人参皂甙RB2抑制NF-κB的激活对LPS诱导的新生小鼠急性肺损伤的免疫调节作用
|
摘要
目的:研究人参皂苷RB2对脂多糖(LPS)诱导的新生小鼠急性肺损伤的免疫功能的影响及相关分子机制。方法:将新生小鼠随机分为四组:健康组(Ctrl);人参皂苷单独处理组(GR2):健康小鼠腹腔注射GR2 50 mg/kg BW; LPS诱导组(LPS):腹腔注射0. 6 mg/kg BW的LPS,连续5 d; LPS+GR2组:模型小鼠腹腔注射人参皂苷50 mg/kg BW。HE染色观察肺组织损伤情况; TUNEL染色观察肺组织细胞凋亡; Western blot检测Caspase-3和Caspase-9的表达; ELISA检测炎症因子IL-6、IL-1β、TNF-α和IL-10的含量; Western blot检测高迁移率族蛋白B1(HMGB1)、巨噬细胞移动抑制因子(MIF)和NF-κBp 65的表达水平。结果:LPS组与对照组相比,肺组织出现明显的损伤,肺泡壁明显增厚,出现大量炎性细胞浸润;染色呈阳性的凋亡细胞比率显著提高; Caspase-3和Caspase-9的表达水平显著上调;炎症因子IL-6、IL-1β和TNF-α的含量显著上升,IL-10的含量没有明显变化; HMGB1和MIF的表达显著上调;磷酸化的NF-κB p65的表达显著上调。LPS+GR2组与LPS组相比,肺组织损伤明显得到缓解,多数肺泡结构完整,仅有少量炎性细胞浸润;凋亡细胞的比率显著下降; Caspase-3和Caspase-9的表达水平显著下调;炎症因子IL-6、IL-1β和TNF-α的含量显著下降,IL-10的含量没有明显的变化; HMGB1和MIF的表达显著下调;磷酸化的NF-κB p65的表达显著下调。结论:人参皂苷RB2抑制NF-κB的激活,进而调节LPS诱导的新生小鼠急性肺损伤的免疫反应。
Objective: To explore the effects of ginsenoside RB2 on immune function of lipopolysaccharide induced acute lung injury in neonatal mice and its molecular mechanism. Methods: Neonatal mice were randomly divided into four groups: the healthy group( Ctrl) and the ginsenoside alone treatment group( GR2) : the healthy mice were intraperitoneally injected with GR2 50 mg/kg BW; LPS induced group( LPS) : intraperitoneal injection of 0. 6 mg/kg BW LPS for 5 days; LPS+GR2 group: the model mice were intraperitoneally injected with ginsenoside 50 mg/kg BW. HE staining was used to observe the damage of lung tissue; apoptosis of lung tissue was observed by TUNEL staining; expression of Caspase-3 and Caspase-9 was detected by Western blot; ELISA was used to detect the levels of inflammation factor IL-6,IL-1β,TNF-α and IL-10. Results: Compared with the control group,the lung tissue was obviously damaged,and a large number of inflammatory cells were infiltrated. The percentage of apoptotic cells increased significantly; the expression level of Caspase-3 and Caspase-9 increased significantly; the content of inflammatory factors IL-6,IL-1β and TNF-αincreased significantly. There was no obvious change in the content of IL-10. The expression of HMGB1 and MIF was significantly upregulated,while the expression of phosphorylated NF-κB p65 was significantly upregulated. LPS + GR2 group compared with the LPS group,the lung tissue damage was significantly relieved. Most of the alveolar structures were intact,only a small amount of inflammatory cells were infiltrated,the percentage of apoptotic cells decreased significantly,the expression level of Caspase-3 and Caspase-9 decreased significantly,and the content of IL-6,IL-1β and TNF-α decreased significantly,and there was no obvious change in the content of IL-10. The expression of HMGB1 and MIF was significantly down regulated; the expression of phosphorylated NF-κB p65 was significantly downregulated. Conclusion: Ginsenoside RB2 inhibits the activation of NF-κB and further regulates the immune response of LPS induced acute lung injury in neonatal mice.
Objective: To explore the effects of ginsenoside RB2 on immune function of lipopolysaccharide induced acute lung injury in neonatal mice and its molecular mechanism. Methods: Neonatal mice were randomly divided into four groups: the healthy group( Ctrl) and the ginsenoside alone treatment group( GR2) : the healthy mice were intraperitoneally injected with GR2 50 mg/kg BW; LPS induced group( LPS) : intraperitoneal injection of 0. 6 mg/kg BW LPS for 5 days; LPS+GR2 group: the model mice were intraperitoneally injected with ginsenoside 50 mg/kg BW. HE staining was used to observe the damage of lung tissue; apoptosis of lung tissue was observed by TUNEL staining; expression of Caspase-3 and Caspase-9 was detected by Western blot; ELISA was used to detect the levels of inflammation factor IL-6,IL-1β,TNF-α and IL-10. Results: Compared with the control group,the lung tissue was obviously damaged,and a large number of inflammatory cells were infiltrated. The percentage of apoptotic cells increased significantly; the expression level of Caspase-3 and Caspase-9 increased significantly; the content of inflammatory factors IL-6,IL-1β and TNF-αincreased significantly. There was no obvious change in the content of IL-10. The expression of HMGB1 and MIF was significantly upregulated,while the expression of phosphorylated NF-κB p65 was significantly upregulated. LPS + GR2 group compared with the LPS group,the lung tissue damage was significantly relieved. Most of the alveolar structures were intact,only a small amount of inflammatory cells were infiltrated,the percentage of apoptotic cells decreased significantly,the expression level of Caspase-3 and Caspase-9 decreased significantly,and the content of IL-6,IL-1β and TNF-α decreased significantly,and there was no obvious change in the content of IL-10. The expression of HMGB1 and MIF was significantly down regulated; the expression of phosphorylated NF-κB p65 was significantly downregulated. Conclusion: Ginsenoside RB2 inhibits the activation of NF-κB and further regulates the immune response of LPS induced acute lung injury in neonatal mice.
引文
[1] Parekh D,Dancer RC,Thickett DR. Acute lung injury[J].Clinical Medicine,2011,11(6):615-618.
[2] Rubenfeld GD,Herridge MS. Epidemiology and outcomes of acute lung injury[J]. Chest,2007,131(2):554-562.
[3] Zimmerman JJ,Akhtar SR,Caldwell E,et al. Incidence and outcomes of pediatric acute lung injury[J]. Pediatrics,2009,124(1):87-95.
[4] National Heart L,Wiedemann HP,Wheeler AP,et al. Comparison of two fluid-management strategies in acute lung injury[J]. J Vascular Surg,2006,44(4):2564-2575.
[5] Mochizuki M,Yoo YC,Matsuzawa K,et al. Inhibitory effect of tumor metastasis in mice by saponins,ginsenoside-rb2,20(r)-and20(s)-ginsenoside-rg3,of red ginseng[J]. Biol Pharm Bull,1995,18(9):1197-1202.
[6] Lee KT,Jung TW,Lee HJ,et al. The antidiabetic effect of ginsenoside rb2 via activation of ampk[J]. Arch Pharmacal Res,2011,34(7):1201.
[7] Huang Q,Wang T,Yang L,et al. Ginsenoside rb2 alleviates hepatic lipid accumulation by restoring autophagy via induction of sirt1 and activation of ampk[J]. Int J Mol Sci,2017,18(5):1063.
[8] Butt Y,Kurdowska A,Allen TC. Acute lung injury:A clinical and molecular review[J]. Arch Pathol Lab Med,2016,140(4):345-350.
[9] Afshari A,Brok J,Mller AM,et al. Aerosolized prostacyclin for acute lung injury(ali)and acute respiratory distress syndrome(ards)[J]. Cochrane Database Syst Rev,2017,65(8):CD007733.
[10] Kang HS,Park HM,Go HK,et al. Effect of antioxidant in an acute lung injury animal model[J]. Korean J Chem Engineer,2012,29(11):1591-1596.
[11] Chen Y,Zhou JH,Yang-Yao OU,et al. Animal model of acute lung injury[J]. J Traumatic Surg,2013,15(5):466-469.
[12] Wang J,Qiao L,Li S,et al. Protective effect of ginsenoside rb1against lung injury induced by intestinal ischemia-reperfusion in rats[J]. Molecules,2013,18(1):1214.
[13] Yuan Q,Jiang Y,Ma T,et al. Attenuating effect of ginsenoside rb1 on lps-induced lung injury in rats[J]. J Inflammation,2014,11(1):40.
[14] Park SH. Ginsenosides rb2 protects high glucose-induced apoptosis in arpe cells[J]. Lab Animal Res,2008,24(1):119-123.
[15] Gao B,Huang Q,Jie Q,et al. Ginsenoside-rb2 inhibits dexamethasone-induced apoptosis through promotion of gpr120induction in bone marrow-derived mesenchymal stem cells[J].Stem Cells Development,2015,24(6):781-790.
[16] Shalini S,Dorstyn L,Dawar S,et al. Old,new and emerging functions of caspases[J]. Cell Death Differentiation,2015,22(4):526-539.
[17] Jimenez Fernandez D,Lamkanfi M. Inflammatory caspases:Key regulators of inflammation and cell death[J]. Biol Chem,2015,396(3):193-203.
[18] Manicone AM. Role of the pulmonary epithelium and inflammatory signals in acute lung injury[J]. Expert Rev Clin Immunol,2014,5(1):63-75.
[19] Wu CF,Bi XL,Yang JY,et al. Differential effects of ginsenosides on no and tnf-alpha production by lps-activated n9 microglia[J]. Int Immunopharmacol,2007,7(3):313-320.
[20] Kim DH,Moon YS,Lee TH,et al. The inhibitory effect of ginseng saponins on the stress-induced plasma interleukin-6 level in mice[J]. Neurosci Lett,2003,353(1):13-16.
[21] Bhargava M,Wendt C. Biomarkers in acute lung injury[J].Translational Res,2014,159(4):205-217.
[22] Standiford TJ,Ward PA. Therapeutic targeting of acute lung injury and ards[J]. Translational Res J Lab Clin Med,2016,167(1):183.
[23] Venereau E,De LF,Mezzapelle R,et al. Hmgb1 as biomarker and drug target[J]. Pharmacological research,2016,111(534-544.
[24] Roger T,David J,Glauser MP,et al. Mif regulates innate immune responses through modulation of toll-like receptor 4[J]. Nature,2015,414(6866):920.
[25] White AF,Demchenko AV. Modulating lps signal transduction at the lps receptor complex with synthetic lipid a analogues[J].Adv Carbohydrate Chem Biochem,2014,71:339-389.
[26] Kell DB,Pretorius E. On the translocation of bacteria and their lipopolysaccharides between blood and peripheral locations in chronic,inflammatory diseases:The central roles of lps and lpsinduced cell death[J]. Integr Biol,2015,7(11):1339-1377.
[27] Plociennikowska A,Hromada-Judycka A,Borzecka K,et al. Cooperation of tlr4 and raft proteins in lps-induced pro-inflammatory signaling[J]. Cell Mol Life Sci,2015,72(3):557-581.
[28] Lu YC,Yeh WC,Ohashi PS. Lps/tlr4 signal transduction pathway[J]. Cytokine,2008,42(2):145-151.
[29] Sun SC. The noncanonical nf-kappab pathway[J]. Immunol Rev,2012,246(1):125-140.
[2] Rubenfeld GD,Herridge MS. Epidemiology and outcomes of acute lung injury[J]. Chest,2007,131(2):554-562.
[3] Zimmerman JJ,Akhtar SR,Caldwell E,et al. Incidence and outcomes of pediatric acute lung injury[J]. Pediatrics,2009,124(1):87-95.
[4] National Heart L,Wiedemann HP,Wheeler AP,et al. Comparison of two fluid-management strategies in acute lung injury[J]. J Vascular Surg,2006,44(4):2564-2575.
[5] Mochizuki M,Yoo YC,Matsuzawa K,et al. Inhibitory effect of tumor metastasis in mice by saponins,ginsenoside-rb2,20(r)-and20(s)-ginsenoside-rg3,of red ginseng[J]. Biol Pharm Bull,1995,18(9):1197-1202.
[6] Lee KT,Jung TW,Lee HJ,et al. The antidiabetic effect of ginsenoside rb2 via activation of ampk[J]. Arch Pharmacal Res,2011,34(7):1201.
[7] Huang Q,Wang T,Yang L,et al. Ginsenoside rb2 alleviates hepatic lipid accumulation by restoring autophagy via induction of sirt1 and activation of ampk[J]. Int J Mol Sci,2017,18(5):1063.
[8] Butt Y,Kurdowska A,Allen TC. Acute lung injury:A clinical and molecular review[J]. Arch Pathol Lab Med,2016,140(4):345-350.
[9] Afshari A,Brok J,Mller AM,et al. Aerosolized prostacyclin for acute lung injury(ali)and acute respiratory distress syndrome(ards)[J]. Cochrane Database Syst Rev,2017,65(8):CD007733.
[10] Kang HS,Park HM,Go HK,et al. Effect of antioxidant in an acute lung injury animal model[J]. Korean J Chem Engineer,2012,29(11):1591-1596.
[11] Chen Y,Zhou JH,Yang-Yao OU,et al. Animal model of acute lung injury[J]. J Traumatic Surg,2013,15(5):466-469.
[12] Wang J,Qiao L,Li S,et al. Protective effect of ginsenoside rb1against lung injury induced by intestinal ischemia-reperfusion in rats[J]. Molecules,2013,18(1):1214.
[13] Yuan Q,Jiang Y,Ma T,et al. Attenuating effect of ginsenoside rb1 on lps-induced lung injury in rats[J]. J Inflammation,2014,11(1):40.
[14] Park SH. Ginsenosides rb2 protects high glucose-induced apoptosis in arpe cells[J]. Lab Animal Res,2008,24(1):119-123.
[15] Gao B,Huang Q,Jie Q,et al. Ginsenoside-rb2 inhibits dexamethasone-induced apoptosis through promotion of gpr120induction in bone marrow-derived mesenchymal stem cells[J].Stem Cells Development,2015,24(6):781-790.
[16] Shalini S,Dorstyn L,Dawar S,et al. Old,new and emerging functions of caspases[J]. Cell Death Differentiation,2015,22(4):526-539.
[17] Jimenez Fernandez D,Lamkanfi M. Inflammatory caspases:Key regulators of inflammation and cell death[J]. Biol Chem,2015,396(3):193-203.
[18] Manicone AM. Role of the pulmonary epithelium and inflammatory signals in acute lung injury[J]. Expert Rev Clin Immunol,2014,5(1):63-75.
[19] Wu CF,Bi XL,Yang JY,et al. Differential effects of ginsenosides on no and tnf-alpha production by lps-activated n9 microglia[J]. Int Immunopharmacol,2007,7(3):313-320.
[20] Kim DH,Moon YS,Lee TH,et al. The inhibitory effect of ginseng saponins on the stress-induced plasma interleukin-6 level in mice[J]. Neurosci Lett,2003,353(1):13-16.
[21] Bhargava M,Wendt C. Biomarkers in acute lung injury[J].Translational Res,2014,159(4):205-217.
[22] Standiford TJ,Ward PA. Therapeutic targeting of acute lung injury and ards[J]. Translational Res J Lab Clin Med,2016,167(1):183.
[23] Venereau E,De LF,Mezzapelle R,et al. Hmgb1 as biomarker and drug target[J]. Pharmacological research,2016,111(534-544.
[24] Roger T,David J,Glauser MP,et al. Mif regulates innate immune responses through modulation of toll-like receptor 4[J]. Nature,2015,414(6866):920.
[25] White AF,Demchenko AV. Modulating lps signal transduction at the lps receptor complex with synthetic lipid a analogues[J].Adv Carbohydrate Chem Biochem,2014,71:339-389.
[26] Kell DB,Pretorius E. On the translocation of bacteria and their lipopolysaccharides between blood and peripheral locations in chronic,inflammatory diseases:The central roles of lps and lpsinduced cell death[J]. Integr Biol,2015,7(11):1339-1377.
[27] Plociennikowska A,Hromada-Judycka A,Borzecka K,et al. Cooperation of tlr4 and raft proteins in lps-induced pro-inflammatory signaling[J]. Cell Mol Life Sci,2015,72(3):557-581.
[28] Lu YC,Yeh WC,Ohashi PS. Lps/tlr4 signal transduction pathway[J]. Cytokine,2008,42(2):145-151.
[29] Sun SC. The noncanonical nf-kappab pathway[J]. Immunol Rev,2012,246(1):125-140.