人参皂甙Rb1对机械通气大鼠肺组织NF-κB和iNOs表达的影响
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摘要
目的观察人参皂甙Rb1对机械通气肺损伤(VILI)大鼠炎症信号通路蛋白NF-κB及诱导型一氧化氮合酶(iNOs)的影响。方法将30只SD大鼠随机分为假手术组(C组)、VILI组和人参皂甙Rb1预处理组(Rb1组),每组10只。后两组大鼠潮气量40ml/kg机械通气4h建立大鼠VILI模型,Rb1组在通气前30min尾静脉注射40mg/kg人参皂甙Rb1,其他两组予等容量0.9%氯化钠溶液。观察各组大鼠肺组织病理改变,肺组织湿/干重比(W/D),收集肺泡灌洗液(BALF),采用ELISA法测定BALF中TNF-α、IL-6水平,硝酸盐还原酶法检测一氧化氮(NO)水平;RT-PCR测定肺组织中iNOs mRNA的表达水平,Western blot检测肺组织的NF-κB蛋白水平。结果与C组相比,VILI组肺组织病理损伤严重,肺组织W/D显著增加,BALF中TNF-α、IL-6、NO水平增加,iNOs mRNA表达增加,NF-κB蛋白表达上调;与VILI组比较,Rb1组肺组织病理损伤程度减轻,肺组织W/D降低,BALF中TNF-α、IL-6、NO水平降低,iNOs mRNA表达降低,NF-κB蛋白表达下调。结论人参皂甙Rb1可下调机械通气大鼠NF-κB及iNOs活性,降低促炎因子TNF-α、IL-6、NO释放,减轻肺损伤。
Objective To investigate the effects of ginsenoside Rb1 on expression of NF-κB and iNOs in lung tissue of rats with ventilator-induced lung injury. Methods Thirty SD rats were randomly divided into 3 group:sham operation group(C group),ventilator-induced lung injury group(VILI group) and ginsenoside Rb1 pretreatment group(Rb1 group). The lung injury was induced by mechanical ventilation with a high tidal volume of 40 ml/kg for 4 h. The Rb1 group was given 40 mg/kg ginsenoside Rb1 by tail vein injection30 min before mechanical ventilation. The other two groups were given equal volume of normal saline. After 4 h of mechanical ventilation, the pathological changes of lung tissue and lung wet/dry weight ratio(W/D) were observed. The brochoalveolar lavage fluid(BALF) was collected, TNF-α and IL-6 in BALF were determined by ELISA; the concentration of nitric oxide(NO) was measured by nitrate reductase method. The expressions of iNOs mRNA and NF-κB protein in lung tissue were determined by RT-PCR and Western blot,respectively. Results Compared with group C, the pathological damage of lung tissue in VILI group was severe, the W/D of lung was increased significantly, the concentrations of TNF-α, IL-6 and NO in BALF were increased, the expression of iNOs mRNA and NF-κB protein in lung tissue was increased. Compared with VILI group, the pathological damage of lung tissue was attenuated in Rb1 group, the W/D of lung was decreased, the concentrations of TNF-α, IL-6 and NO in BALF were decreased, the expression of iNOs mRNA and NF-κB protein in lung tissue was also decreased. Conclusion Ginsenoside Rb1 can down-regulate the expression of NF-κB and iNOs in the inflammatory pathway, decrease the concentrations of pro-inflammatory factors TNF-α, IL-6 and NO, and attenuate the lung injury in rats with ventilatorinduced lung injury.
Objective To investigate the effects of ginsenoside Rb1 on expression of NF-κB and iNOs in lung tissue of rats with ventilator-induced lung injury. Methods Thirty SD rats were randomly divided into 3 group:sham operation group(C group),ventilator-induced lung injury group(VILI group) and ginsenoside Rb1 pretreatment group(Rb1 group). The lung injury was induced by mechanical ventilation with a high tidal volume of 40 ml/kg for 4 h. The Rb1 group was given 40 mg/kg ginsenoside Rb1 by tail vein injection30 min before mechanical ventilation. The other two groups were given equal volume of normal saline. After 4 h of mechanical ventilation, the pathological changes of lung tissue and lung wet/dry weight ratio(W/D) were observed. The brochoalveolar lavage fluid(BALF) was collected, TNF-α and IL-6 in BALF were determined by ELISA; the concentration of nitric oxide(NO) was measured by nitrate reductase method. The expressions of iNOs mRNA and NF-κB protein in lung tissue were determined by RT-PCR and Western blot,respectively. Results Compared with group C, the pathological damage of lung tissue in VILI group was severe, the W/D of lung was increased significantly, the concentrations of TNF-α, IL-6 and NO in BALF were increased, the expression of iNOs mRNA and NF-κB protein in lung tissue was increased. Compared with VILI group, the pathological damage of lung tissue was attenuated in Rb1 group, the W/D of lung was decreased, the concentrations of TNF-α, IL-6 and NO in BALF were decreased, the expression of iNOs mRNA and NF-κB protein in lung tissue was also decreased. Conclusion Ginsenoside Rb1 can down-regulate the expression of NF-κB and iNOs in the inflammatory pathway, decrease the concentrations of pro-inflammatory factors TNF-α, IL-6 and NO, and attenuate the lung injury in rats with ventilatorinduced lung injury.
引文
[1]Kneyber MC,Zhang H,Slutsky AS.Ventilator-induced lung injury.Similarity and differences between children and adults[J].Am JRespir Crit Care Med,2014,190(3):258-265.DOI:10.1164/rccm.201401-0168CP.
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[8]Shu Yu-Sheng,Tao Wei,Miao Qian-Bing,et al.Improvement of ventilation-induced lung injury in a rodent model by inhibition of inhibitoryκB kinase[J].J Trauma Acute Care Surg,2014,76(6):1417-1424.DOI:10.1097/TA.0000000000000229.
[9]Held H D,Boettcher S,Hamann L,et al.Ventilation-induced chemokine and cytokine release is associated with activation of nuclear factor-kappaB and is blocked by steroids[J].Am J Respir Crit Care Med,2001,163(3 Pt 1):711-716.DOI:10.1164/ajrccm.163.3.2003001.
[10]Wang Jin,Qiao Lifen,Li Yongsheng,et al.Ginsenoside Rb1attenuates intestinal ischemia-reperfusion-induced liver injury by inhibiting NF-kappaB activation[J].Exp Mol Med,2008,40(6):686-698.DOI:10.3858/emm.2008.40.6.686.
[11]Zhou Ping,Lu Shan,Luo Yun,et al.Attenuation of TNF-αInduced Inflammatory Injury in Endothelial Cells by Ginsenoside Rb1 via Inhibiting NF-κB,JNK and p38 Signaling Pathways[J].Front Pharmacol,2017,8:464.DOI:10.3389/fphar.2017.00464.
[12]Peng Xinqi,Abdulnour Raja-Elie E,Sammani Saad,et al.Inducible nitric oxide synthase contributes to ventilator-induced lung injury[J].Am J Respir Crit Care Med,2005,172(4):470-479.DOI:10.1164/rccm.200411-1547OC.
[13]Hatano E,Bennett B L,Manning AM,et al.NF-kappaBstimulates inducible nitric oxide synthase to protect mouse hepatocytes from TNF-alpha-and Fas-mediated apoptosis[J].Gastroenterology,2001,120:1251-1262.DOI:10.1053/gast.2001.23239.
[2]Ngiam N,Kavanagh BP.Ventilator-induced lung injury:the role of gene activation[J].Curr Opin Crit Care,2012,18(1):16-22.DOI:10.1097/MCC.0b013e32834e7d00.
[3]Park EK,Shin YW,Lee HU,et al.Inhibitory effect of ginsenoside Rb1 and compound K on NO and prostaglandin E2 biosyntheses of RAW264.7 cells induced by lipopolysaccharide[J].Biol Pharm Bull,2005,28(4):652-656.DOI:10.1248/bpb.28.652.
[4]单鸳露,金立达,王良荣,等.环氧合酶-2抑制剂对大鼠机械通气所致肺损伤细胞因子及氧化应激的影响[J].实用医学杂志,2013,29(16):2616-2618.
[5]Rocco PR,Dos SC,Pelosi P.Pathophysiology of ventilator-associated lung injury[J].Curr Opin Anaesthesiol,2012,25(2):123-130.DOI:10.1097/ACO.0b013e32834f8c7f.
[6]Vallabhapurapu Sivakumar,Karin Michael.Regulation and function of NF-kappa B transcription factors in the immune system[J].Annu Rev Immunol,2009,27:693-733.DOI:10.1146/annurev.immunol.021908.132641.
[7]Dos SCC,Slutsky AS.Invited review:mechanisms of ventilatorinduced lung injury:a perspective[J].J Appl Physiol(1985),2000,89(4):1645-1655.DOI:10.1152/jappl.2000.89.4.1645.
[8]Shu Yu-Sheng,Tao Wei,Miao Qian-Bing,et al.Improvement of ventilation-induced lung injury in a rodent model by inhibition of inhibitoryκB kinase[J].J Trauma Acute Care Surg,2014,76(6):1417-1424.DOI:10.1097/TA.0000000000000229.
[9]Held H D,Boettcher S,Hamann L,et al.Ventilation-induced chemokine and cytokine release is associated with activation of nuclear factor-kappaB and is blocked by steroids[J].Am J Respir Crit Care Med,2001,163(3 Pt 1):711-716.DOI:10.1164/ajrccm.163.3.2003001.
[10]Wang Jin,Qiao Lifen,Li Yongsheng,et al.Ginsenoside Rb1attenuates intestinal ischemia-reperfusion-induced liver injury by inhibiting NF-kappaB activation[J].Exp Mol Med,2008,40(6):686-698.DOI:10.3858/emm.2008.40.6.686.
[11]Zhou Ping,Lu Shan,Luo Yun,et al.Attenuation of TNF-αInduced Inflammatory Injury in Endothelial Cells by Ginsenoside Rb1 via Inhibiting NF-κB,JNK and p38 Signaling Pathways[J].Front Pharmacol,2017,8:464.DOI:10.3389/fphar.2017.00464.
[12]Peng Xinqi,Abdulnour Raja-Elie E,Sammani Saad,et al.Inducible nitric oxide synthase contributes to ventilator-induced lung injury[J].Am J Respir Crit Care Med,2005,172(4):470-479.DOI:10.1164/rccm.200411-1547OC.
[13]Hatano E,Bennett B L,Manning AM,et al.NF-kappaBstimulates inducible nitric oxide synthase to protect mouse hepatocytes from TNF-alpha-and Fas-mediated apoptosis[J].Gastroenterology,2001,120:1251-1262.DOI:10.1053/gast.2001.23239.