脑缺血/ 再灌注损伤大鼠脑组织Th1/Th2 细胞平衡与PARP-1表达的相关性

神经药理学报 ›› 2015, Vol. 5 ›› Issue (4): 14-21.

脑缺血/ 再灌注损伤大鼠脑组织Th1/Th2 细胞平衡与PARP-1表达的相关性

崔玉环，颜娟，赵宝民，董利平，刘占矿，连晶晶，魏玉磊，郑茂东

1. 河北北方学院附属第一医院老年科，张家口，075000，中国
2. 河北北方学院附属第一医院药剂科，张家口，075000，中国
3. 河北北方学院附属第一医院检验科，张家口，075000，中国
4. 河北北方学院附属第一医院心外科，张家口，075000，中国

出版日期:2015-08-26 发布日期:2016-03-03
通讯作者: 刘占矿，男，主任医师；研究方向：脑血管病研究；Tel：+86-0313-8041513，E-mail：lzk@163.com 董利平，女，主任医师；研究方向：脑血管病研究；Tel：+86-0313-8041513，E-mail：dlp2112@163.com
作者简介:崔玉环，女，主治医师；研究方向：脑血管病研究；E-mail：cuiyuhuan304@163.com
基金资助:
河北省卫生厅科研基金项目（ZL20140035）

Correlation of Th1/Th2 Cells in Balance and PARP-1 Expression in Rat with Cerebral Ischemia/Reperfusion Injury

CUI Yu-huan, YAN Juan, ZHAO Bao-min, DONG Li-ping, LIU Zhan-kuang, LIAN Jing-jing, WEI Yu-lei, ZHENG Mao-dong

1. Department of geriatrics, The First Affiliated Hospital of Hebei North University, Zhangjiakou, 075000, China
2. Department of pharmacy, The First Affiliated Hospital of Hebei North University, Zhangjiakou, 075000, China
3. Department of clinical laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou, 075000, China
4. Department of cardiac surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou, 075000, China

Online:2015-08-26 Published:2016-03-03
Contact: 刘占矿，男，主任医师；研究方向：脑血管病研究；Tel：+86-0313-8041513，E-mail：lzk@163.com 董利平，女，主任医师；研究方向：脑血管病研究；Tel：+86-0313-8041513，E-mail：dlp2112@163.com
About author:崔玉环，女，主治医师；研究方向：脑血管病研究；E-mail：cuiyuhuan304@163.com
Supported by:
河北省卫生厅科研基金项目（ZL20140035）

摘要/Abstract

摘要： 目的：探讨大鼠大脑中动脉阻塞（middle cerebral artery occlusion，MCAO）模型脑组织Th1/Th2细胞平衡与二磷酸腺苷核糖多聚酶-1（Poly（ADP-ribose）polymerase-1，PARP-1）表达是否存在相关性。方法：将健康雄性SD大鼠随机分成正常对照组、假手术组、模型组、PARP-1抑制剂PJ34组，按再灌注不同时间点分为6、12、24、48、72h五个亚组，各组取材前进行神经功能损伤评分，氯化三苯基四氮唑（triphenyltetrazolium chloride，TTC）染色评价脑梗死体积，定量RT-PCR技术检测脑缺血/再灌注损伤病灶中PARP-1 mRNA的表达，Western Blot技术检测PARP-1聚合体蛋白的表达，ELISA技术测定干扰素-γ（interferon-γ，INF-γ）、白介素4（interleukin 4，IL4）水平。结果：与假手术组比，模型组神经功能损伤评分、脑梗死体积、PARP-1表达及炎性反应效应细胞（helper T cell 1，Th1）与辅助性效应细胞（helper T cell 2，Th2）的比值均显著增加（P<0.01）；与模型组比，PJ34组梗死体积明显减小（P<0.01），PARP-1表达、Th1/Th2比值均显著降低（P<0.01）；Th1/Th2与PARP-1水平呈显著正相关（ r=0.984，P<0.05）。结论：MCAO大鼠脑组织Th1 /Th2细胞平衡状态与PARP-1表达存在相关性。

关键词: 二磷酸腺苷核糖多聚酶-1, Th1/Th2比值, 大脑中动脉阻塞

Abstract: Objective: To discuss correlation of Th1/Th2 cells in balance and PARP-1 expression in rat with cerebral ischemia/reperfusion injury. Methods: SD rats were subjected to middle cerebral artery occlusion by suture method, then they were randomly divided into the control group, sham group, model group, PJ34 group, then groups were divided into 5 subgroups according to different reperfusion time points, 6, 12, 24, 48, 72 h, according to the different time points, neurological deficits of each rat were determined infarct volume was analyzed with 2, 3, 5- triphenyltetrazolium chloride (TTC) staining, the protein and mRNA expression of PARP-1 were detected by real-time quantitative PCR and western blot; the level of INF-γ and IL-4 were determined by ELISA methods. Results: compare to sham group, the neurological scores, infarct volume, PARP-1 expression and the Th1/Th2 ratio significantly increased at different time points of reperfusion(P<0.01), and reached a peak by 24h. Compare to model group, neurological scores, infarct volume, PARP-1 expression and the Th1/Th2 ratio significantly reduced in PJ34 group(P<0.01), especially in ischemia reperfusion 24h. PARP-1 expression and the Th1/Th2 ratio had significant positive correlation(r=0.984, P<0.05). Conclusion: Th1/Th2 cells in Balance had related to PARP-1 expression in rat with cerebral ischemia/reperfusion injury.

Key words: Poly（ADP-ribose）polymerase-1(PARP-1), Th1/Th2 ratio, middle cerebral artery occlusion

崔玉环，颜娟，赵宝民，董利平，刘占矿，连晶晶，魏玉磊，郑茂东. 脑缺血/ 再灌注损伤大鼠脑组织Th1/Th2 细胞平衡与PARP-1表达的相关性[J]. 神经药理学报, 2015, 5(4): 14-21.

CUI Yu-huan, YAN Juan, ZHAO Bao-min, DONG Li-ping, LIU Zhan-kuang, LIAN Jing-jing, WEI Yu-lei, ZHENG Mao-dong. Correlation of Th1/Th2 Cells in Balance and PARP-1 Expression in Rat with Cerebral Ischemia/Reperfusion Injury[J]. Acta Neuropharmacologica, 2015, 5(4): 14-21.

参考文献

[1]Ma Ying-xin, Nie Hui; Chen He-yu, et al. NAD(+)/NADH metabolism and NAD(+)- dependent enzymes in cell death and ischemic brain injury: current advances and therapeutic implications[J]. Curr Med Chem, 2015, 22(10): 1239-1247.

[2]Gerace E, Pellegrini-Giampietro D E, Moroni F, et al. Poly(ADP-Ribose) polymerase 1 (PARP-1) activation and Ca(2+) permeable alpha-amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) channels in post-ischemic brain damage: new therapeutic opportunities?[J]. CNS Neurol Disord Drug Targets, 2015, 14(5): 636-646.

[3]Xiong Xiao-xing, Barreto George, Xu Li-jun, et al. Increased brain injury and worsened neurological outcome in interleukin-4 knockout mice after transient focal cerebral ischemia[J]. Stroke, 2011, 42(7): 2026-2032.

[4]Xie Luo-kun, Sun Fen, Wang Ji-xian, et al. mTOR signaling inhibition modulates macrophage/microglia-mediated neuroinflammation and secondary injury via regulatory T cells after focal ischemia[J]. J Immunol, 2014, 192(12): 6009-6019.

[5]Yun Wu, Qing-Cheng Liang, Lei Yang, et al. Mucosal tolerance to E-selectin provides protection against cerebral ischemia-reperfusion injury in rats[J]. J Neuroimmunol, 2008, 205(1-2): 73-79.

[6]LuoXin, W Lee Kraus. On PAR with PARP: cellular stress signaling through poly(ADP-ribose) and PARP-1[J]. Genes Dev, 2012, 26(5): 417-432.

[7]Sabrina Giacoppo, Maria Galuppo, Renato Iori, et al. (RS)-glucoraphanin purified from Tuscan black kale and bioactivated with myrosinase enzyme protects against cerebral ischemia/reperfusion injury in rats[J]. Fitoterapia, 2014, 99166-99177.

[8]Rosaria Greco, Cristina Tassorelli, Antonina Stefania Mangione, et al. Neuroprotection by the PARP inhibitor PJ34 modulates cerebral and circulating RAGE levels in rats exposed to focal brain ischemia[J]. Eur J Pharmacol, 2014, 74491-74497.

[9]Muralidhar L Hegde, Anil K Mantha, Tapas K Hazra, et al. Oxidative genome damage and its repair: implications in aging and neurodegenerative diseases[J]. Mech Ageing Dev, 2012, 133: 157 168.

[10]Masahito Kawabori, Midori A Yenari . Inflammatory responses in brain ischemia[J]. Curr Med Chem, 2015, 22(10) : 1258-1277.

[11]Femando P Rodrigues, Cezar R Pestana, Ana C M Polizello, et al. Release of NO from a nitrosyl ruthenium complex through oxidation of mitochondrial NADH and effects on mitochondria[J]. Nitric Oxide, 2012, 26(13): 174 181.

[12]Zhang Rui-xue, Tang Shi, Huang Wei-wei, et al. Protection of the brain following cerebral ischemia through the attenuation of PARP-1-induced neurovascular unit damage in rats[J]. Brain Res, 2015, 1624: 9-18.

[13]Marianne Haddad, Virginie Beray-Berthat, Berard Coqueran, et al. Combined therapy with PJ34, a poly(ADP-ribose)polymerase inhibitor, reduces tissue plasminogen activator- induced hemorrhagic transformations in cerebral ischemia in mice[J]. Fundam Clin Pharmacol, 2013, 27(4): 393-401.

[14]M Haddad, H RhinnC, Bloquel, et al. Anti-inflammatory effects of PJ34, a poly (ADP-ribose) polymerase inhibitor, in transient focal cerebral ischemia in mice[J]. British J Pharmacology, 2006, 149(1): 23–30.

[15]Egi Y, Matsuura S, Maruyama T, et al. Neuroprotective effects of a novel water-soluble poly(ADP-ribose) polymerase-1 inhibitor, MP-124, in in vitro and in vivo models of cerebral ischemia[J]. Brain Res, 2011, 1389: 169-176.

[16]Tiina Kauppinen, Sang Won Suh, Ari E Berman, et al. Inhibition of poly(ADP-ribose) polymerase suppresses inflammation and promotes recovery after ischemic injury[J]. J Cereb Blood Flow Metab, 2009, 29(4): 820-829.

[17]Slava Rom, Viviana Zuluaga-Ramirez, Holly Dykstra, et al. Poly(ADP-ribose) polymerase-1 inhibition in brain endothelium protects the blood-brain barrier under physiologic and neuroinflammatory conditions[J]. J Cereb Blood Flow Metab, 2015, 35(1): 28-36.

[18]Nakajima H, Kubo T, Ihara H, et al. Nuclear-translocated glyceraldehyde-3- phosphate dehydrogenase promotes poly(ADP-ribose) polymerase-1 activation during oxidative/nitrosative stress in stroke[J]. J Biol Chem, 2015, 290(23): 14493-14503.

[19]Federical Laudisi, Manolo Sambucci, Claudio Pioli. Poly (ADP-ribose) polymerase-1 (PARP-1) as immune regulator[J]. Endocr Metab Immune Disord Drug Targets, 2011, 11(4): 326-333.