蛇床子素对失血性休克所致肺损伤大鼠的保护作用及机制
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摘要
目的探讨蛇床子素(Osthole)对失血性休克所导致的肺损伤大鼠的保护作用及其治疗机制。方法将SPF级SD大鼠50只,体重200-250g,随机分为假手术组(Sham组)、失血性休克组(HS组)、蛇床子素组(Ost组)、NF-κB抑制剂组(PDTC组)、蛇床子素联合NF-κB抑制剂组(Ost+PDTC组),每组10只。利用股动脉放血法制作大鼠失血性休克模型,自体血回输复苏;Sham组不放血,Ost组于建模后腹腔注射蛇床子素25mg/kg,于HS发生后4h处死大鼠。HE染色观察大鼠肺组织病理变化;检测大鼠各组肺湿/干重比;检测肺组织SOD和MDA水平;ELISA法检测大鼠血清中IL-1β, IL-6, TNF-α;Western Blot法检测各组大鼠肺组织中NF-κB p65, IκB-α, p-IκB-α蛋白表达水平。结果 HS组大鼠肺损伤严重,肺组织中SOD水平降低、MDA水平升高(P<0.05),血清中IL-1β、IL-6及TNF-α水平升高(P<0.05),蛇床子素可以抑制失血性休克引发的肺损伤;Ost组大鼠肺湿/干重比显著降低;肺组织SOD水平升高、MDA水平降低(P<0.05);血清中IL-1β、IL-6和TNF-α水平均降低(P<0.05);同时, NF-κBp65、 p-IκB-α蛋白表达降低, IκB-α蛋白表达升高(P<0.05),且加入NF-κB抑制剂后,蛇床子素对炎症因子的改善作用受到抑制,对NF-κB信号通路相关蛋白的调控也受到抑制。结论蛇床子素有效减轻失血性休克导致的急性肺损伤,与抑制NF-κB信号通路介导的炎症反应相关。
Objective To explore the effect of osthole on acute lung injury and its possible mechanism in hemorrhagic shock rat model. Methods Fifty SPF grade SD rats, weighing 200-250 g, were randomly divided into shamoperated group(Sham), hemorrhagic shock group(HS group), osthole-treated group(Ost group), NF-κB inhibitor group(PDTC group) and osthole+inhibitor group(Ost+PDTC) group, 10 rats in each group. The HS model of rats was established by femoral arterial transfusion. The autologous blood was resuscitated in the HS group, Ost group was intraperitoneally injected with Ost(25 mg/kg) and the rats were sacrificed 4 hours after the HS. HE staining was used to observe the pathological changes of lung tissue; the ratio of lung wet/dry weight was detected; and the levels of SOD and MDA in lung tissue was tested. The contents of IL-1β and IL-6, TNF-α in serum were detected by ELISA. Western Blot was used to detect the expression of NF-κB p65, IκB-α and p-IκB-α proteins. Results In the HS group, the lung injury was severe, the level of SOD decreased, the level of MDA increased in the lung tissue, and the levels of IL-1β, IL-6 and TNF-α in the serum increased, Ost inhibited lung injury caused by hemorrhagic shock. The lung wet/dry weight ratio of Ost group decreased significantly, the level of SOD increased, but the level of MDA decreased in lung tissue, IL-1β, IL-6 and TNF-α levels in serum were lower; At the same time, NF-κB p65, p-IκB-α protein expression decreased, IκB-α protein expression increased(P<0.05), and NF-κB inhibitor inhibited the effect of Ost on the inflammatory factor, and the regulation of the NF-κB signaling pathway-related protein was also inhibited. Conclusion Osthole effectively alleviates the acute lung injury caused by hemorrhagic shock, which is related to the inhibition of inflammatory response mediated by NF-κB signaling pathway.
Objective To explore the effect of osthole on acute lung injury and its possible mechanism in hemorrhagic shock rat model. Methods Fifty SPF grade SD rats, weighing 200-250 g, were randomly divided into shamoperated group(Sham), hemorrhagic shock group(HS group), osthole-treated group(Ost group), NF-κB inhibitor group(PDTC group) and osthole+inhibitor group(Ost+PDTC) group, 10 rats in each group. The HS model of rats was established by femoral arterial transfusion. The autologous blood was resuscitated in the HS group, Ost group was intraperitoneally injected with Ost(25 mg/kg) and the rats were sacrificed 4 hours after the HS. HE staining was used to observe the pathological changes of lung tissue; the ratio of lung wet/dry weight was detected; and the levels of SOD and MDA in lung tissue was tested. The contents of IL-1β and IL-6, TNF-α in serum were detected by ELISA. Western Blot was used to detect the expression of NF-κB p65, IκB-α and p-IκB-α proteins. Results In the HS group, the lung injury was severe, the level of SOD decreased, the level of MDA increased in the lung tissue, and the levels of IL-1β, IL-6 and TNF-α in the serum increased, Ost inhibited lung injury caused by hemorrhagic shock. The lung wet/dry weight ratio of Ost group decreased significantly, the level of SOD increased, but the level of MDA decreased in lung tissue, IL-1β, IL-6 and TNF-α levels in serum were lower; At the same time, NF-κB p65, p-IκB-α protein expression decreased, IκB-α protein expression increased(P<0.05), and NF-κB inhibitor inhibited the effect of Ost on the inflammatory factor, and the regulation of the NF-κB signaling pathway-related protein was also inhibited. Conclusion Osthole effectively alleviates the acute lung injury caused by hemorrhagic shock, which is related to the inhibition of inflammatory response mediated by NF-κB signaling pathway.
引文
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[11]Chao X,Zhou J,Chen T,et al.Neuroprotective effect of osthole against acute ischemic stroke on middle cerebral ischemia occlusion in rats[J].Brain Res,2010,1363(3):206-211.
[12]McCord JM,Edeas MA.SOD,oxidative stress and human pathologies:a brief history and a future vision[J].Biomed Pharmacother,2005,59(4):139-142.
[13]Sonnier DI,Makley AT,Friend LA,et al.Hemorrhagic shock induces a proinflammatory milieu in the gut lumen[J].J Surg Res,2011,170(2):272-279.
[14]Varelias A,Gartlan KH,Kreijveld E,et al.Lung parenchymaderived IL-6 promotes IL-17A-dependent acute lung injury after allogeneic stem cell transplantation[J].Blood,2015,125(15):2435-2444.
[15]Li Y,Wang LM,Xu JZ,et al.Gastrodia elata attenuates inflammatory response by inhibiting the NF-κB pathway in rheumatoid arthritis fibroblast-like synoviocytes[J].Biomed Pharmacother,2017,85(5):177-181.
[16]Zhao Y,Lee JH,Chen D,et al.DL-3-n-butylphthalide induced neuroprotection,regenerative repair,functional recovery and psychological benefits following traumatic brain injury in mice[J].Neurochem Int,2017,111(9):82-92.
[2]廖志林.急性肺损伤与失血性休克相关性研究进展[J].实用临床医学,2013,14(08):136-139.
[3]Kosaka J,Morimatsu H,Takahashi T,et al.Effects of biliverdin administration on acute lung injury induced by hemorrhagic shock and resuscitation in rats[J].PLoS One,2013,8(5):e63606.
[4]张炯,王佳,王芳,等.蛇床子素对大鼠肾脏缺血再灌注损伤的保护作用研究[J].安徽医科大学学报,2018(11):1731-1735.
[5]陶震宇,孔亮,姚璎珈,等.蛇床子素对机械性脑损伤小鼠的治疗作用研究[J].中国新药杂志,2016,25(05):589-593.
[6]Niu Y,Wan C,Zhou B,et al.Aerobic exercise relieved vascular cognitive impairment via NF-κB/miR-503/BDNF pathway[J].Am J Transl Res,2018,10(3):753-761.
[7]Jeong KY,Suh GJ,Kwon WY,et al.The therapeutic effect and mechanism of niacin on acute lung injury in a rat model of hemorrhagic shock:Down-regulation of the reactive oxygen speciesdependent nuclear factorκB pathway[J].J Trauma Acute Care Surg,2015,79(2):247-255.
[8]Santiago VR,Rzezinski AF,Nardelli LM,et al.Recruitment maneuver in experimental acute lung injury:the role of alveolar collapse and edema[J].Crit Care Med,2010,38(11):2207-2214.
[9]Miller AC,Elamin EM,Suffredini AF.Inhaled anticoagulation regimens for the treatment of smoke inhalation-associated acute lung injury:a systematic review[J].Crit Care Med,2014,42(2):413-419.
[10]Hariprashad A,Rizzolo D.Acute respiratory distress syndrome:an overview for physician assistants[J].JAAPA,2013,26(9):23-28.
[11]Chao X,Zhou J,Chen T,et al.Neuroprotective effect of osthole against acute ischemic stroke on middle cerebral ischemia occlusion in rats[J].Brain Res,2010,1363(3):206-211.
[12]McCord JM,Edeas MA.SOD,oxidative stress and human pathologies:a brief history and a future vision[J].Biomed Pharmacother,2005,59(4):139-142.
[13]Sonnier DI,Makley AT,Friend LA,et al.Hemorrhagic shock induces a proinflammatory milieu in the gut lumen[J].J Surg Res,2011,170(2):272-279.
[14]Varelias A,Gartlan KH,Kreijveld E,et al.Lung parenchymaderived IL-6 promotes IL-17A-dependent acute lung injury after allogeneic stem cell transplantation[J].Blood,2015,125(15):2435-2444.
[15]Li Y,Wang LM,Xu JZ,et al.Gastrodia elata attenuates inflammatory response by inhibiting the NF-κB pathway in rheumatoid arthritis fibroblast-like synoviocytes[J].Biomed Pharmacother,2017,85(5):177-181.
[16]Zhao Y,Lee JH,Chen D,et al.DL-3-n-butylphthalide induced neuroprotection,regenerative repair,functional recovery and psychological benefits following traumatic brain injury in mice[J].Neurochem Int,2017,111(9):82-92.