孕酮在脑缺血大鼠TNF-α和IL-6引起脑损伤中的作用
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摘要
目的观察孕酮在炎症因子TNF-α和IL-6引起新生鼠缺氧缺血性脑损伤中的作用,进一步探讨孕酮神经保护作用的分子机制。方法 96只7日龄新生Wistar大鼠随机分成4组,正常对照组、假手术组、缺氧缺血组、药物预防组。缺氧缺血组和药物预防组动物先行左侧颈总动脉结扎术,然后将动物置于37℃恒温的密闭容器中,以1.5L/min的速度吸入80ml/L氧气和920ml/L氮气混合气体2.5h建立缺氧缺血脑病动物模型。药物预防组动物于建立模型前30min按8mg/kg腹腔注射0.5g/L孕酮溶液。采用ELISA法检测脑组织中TNF-α和IL-6含量的变化,RT-PCR检测TNF-α和IL-6 mRNA的表达。结果缺氧缺血组脑组织中炎症因子TNF-α和IL-6含量在缺氧缺血后6h、24h、48h、72h明显高于对照组和假手术组大鼠(P<0.05),且在缺氧后24h升到最高点,以后逐渐下降,至7d两者差异不显著,药物预防组在缺氧缺血后各时间点均低于缺氧缺血组(P<0.05)。且缺氧24h后缺氧缺血组脑组织TNF-α、IL-6 mRNA的表达明显高于正常对照组和假手术组,药物预防组mRNA的表达明显低于缺氧缺血组(P<0.05)。结论孕酮可以保护新生鼠缺氧缺血后引起的脑损伤,其作用机制与抑制炎症因子TNF-α和IL-6 mRNA的表达以及抑制TNF-α和IL-6的生成有关。
Objective To study the role of progesterone in brain damage with ischemia rat caused by TNF-α and IL-6,and discuss the protective molecular mechanism of progesterone.Methods 96 7-day-old neonatal rats were randomly divided into four groups:normal group,sham-operated group,hypoxic-ischemic group and pretreatment groups.Rats in hypoxic-ischemic group and pretreatment groups were subjected to left common carotid artery ligation,then were exposed to 80ml/L oxygen and 920ml/L nitrogen gas in 37℃ closed container for up to 2.5h to establish HIE model.Progesterone was injected intraperitoneally into the rats of pretreatment groups rats at 30 minutes before hypoxia,The contents of TNF-α、IL-6 was measured by ELISA and the expression of TNF-α、IL-6 mRNA was analyzed by RT-PCR.Results The contents of TNF-α and IL-6 in brain tissue in hypoxic-ischemic group were significantly higher than those in normal group and sham-operated group at 6h、24h、48h、72h after hypoxia respectively(P<0.05),and riseds to the highest point at 24h,then gradually reduced,dropped to normal level at 7d.The contents of TNF-α and IL-6 were significantly lower in pretreatment group than that in the hypoxic-ischemic group.And the expression of TNF-α、IL-6 in hypoxic-ischemic group were significantly higher than that in the normal and sham-operated groups at 24h after hypoxia.In pretreatment groups,The expression of TNF-α、IL-6 were significantly lower than that in hypoxic-ischemic group(P<0.05).Conclusion Progesterone exerts neuroprotective effect on hypoxic-ischemic encephalopathy-induced brain damage,the action mechanism was related with down-regulate the expression of TNF-α、IL-6,caused lessitter production of TNF-α and、IL-6 produced.
Objective To study the role of progesterone in brain damage with ischemia rat caused by TNF-α and IL-6,and discuss the protective molecular mechanism of progesterone.Methods 96 7-day-old neonatal rats were randomly divided into four groups:normal group,sham-operated group,hypoxic-ischemic group and pretreatment groups.Rats in hypoxic-ischemic group and pretreatment groups were subjected to left common carotid artery ligation,then were exposed to 80ml/L oxygen and 920ml/L nitrogen gas in 37℃ closed container for up to 2.5h to establish HIE model.Progesterone was injected intraperitoneally into the rats of pretreatment groups rats at 30 minutes before hypoxia,The contents of TNF-α、IL-6 was measured by ELISA and the expression of TNF-α、IL-6 mRNA was analyzed by RT-PCR.Results The contents of TNF-α and IL-6 in brain tissue in hypoxic-ischemic group were significantly higher than those in normal group and sham-operated group at 6h、24h、48h、72h after hypoxia respectively(P<0.05),and riseds to the highest point at 24h,then gradually reduced,dropped to normal level at 7d.The contents of TNF-α and IL-6 were significantly lower in pretreatment group than that in the hypoxic-ischemic group.And the expression of TNF-α、IL-6 in hypoxic-ischemic group were significantly higher than that in the normal and sham-operated groups at 24h after hypoxia.In pretreatment groups,The expression of TNF-α、IL-6 were significantly lower than that in hypoxic-ischemic group(P<0.05).Conclusion Progesterone exerts neuroprotective effect on hypoxic-ischemic encephalopathy-induced brain damage,the action mechanism was related with down-regulate the expression of TNF-α、IL-6,caused lessitter production of TNF-α and、IL-6 produced.
引文
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[11]李新娟,李爽,李东亮,等.孕酮对缺氧缺血性脑损伤新生大鼠血脑屏障通透性及脑水肿的影响[J].中国药理学通报,2011,27(3):364-367.
[12]Li S,Xu C Y,Jiang H B,et al.Effect of progesterone on the expressionof superoxide dismutase and matrix metalloproteinase-3 in the braintissues of neonatal rats with hypoxic-ischemia brain damage[J].J Ap-pl Clin Pediatr,2010,25(14):1065-1067.
[13]Oshima T,Lee S,Sato A,et al.TNF-alpha contributes to axonal sprou-ting and functional recovery following traumatic brain injury[J].BrainRes,2009,1290(1):102-104.
[14]Aly H,Khashaba MT,El-Ayouty M,et al.IL-1beta,IL-6 and TNF-al-pha and outcomes of neonatal hypoxic ischemic encephalopathy[J].Brain Dev,2006,28(3):178-182.
[15]Folkersma H,Breve JJ,Tilders FJ,et al.Cerebral microdialysis of in-terleukin(IL)-1beta and IL-6:extraction efficiency and production inthe acute phase after severe traumatic brain injury in rats[J].ActaNeurochir,2008,150(12):1277-1279.
[2]卢先锋,秦琴,杨晓丽.新生大鼠缺氧缺血性脑损伤热休克蛋白70合成与TNF-α的关系[J].临床儿科杂志,2010,28(9):864-867.
[3]Sairanen T,Carpen O,Karjalainen-Lindsberg ML,et al.Evolution ofcerebral tumor necrosis factor-production during human ischemic stroke[J].Stroke,2001,32:1750-1758.
[4]车玉琴,任玉峰,高杰,等.IGF-Ⅱ对大鼠脑缺血/再灌注损伤后海马TNF-α含量的影响[J].中国应用生理学杂志,2009,25(2):249-250.
[5]连俊兰,余勤,王艳.幼年大鼠缺氧缺血性脑病动物模型的研究[J].现代中西医结合杂志,2007,16(27):3947-3950.
[6]莫国梁.新生儿缺氧缺血性脑病动物模型的建立[J].儿科药学杂志,2008,14(2):58-60.
[7]Wright DW,Hoffman SW,Virmani S,et al.Effects of medroxyprogester-one acetate on cerebral oedema and spatial learning performance aftertraumatic brain injury in rats[J].Brain Inj,2008,22(2):107-113.
[8]Wang J,Jiang C,Liu C,et al.Neuroprotective effects of progesteronefollowing stroke in aged rats[J].Behav Brain Res,2010,209(1):119-122.
[9]Brinton RD,Thompson RF,Foy M R,et al.Progesterone receptors:form and function in brain[J].Front Neuroendocrinol,2008,29(2):313-339.
[10]Micevych P,Soma KK,Sinchak K.Neuroprogesterone:keyto estrogenpositive feedback[J].Brain Res Rev,2008,57(2):470-480.
[11]李新娟,李爽,李东亮,等.孕酮对缺氧缺血性脑损伤新生大鼠血脑屏障通透性及脑水肿的影响[J].中国药理学通报,2011,27(3):364-367.
[12]Li S,Xu C Y,Jiang H B,et al.Effect of progesterone on the expressionof superoxide dismutase and matrix metalloproteinase-3 in the braintissues of neonatal rats with hypoxic-ischemia brain damage[J].J Ap-pl Clin Pediatr,2010,25(14):1065-1067.
[13]Oshima T,Lee S,Sato A,et al.TNF-alpha contributes to axonal sprou-ting and functional recovery following traumatic brain injury[J].BrainRes,2009,1290(1):102-104.
[14]Aly H,Khashaba MT,El-Ayouty M,et al.IL-1beta,IL-6 and TNF-al-pha and outcomes of neonatal hypoxic ischemic encephalopathy[J].Brain Dev,2006,28(3):178-182.
[15]Folkersma H,Breve JJ,Tilders FJ,et al.Cerebral microdialysis of in-terleukin(IL)-1beta and IL-6:extraction efficiency and production inthe acute phase after severe traumatic brain injury in rats[J].ActaNeurochir,2008,150(12):1277-1279.