui B,et al.The oncoprotein LMO3 interacts with calcium-and integrin-binding protein CIB1[J].Brain research.2009,1265:24-29.