通腑平喘方对脓毒症急性肺损伤大鼠Keap1-Nrf2/ARE信号通路的影响
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摘要
目的观察通腑平喘方对脓毒症急性肺损伤大鼠核因子E2相关因子(Nrf2)表达的影响,探讨通腑平喘方对脓毒症急性肺损伤抗氧化应激的作用机制。方法采用CLP制作脓毒症模型,将40只成年雄性SD大鼠随机分为正常组、中药组、模型组及假手术组4组,每组10只。正常组于6 h、12 h后予以0.9%氯化钠注射液灌胃。假手术组开腹后将盲肠取出后再回纳到腹腔,术后6 h、12 h予以0.9%氯化钠注射液灌胃。模型组CLP术后立即予以0.9%氯化钠注射液灌胃,术后6 h、12 h予以0.9%氯化钠注射液灌胃。中药组CLP术后立即予以通腑平喘方灌胃,术后6 h、12 h再次予以通腑平喘方灌胃。以上各组均在24 h后麻醉并行腹主动脉采血致死,取出肺组织。术后观察大鼠一般情况,检测各组血清Nrf2、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、丙二醛(MDA)含量,RT-PCR法检测肺组织中Nrf2 m RNA表达量。结果中药组的血清Nrf2含量及Nrf2 mRNA表达水平高于模型组(P <0.01)。模型组大鼠血清MDA含量与正常组、假手术组及中药组相比明显升高(P <0.05),血清SOD含量及血清GSH含量与正常组、假手术组及中药组相比明显降低(P <0.05)。结论通腑平喘方对脓毒症急性肺损伤大鼠有保护作用,其机制可能与其激活Nrf2,上调SOD、GSH有关,起到抑制氧化应激反应、减轻肺组织损伤的作用。
Objectives: To observe the effect of Tongfu Pingchuan Decoction on the expression of Nrf2 in rats with sepsis-induced acute lung injury,and to explore its anti-oxidative stress. Methods: CLP was used to make the sepsis model. 40 adult male SD rats were randomly divided into 4 groups,the normal group,TCM group,sham operation group and the model group. In the sham-operated group,the cecum was removed after laparotomy and then returned to the abdominal cavity. The model group was given 0.9% sodium chloride injection immediately after CLP. Normal saline was intragastrically administered in the normal group,sham operation group and the model group 6 hours and 12 hours after CLP. The herbal medicine was intragastrically administered with Tongfu Pingchuan Decoction immediately after CLP,as well as 6 hours and 12 hours after CLP. The general condition of the rats was observed after operation. Serum Nrf2,SOD,GSH and MDA levels were detected in each group. RTPCR was used to detect the expression of Nrf2 mRNA in the lung tissue. Results: In TCM group,Nrf2 mRNA expression and serum Nrf2 level were higher than those in the model group(P < 0.01). In the model group,the serum MDA level was significantly higher than that in the normal group,the sham operation group and TCM group(P < 0.05),and the serum SOD and serum GSH levels were significantly lower(P < 0.05). Conclusions:Tongfu Pingchuan Decoction has a protective effect on rats with sepsis-induced acute lung injury,whose mechanism may be related to the activation of Nrf2 and up-regulation of SOD and GSH,which may inhibit oxidative stress and reduce lung injury.
Objectives: To observe the effect of Tongfu Pingchuan Decoction on the expression of Nrf2 in rats with sepsis-induced acute lung injury,and to explore its anti-oxidative stress. Methods: CLP was used to make the sepsis model. 40 adult male SD rats were randomly divided into 4 groups,the normal group,TCM group,sham operation group and the model group. In the sham-operated group,the cecum was removed after laparotomy and then returned to the abdominal cavity. The model group was given 0.9% sodium chloride injection immediately after CLP. Normal saline was intragastrically administered in the normal group,sham operation group and the model group 6 hours and 12 hours after CLP. The herbal medicine was intragastrically administered with Tongfu Pingchuan Decoction immediately after CLP,as well as 6 hours and 12 hours after CLP. The general condition of the rats was observed after operation. Serum Nrf2,SOD,GSH and MDA levels were detected in each group. RTPCR was used to detect the expression of Nrf2 mRNA in the lung tissue. Results: In TCM group,Nrf2 mRNA expression and serum Nrf2 level were higher than those in the model group(P < 0.01). In the model group,the serum MDA level was significantly higher than that in the normal group,the sham operation group and TCM group(P < 0.05),and the serum SOD and serum GSH levels were significantly lower(P < 0.05). Conclusions:Tongfu Pingchuan Decoction has a protective effect on rats with sepsis-induced acute lung injury,whose mechanism may be related to the activation of Nrf2 and up-regulation of SOD and GSH,which may inhibit oxidative stress and reduce lung injury.
引文
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[18]袁鼎山,李爱林,包玉华.还原型谷胱甘肽对脓毒症大鼠急性肺损伤的干预作用[J].中华危重症医学杂志,2013,6(6):353-358.
[2]Esper AM,Martin GS.Extending international sepsis epidemiology:the impact of organ dysfunction[J].Critical Care,2009,13(1):120-122.
[3]Huang Y,Pettitt SJ,Guo G,et al.Isolation of homozygous mutantmouse embryonic stem cells using a dual selection system[J].Nucleic Acids Res,2012,40(3):e21.
[4]庄良明.桑白皮汤治疗脓毒症呼吸衰竭的临床疗效观察[J].中外医学研究,2015,13(29):46-48.
[5]何淼,熊旭东,沈晓红.炎调方对脓毒症急性呼吸窘迫综合征(热炽营血证)患者血细胞因子的影响[J].中国中医急症,2013,22(7):1097-1099.
[6]Sweatt AJ,Levitt JE.Evolving epidemiology and definitions of the acute respiratory distress syndrome and early acute lung injury[J].Clin Chest Med,2014(35):609-624.
[7]Tsushima K,King LS,Aggarwal NR,et al.Acute lung injury review[J].Intern Med,2009(48):621-630.
[8]Singh G,Gladdy G,Chandy TT,et al.Incidence and outcome of acute lung injury and acute respiratory distress syndrome in the surgical intensive care unit[J].Indian J Crit Care Med,2014(18):659-665.
[9]Ward PA.Oxidative stress:acute and progressive lung injury[J].Ann N Y Acad Sci,2010(1203):53-59.
[10]Shah NR,Iqbal MB,Barlow A,et al.Severe physical exertion,oxidative stress,and acute lung injury[J].Clin J Sport Med2011(21):537-538.
[11]Shin MH,Moon YJ,Seo JE,et al.Reactive oxygen species produced by NADPH oxidase xanthine oxidase,and mitochondrial electron transport system mediate heat shockinduced MMP-1 and MMP-9 expression[J].Free RadicBio Med,2008,44(4):635-645.
[12]Abreu CC,Cardozo L,Mafra D.Could physical exercises modulate Nrf2-Keap1 pathway in chronic kidney disease[J]Med Hypotheses,2015(84):44-46.
[13]Devasagayam TP,Tilak JC,Boloor KK,et al.Free radicals and antioxidants in human health:current status and future prospects[J].J Assoc Physicians India,2004(52):794-804.
[14]Ozben T.Oxidative stress and apoptosis:impact on cancer therapy[J].J Pharm Sci,2007(96):2181-2196.
[15]Del RioD,Stewart AJ,Pellegrini N.A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress[J].Nutrition,Metabolism and Cardiovascular Diseases,2005,15(4):316-328.
[16]Ji L,Liu R,Zhang XD,et al.N-acetylcysteine attenuates phosgene-496 induced acute lung injury via up-regulation of Nrf2 expression[J].Inhal Toxicol,2010,22(7):535-542.
[17]赵京禹,费翔,岳琦,等.蛇床子素对脓毒症急性肺损伤大鼠的保护效应[J].中国医药导报,2013,10(28):103-106.
[18]袁鼎山,李爱林,包玉华.还原型谷胱甘肽对脓毒症大鼠急性肺损伤的干预作用[J].中华危重症医学杂志,2013,6(6):353-358.