JAK2/STAT3通路介导热休克蛋白70对脑出血的神经保护作用
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摘要
目的观察蛋白酪氨酸激酶2/信号转导和转录激活因子3(janus kinase 2-signal transducer and activator of transcription 3,JAK2/STAT3)信号通路特异性拮抗剂α-氰基-(3,4-羟基)N-苄苯乙烯胺〔alpha cyano-(3-hydroxy)N-styrene benzyl amine,AG-490〕对脑出血大鼠脑组织热休克蛋白70(heat shock protein 70,HSP70)、磷酸化蛋白酪氨酸激酶2(phosphorylated janus kinase 2,P-JAK2)、磷酸化信号转导和转录激活因子3(phosphorylated signal transducer and activator of transcription 3,P-STAT3)表达的影响,并分析HSP70与PJAK2、P-STAT3的相关性,探讨JAK2/STAT3信号通路是否介导HSP70对大鼠脑出血后的神经保护作用。方法将健康雄性Sprague-Dawley(SD)大鼠100只随机分为正常组、假手术组、脑出血和AG-490组;采用大鼠自体血注入法建立脑出血模型;各组按造模成功后时间分为6h、24h、48h、72h、7d共5个亚组,对各组大鼠进行神经功能缺损评分(neurological severity scores,NSS);免疫组化染色检测血肿周围HSP70表达,采用Western blot检测大鼠血肿周围脑组织P-JAK2、P-STAT3的表达。结果脑出血组和AG-490组HSP70、P-JAK2、PSTAT表达均高于假手术组和正常组(P<0.05),并于造模后48h达高峰(P<0.05);AG-490组HSP70、PJAK2、P-STAT表达较脑出血组低(P<0.05);脑出血组HSP70表达与NSS评分呈负相关(r=-0.511,P<0.05),HSP70表达与P-JAK2、P-STAT3的表达呈正相关(r=0.790,P<0.05;r=0.447,P<0.05)。结论 JAK2/STAT3信号通路可能通过上调HSP70的表达进而对大鼠脑出血后起神经保护作用。
Objective To investigate the effect of Alpha cyano-(3-hydroxy)N-styrene benzyl amine(AG-490)[the janus kinase 2-signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathways specific antagonist]on the expression of heat shock protein 70(HSP70),phosphorylated janus kinase 2(P-JAK2)and activator of transcription 3(P-STAT3)in cerebral hemorrhage rat models,and the correlation between the JAK2/STAT3 signaling pathway and the expression of HSP70.To investigate the role of JAK2/STAT3 signal pathway and HSP70 in neuroprotective effects of cerebral hemorrhage rats.Methods One hundred healthy male Sprague-Dawley rats were randomly divided into four groups:the normal group,the sham operation group,the cerebral hemorrhage group and the AG-490 group.The autologous arterial blood was stereotaxically injected to the right caudate nucleus to establish cerebral hemorrhage rat models.According to the different sacrificed time,every group was divided into 5subgroups:6h,24 h,48h,72 h,7d.The rats were evaluated by the neurological deficit scores(Neurological Severity Scores,NSS),the expressions of HSP70,PJAK2,and P-STAT3 were detected by immunostaining and Western blot using rat brain tissue around the hematoma.Results HSP70,P-JAK2 and P-STAT expressions in the cerebral hemorrhage group and the AG-490 group were higher than those of the sham group and the normal group(P<0.05),and 48 hafter modeling their expressions reached a peak(P<0.05).HSP70,P-JAK2 and P-STAT expressions in the AG-490 group were lower than those of the cerebral hemorrhage group(P<0.05).NSS and HSP70 expression were negatively correlated(r=-0.511,P<0.05),HSP70 expression was positively correlated with P-JAK2 and P-STAT3expression(r=0.790,P<0.05;r=0.447,P<0.05)in the cerebral hemorrhage group.Conclusions JAK2/STAT3 signaling pathway may play a role in neuroprotection after cerebral hemorrhage by upregulating the expression of HSP70.
Objective To investigate the effect of Alpha cyano-(3-hydroxy)N-styrene benzyl amine(AG-490)[the janus kinase 2-signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathways specific antagonist]on the expression of heat shock protein 70(HSP70),phosphorylated janus kinase 2(P-JAK2)and activator of transcription 3(P-STAT3)in cerebral hemorrhage rat models,and the correlation between the JAK2/STAT3 signaling pathway and the expression of HSP70.To investigate the role of JAK2/STAT3 signal pathway and HSP70 in neuroprotective effects of cerebral hemorrhage rats.Methods One hundred healthy male Sprague-Dawley rats were randomly divided into four groups:the normal group,the sham operation group,the cerebral hemorrhage group and the AG-490 group.The autologous arterial blood was stereotaxically injected to the right caudate nucleus to establish cerebral hemorrhage rat models.According to the different sacrificed time,every group was divided into 5subgroups:6h,24 h,48h,72 h,7d.The rats were evaluated by the neurological deficit scores(Neurological Severity Scores,NSS),the expressions of HSP70,PJAK2,and P-STAT3 were detected by immunostaining and Western blot using rat brain tissue around the hematoma.Results HSP70,P-JAK2 and P-STAT expressions in the cerebral hemorrhage group and the AG-490 group were higher than those of the sham group and the normal group(P<0.05),and 48 hafter modeling their expressions reached a peak(P<0.05).HSP70,P-JAK2 and P-STAT expressions in the AG-490 group were lower than those of the cerebral hemorrhage group(P<0.05).NSS and HSP70 expression were negatively correlated(r=-0.511,P<0.05),HSP70 expression was positively correlated with P-JAK2 and P-STAT3expression(r=0.790,P<0.05;r=0.447,P<0.05)in the cerebral hemorrhage group.Conclusions JAK2/STAT3 signaling pathway may play a role in neuroprotection after cerebral hemorrhage by upregulating the expression of HSP70.
引文
[1]徐海发,蒋晶晶,马静萍.重组人粒细胞集落刺激因子对大鼠脑缺血再灌注损伤后p-JAK2及p-STAT3信号通路的影响[J].中国神经免疫学和神经病学杂志,2014,21(4):273-277.
[2]Chen G,Zhang S,Shi J,et al.Effects of recombinant human erythropoietin(rhEPO)on JAK2/STAT3pathway and endothelial apoptosis in the rabbit basilar artery after subarachnoid hemorrhage[J].Cytokine,2009,45(3):162-168.
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[4]Madamanchi NR,Li S,Patterson C,et al.Reactiveoxy genspecies regulate heat-shock protein 70via the JAK/STAT pathway[J].Arterioscl Throm Vasc,2001,21(3):321-326.
[5]李慧,娄季宇,杨霄鹏.大鼠脑出血模型制作技巧[J].中国实用神经病杂志,2007,10(2):152-153.
[6]Long EZ,Weinstein PR,Carlson S,et al.Reversible Middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[7]Garcia JH,Wagner S,Liu KF,et al.Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rat statistical validation[J].Stroke,1995,26(4):627-635.
[8]Morales JK,Falanga YT,Depcrynski A,et al.Mast cell homeostasis and the JAK-STAT pathway[J].Genes Immun,2010,11(8):599-608.
[9]Lai SY,Johnson FM.Defining the role of the JAK-STAT pathway in head and neck and thoracic malignancies:implications for future therapeutic approaches[J].Drug Resist Updat,2010,13(3):67-78.
[10]Nomura N,Nunes P,Bouley R,et al.High-throughput chemical screening identifies AG-490as a stimulator of aquaporin 2 membrane expression and urine concentration[J].Am J Physiol Cell Physiol,2014,307(7):597-605.
[11]张合鹏,杜爱玲,李磊,等.AG490对大鼠脑损伤后血-脑屏障通透性及白细胞介素6和肿瘤坏死因子α表达的影响[J].中国脑血管病杂志,2015(12),3:134-139.
[12]Choi JI,Kim SD,Kim SH,et al.Semi-quantitative analyses of hippocampal heat shock protein-70expression based on the duration of ischemia and the volume of cerebral infarction in mice[J].J Korean Neurosurg Soc,2014,55(6):307-312.
[13]Gómez-Choco M,Doucerain C,Urra X,et al.Presence of heat shock protein 70in secondary lymphoid tissue correlates with stroke prognosis[J].J Neuroimmunol,2014,15(2):67-74.
[14]Kitamura C,Ogawa Y,Nishihara T,et al.Transient colocalization of c-Jun N-terminal kinase and c-Jun with heat shock protein 70in pulp cells during apoptosis[J].J Dent Res,2003,82(2):91-95.
[15]刘胜中,甘崇志,杨双强.JAK2/STAT3通过上调HSP70蛋白介导TTX心肌保护作用[J].西部医学,2011,23(3):415-418.
[2]Chen G,Zhang S,Shi J,et al.Effects of recombinant human erythropoietin(rhEPO)on JAK2/STAT3pathway and endothelial apoptosis in the rabbit basilar artery after subarachnoid hemorrhage[J].Cytokine,2009,45(3):162-168.
[3]董梅,辛立建,陈丽萍,等.SOCS1-JAK2-STAT3信号通路在C57BL-6小鼠实验性自身免疫性脑脊髓炎模型中作用机制探讨[J].中国免疫学杂志,2014,7(4):459-463.
[4]Madamanchi NR,Li S,Patterson C,et al.Reactiveoxy genspecies regulate heat-shock protein 70via the JAK/STAT pathway[J].Arterioscl Throm Vasc,2001,21(3):321-326.
[5]李慧,娄季宇,杨霄鹏.大鼠脑出血模型制作技巧[J].中国实用神经病杂志,2007,10(2):152-153.
[6]Long EZ,Weinstein PR,Carlson S,et al.Reversible Middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[7]Garcia JH,Wagner S,Liu KF,et al.Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rat statistical validation[J].Stroke,1995,26(4):627-635.
[8]Morales JK,Falanga YT,Depcrynski A,et al.Mast cell homeostasis and the JAK-STAT pathway[J].Genes Immun,2010,11(8):599-608.
[9]Lai SY,Johnson FM.Defining the role of the JAK-STAT pathway in head and neck and thoracic malignancies:implications for future therapeutic approaches[J].Drug Resist Updat,2010,13(3):67-78.
[10]Nomura N,Nunes P,Bouley R,et al.High-throughput chemical screening identifies AG-490as a stimulator of aquaporin 2 membrane expression and urine concentration[J].Am J Physiol Cell Physiol,2014,307(7):597-605.
[11]张合鹏,杜爱玲,李磊,等.AG490对大鼠脑损伤后血-脑屏障通透性及白细胞介素6和肿瘤坏死因子α表达的影响[J].中国脑血管病杂志,2015(12),3:134-139.
[12]Choi JI,Kim SD,Kim SH,et al.Semi-quantitative analyses of hippocampal heat shock protein-70expression based on the duration of ischemia and the volume of cerebral infarction in mice[J].J Korean Neurosurg Soc,2014,55(6):307-312.
[13]Gómez-Choco M,Doucerain C,Urra X,et al.Presence of heat shock protein 70in secondary lymphoid tissue correlates with stroke prognosis[J].J Neuroimmunol,2014,15(2):67-74.
[14]Kitamura C,Ogawa Y,Nishihara T,et al.Transient colocalization of c-Jun N-terminal kinase and c-Jun with heat shock protein 70in pulp cells during apoptosis[J].J Dent Res,2003,82(2):91-95.
[15]刘胜中,甘崇志,杨双强.JAK2/STAT3通过上调HSP70蛋白介导TTX心肌保护作用[J].西部医学,2011,23(3):415-418.