Janus激酶/信号传导和转录激活蛋白3通路阻断对新生大鼠缺氧缺血性脑病的影响
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摘要
目的探讨Janus激酶/信号传导和转录激活蛋白3(JAK/STAT3)通路阻断对新生大鼠缺氧缺血性脑病(HIE)的影响,为HIE的基因治疗提供依据。方法将70只Wistar新生大鼠随机分为对照组(10只)、HIE组(30只)和干预组(30只)。HIE组和干预组新生大鼠制备HIE模型,对照组新生大鼠不制备HIE模型。干预组新生大鼠于造模前10 min腹腔注射JAK/STAT3信号通路阻断剂AG490 3 mg·kg~(-1)。HIE组和干预组新生大鼠分别于造模后6、48、72 h处死(每组每个时间点处死10只),对照组新生大鼠于48 h时全部处死,取大脑海马组织,采用苏木精-伊红(HE)染色观察新生大鼠海马组织病理学改变,采用实时荧光定量聚合酶链反应检测大鼠海马组织中微小RNA-21(miR-21)和STAT3 mRNA的表达。结果 HE染色显示,对照组新生大鼠海马组织中神经元和小胶质细胞的大小、形态均正常,未见细胞变性及水肿现象;HIE组新生大鼠海马组织中神经元体积增大,中度水肿,着色不均;干预组新生大鼠海马组织中神经元体积增大,中、重度水肿,着色不均,可见细胞核固缩,并见小胶质细胞轻度增生,间质水肿。造模后6、48、72 h,HIE组和干预组大鼠海马组织中miR-21、STAT3 mRNA相对表达量显著高于对照组(P<0.05),干预组大鼠海马组织中miR-21、STAT3 mRNA相对表达量显著低于HIE组(P<0.05)。HIE组和干预组大鼠造模后48 h时海马组织中miR-21、STAT3 mRNA相对表达量显著高于造模后6、72 h(P<0.05),HIE组和干预组大鼠造模后6 h与造模后72 h时海马组织中miR-21、STAT3 mRNA相对表达量比较差异无统计学意义(P>0.05)。结论 HIE新生大鼠海马组织中STAT3 mRNA、miR-21表达升高可能对神经细胞起到应激性保护作用,这一作用可以被JAK/STAT3信号传导通路阻断剂AG490阻断。
Objective To investigate the effect of Janus kinase/signal transducers and activators of transcription 3( JAK/STAT3) pathway inhibition on hypoxic ischemic encephalopathy( HIE) in neonatal rats,and to provide evidence for gene therapy of HIE. Methods Seventy Wistar neonatal rats were randomly divided into control group( n = 10),HIE group( n = 30) and intervention group( n = 30). The HIE models were established in neonatal rats of the HIE group and the intervention group,but the HIE model was not prepared in neonatal rats of the control group. The neonatal rats in the intervention group were treated with JAK/STAT3 signaling pathway blocker AG490( 3 mg·kg~(-1)) by intraperitoneal injection at 10 minutes before modeling. The neonatal rats in the HIE group and the intervention group were executed at 6,48 and 72 hours after modeling( 10 rats in each group at each time point),and the neonatal rats in the control group were all executed at48 hours after modeling,and the hippocampal tissues were obtained. The histopathological changes of hippocampal tissues of neonatal rats were observed by hematoxylin-eosin( HE) staining. The expression of microRNA-21( miR-21) and STAT3 in hippocampal tissues of rats was detected by real-time fluorescence quantitative polymerase chain reaction. Results HE staining showed that the size and morphology of neurons and microglia in hippocampus of neonatal rats in the control group were normal,and no cell degeneration and edema were observed; the neurons in hippocampus of neonatal rats in the HIE group were enlarged,moderately edematous and unevenly colored; the morphological changes of hippocampal tissues of neonatal rats in the intervention group included neuron enlargement,moderate to severe edema,uneven staining,karyopyknosis,slight proliferation of microglia and interstitial edema. At 6,48 and 72 hours after modeling,the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats in the HIE group and intervention group was significantly higher than that in the control group( P < 0. 05),and the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats in the intervention group was significantly lower than that in the HIE group( P < 0. 05). The relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats at 48 hours after modeling was significantly higher than that at 72 hours after modeling in the HIE group and intervention group( P < 0. 05). There was no significant difference in the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats between the time points of 6 hours and72 hours after modeling in the HIE group and intervention group( P > 0. 05). Conclusion The up-regulation of the expression of STAT3 mRNA and miR-21 in the hippocampal tissues of neonatal HIE rats may play a stress protective role on nerve cells,which can be blocked by JAK/STAT3 signal transduction pathway blocker AG490.
Objective To investigate the effect of Janus kinase/signal transducers and activators of transcription 3( JAK/STAT3) pathway inhibition on hypoxic ischemic encephalopathy( HIE) in neonatal rats,and to provide evidence for gene therapy of HIE. Methods Seventy Wistar neonatal rats were randomly divided into control group( n = 10),HIE group( n = 30) and intervention group( n = 30). The HIE models were established in neonatal rats of the HIE group and the intervention group,but the HIE model was not prepared in neonatal rats of the control group. The neonatal rats in the intervention group were treated with JAK/STAT3 signaling pathway blocker AG490( 3 mg·kg~(-1)) by intraperitoneal injection at 10 minutes before modeling. The neonatal rats in the HIE group and the intervention group were executed at 6,48 and 72 hours after modeling( 10 rats in each group at each time point),and the neonatal rats in the control group were all executed at48 hours after modeling,and the hippocampal tissues were obtained. The histopathological changes of hippocampal tissues of neonatal rats were observed by hematoxylin-eosin( HE) staining. The expression of microRNA-21( miR-21) and STAT3 in hippocampal tissues of rats was detected by real-time fluorescence quantitative polymerase chain reaction. Results HE staining showed that the size and morphology of neurons and microglia in hippocampus of neonatal rats in the control group were normal,and no cell degeneration and edema were observed; the neurons in hippocampus of neonatal rats in the HIE group were enlarged,moderately edematous and unevenly colored; the morphological changes of hippocampal tissues of neonatal rats in the intervention group included neuron enlargement,moderate to severe edema,uneven staining,karyopyknosis,slight proliferation of microglia and interstitial edema. At 6,48 and 72 hours after modeling,the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats in the HIE group and intervention group was significantly higher than that in the control group( P < 0. 05),and the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats in the intervention group was significantly lower than that in the HIE group( P < 0. 05). The relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats at 48 hours after modeling was significantly higher than that at 72 hours after modeling in the HIE group and intervention group( P < 0. 05). There was no significant difference in the relative expression of miR-21 and STAT3 mRNA in hippocampal tissues of neonatal rats between the time points of 6 hours and72 hours after modeling in the HIE group and intervention group( P > 0. 05). Conclusion The up-regulation of the expression of STAT3 mRNA and miR-21 in the hippocampal tissues of neonatal HIE rats may play a stress protective role on nerve cells,which can be blocked by JAK/STAT3 signal transduction pathway blocker AG490.
引文
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[13]王玉,席乐峰,杨廷桐.C-myc与肺鳞癌中miRNA-21表达的关系[J].解剖学杂志,2013,36(4):758-761.
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[16]孔令平,刘爱芹,周旋,等.STAT-3抑制剂WP1066增强顺铂对口腔鳞状细胞癌侵袭能力抑制作用的体外研究[J].天津医药,2016,44(1):38-42.
[2]何畏,方伟蓉,李运曼.JAK/STAT通路与脑缺血损伤研究进展[J].药学与临床研究,2015,23(2):164-167.
[3]王来栓,邹亮燕.间充质干细胞移植治疗新生儿缺氧缺血性脑病研究进展[J].中华实用儿科临床杂志,2018,33(2):81-85.
[4]陈惠军,杨廷桐.新生儿缺氧缺血性脑病血清中miR-21调控HIF-1a表达及临床意义[J].中国儿童保健杂志,2015,23(1):32-35.
[5]LIN Y,CAI B,XUE X H,et al.TAT-mediated delivery of neuroglobin attenuates apoptosis induced by oxygen-glucose deprivation via the Jak2/Stat3 pathway in vitro[J].Neurol Res,2015,37(6):531-538.
[6]LIU R,XU N,YI W,et al.Electroacupuncture effects on cortical neurons,as well as Janus kinase 2-signal transducer and activator of transcription 3 signal transduction pathway,in a rat model of cerebral ischemia[J].Neural Regen Res,2012,7(6):457-462.
[7]孙向峰,侯梅,苑爱云,等.丰富环境对缺氧缺血性脑损伤新生大鼠学习记忆、海马区脑源性神经生长因子和突触素蛋白表达的影响[J].中华实用儿科临床杂志,2017,32(4):296-299.
[8]LUO Y,YANG Z,SU L,et al.Non-CSCs nourish CSC sthrough interleukin-17 E-mediated activation of NF-κB and JAK/STAT3signaling in human hepatocellular carcinoma[J].Cancer Lett,2016,375(2):390-399.
[9]庞炜,曹帅帅,李树祎,等.新生儿缺氧缺血脑损伤大鼠模型的制备[J].中国比较医学杂志,2016,26(6):61-66.
[10]石计朋,栗延伟,郭丽娟,等.ω-3多不饱和脂肪酸对脂多糖所致脑损伤新生大鼠海马组织中氧化应激产物和细胞凋亡的影响[J].新乡医学院学报,2018,35(6):455-458.
[11]SUN Y,NADAL-VICENS M,MISONO S,et al.Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms[J].Cell,2001,104(3):365-376.
[12]万冬,杨廷桐,秦玉凤,等.miRNA-21/PDCD4环路在卵巢癌组织中的表达[J].肿瘤防治研究,2013,40(9):869-872.
[13]王玉,席乐峰,杨廷桐.C-myc与肺鳞癌中miRNA-21表达的关系[J].解剖学杂志,2013,36(4):758-761.
[14]ROZOVSKI U,CALIN G A,SETOYAMA T,et al.Signal tranducer and actiwator of transcription(STAT)-3 regulates microRNA gene expression in chronic lymphocytic leukemia cells[J].Mol Cancer,2013,50(12):1476-1481.
[15]LI Y,WU R,LIU Z,et al.Enforced expression of microRNA-21influences the replication of varicella-zoster virus by triggering signal transducer and activator of transcription 3[J].Exp Ther Med,2014,7(5):1291-1296.
[16]孔令平,刘爱芹,周旋,等.STAT-3抑制剂WP1066增强顺铂对口腔鳞状细胞癌侵袭能力抑制作用的体外研究[J].天津医药,2016,44(1):38-42.
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