水飞蓟素通过调控TLR/NF-κB信号通路改善急性肺损伤大鼠肺功能作用研究
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摘要
目的观察水飞蓟素对急性肺损伤(ALI)大鼠肺功能的保护作用并探讨其机制。方法采用脂多糖制备ALI模型大鼠,随机分为ALI组、地塞米松(0.9 mg/kg)组、低(15 mg/kg)、中(30 mg/kg)、高(60 mg/kg)剂量水飞蓟素组、TLR4抑制剂TAK-242(0.5 mg/kg)组,对照组(等体积生理盐水)及ALI组(等体积生理盐水)。灌胃给药24 h后,分析各组大鼠动脉血气指标;苏木素-伊红(HE)染色观察肺组织形态;酶联免疫吸附(ELISA)法检测肺泡灌洗液中单核细胞趋化因子(MCP-1)、白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平;蛋白印迹(WB)法检测肺组织Toll样受体4(TLR4)、核转录因子-κB(NF-κB)表达情况。结果与对照组比较,ALI组大鼠W/D、Pa CO_2水平增高、Pa O_2水平明显下降(P <0.05);肺组织现细胞破碎等病变;肺泡灌洗液中SOD、MPO活性降低(P <0.05),MCP-1、IL-1β、TNF-α水平增高(P <0.05);肺组织TLR4表达上调、NF-κB核移位增加(P <0.05)。与ALI组比较,低、中、高剂量水飞蓟素组和TAK-242组大鼠大鼠W/D、PaCO_2水平降低、Pa O_2水平明显增高(P <0.05),肺组织病变减轻,肺泡灌洗液中SOD、MPO活性升高(P <0.05),MCP-1、IL-1β、TNF-α水平降低;肺组织TLR4表达下调、NF-κB核移位减少(P <0.05)。结论水飞蓟素可能通过调控TLR4/NF-κB通路抑制氧化应激反应和炎症反应,保护急性肺损伤大鼠肺功能。
Objective: To observe the protective effect of Silymarin on lung function in rats with acute lung injury(ALI) and explore its mechanism. Methods: ALI model rats were prepared by lipopolysaccharide,and randomly divided into ALI group,Dexamethasone(0. 9 mg/kg),low(15 mg/kg),middle(30 mg/kg),high(60 mg/kg)dose of Silymarin groups and TLR4 inhibitor TAK-242(0.5 mg/kg) group. The blank control group and ALI group were given gastric lavage with equal volume of normal saline. After administration of 24 h,the arterial blood gas in each group was analyzed;the lung tissue morphology was observed by HE staining;the levels of MCP-1,IL-1βand TNF-α in lung lavage fluid were detected by ELISA;TLR4 andNF-κB were detected by WB. Results: Compared with the control group,the W/D,Pa CO_2 levels were increased,and the level of PaO_2 was decreased significantly in ALI group(P < 0.05);cell breakage and other pathological changes appeared in lung tissue;the activities of SOD and MPO in lung lavage fluid decreased(P < 0.05),and the levels of MCP-1,IL-1β and TNF-α increased(P < 0.05);the expression of TLR4 in lung tissue was up-regulated and NF-κB nuclear shift increased(P < 0.05).Compared with ALI group,the W/D,Pa CO_2 levels in low,middle and high doses of Silymarin groups and TAK-242 group were decreased,and the level of PaO_2 was increased significantly(P < 0.05),lung lesions were alleviated,the SOD and MPO activities in lung lavage fluid were increased(P < 0.05),MCP-1,IL-1β and TNF-α levels were decreased;the expression of TLR4 in lung tissue was down-regulated and NF-κB nuclear shift decreased(P< 0.05). Conclusion: Silymarin may inhibit oxidative stress and inflammation by regulating the TLR4/NF-κB pathway,and then protect the lung function of rats with acute lung injury.
Objective: To observe the protective effect of Silymarin on lung function in rats with acute lung injury(ALI) and explore its mechanism. Methods: ALI model rats were prepared by lipopolysaccharide,and randomly divided into ALI group,Dexamethasone(0. 9 mg/kg),low(15 mg/kg),middle(30 mg/kg),high(60 mg/kg)dose of Silymarin groups and TLR4 inhibitor TAK-242(0.5 mg/kg) group. The blank control group and ALI group were given gastric lavage with equal volume of normal saline. After administration of 24 h,the arterial blood gas in each group was analyzed;the lung tissue morphology was observed by HE staining;the levels of MCP-1,IL-1βand TNF-α in lung lavage fluid were detected by ELISA;TLR4 andNF-κB were detected by WB. Results: Compared with the control group,the W/D,Pa CO_2 levels were increased,and the level of PaO_2 was decreased significantly in ALI group(P < 0.05);cell breakage and other pathological changes appeared in lung tissue;the activities of SOD and MPO in lung lavage fluid decreased(P < 0.05),and the levels of MCP-1,IL-1β and TNF-α increased(P < 0.05);the expression of TLR4 in lung tissue was up-regulated and NF-κB nuclear shift increased(P < 0.05).Compared with ALI group,the W/D,Pa CO_2 levels in low,middle and high doses of Silymarin groups and TAK-242 group were decreased,and the level of PaO_2 was increased significantly(P < 0.05),lung lesions were alleviated,the SOD and MPO activities in lung lavage fluid were increased(P < 0.05),MCP-1,IL-1β and TNF-α levels were decreased;the expression of TLR4 in lung tissue was down-regulated and NF-κB nuclear shift decreased(P< 0.05). Conclusion: Silymarin may inhibit oxidative stress and inflammation by regulating the TLR4/NF-κB pathway,and then protect the lung function of rats with acute lung injury.
引文
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[15]Cui L,Feng L,Zhang ZH,et al.The anti-inflammation effect of baicalin on experimental colitis through inhibiting TLR4/NF-κB pathway activation[J].Int Immunopharmacol,2014,23(1):294-303.
[16]Di GM,Zanoni I.Toll-like receptor co-receptors as master regulators of the immune response[J].Mol Immunol,2015,63(2):143-152.
[17]Li Q,Jiang Y,Jiang S,et al.Sesamin attenuates lipopolysaccharide-induced acute lung injury by inhibition of TLR4 signaling pathways[J].Inflammation,2016,39(1):1-6.
[18]Zhang Y,Lu Y,Ma L,et al.Activation of vascular endothelial growth factor receptor-3 in macrophages restrains TLR4-NF-κB signaling and protects against endotoxin shock[J].Immunity,2014,40(4):501-514.
[19]Chi X,Wei X,Gao W,et al.Dexmedetomidine ameliorates acute lung injury following orthotopic autologous liver transplantation in rats probably by inhibiting Toll-like receptor 4-nuclear factor kappa B signaling[J].J Transl Med,2015,13(1):190-197.
[20]Jiang Q,Yi M,Guo Q,et al.Protective effects of polydatin on lipopolysaccharide-induced acute lung injury through TLR4-MyD88-NF-κB pathway[J].Int Immunopharmacol,2015,29(2):370-376.
[2]Mario M,Jan Adriaan G,Henrik S,et al.Characterization of inflammation in a rat model of acute lung injury after repeated pulmonary lavage[J].Exp Lung Res,2015,41(8):466-476.
[3]章梦丽,李文强.二甲双胍对脓毒症急性肺损伤大鼠肺组织线粒体氧化应激的影响[J].中国急救医学,2017,37(11):973-976.
[4]Theodosiou E,PurchartováK,Stamatis H,et al.Bioavailability of silymarin flavonolignans:drug formulations and biotransformation[J].Phytochem Rev,2014,13(1):1-18.
[5]Surai PF.Silymarin as a natural antioxidant:an overview of the current evidence and perspectives[J].Antioxidants,2015,4(1):204-247.
[6]孙谋,孙同文,余言午,等.脂肪干细胞对脂多糖诱导急性肺损伤大鼠炎性因子表达的影响[J].中华危重病急救医学,2016,28(10):911-915.
[7]胡晓旻,刘刚,刘超,等.间充质干细胞缓解大鼠急性肺损伤的分子机制[J].中国老年学杂志,2017,37(15):3664-3667.
[8]张珂,张琬啉,李洪图.水飞蓟素对大鼠肺缺血再灌注损伤的保护作用[J].中华实验外科杂志,2016,33(8):1935-1937.
[9]Farjad E,Momeni HR.Silymarin Ameliorates oxidative stress and enhances antioxidant defense system capacity in cadmium-treated mice[J].Cell J,2018,20(3):422-426.
[10]孙红,赵自刚.活性氧在失血性休克引起急性肺损伤中的作用机制[J].中国急救医学,2017,37(1):92-96.
[11]Aksu B,Ayvaz S,Aksu F,et al.Effects of sphingosylphosphorylcholine against oxidative stress and acute lung injury induced by pulmonary contusion in rats[J].J Pediatr Surg,2015,50(4):591-597.
[12]Kumari A,Tyagi N,Dash D,et al.Intranasal curcumin ameliorates lipopolysaccharide-induced acute lung injury in mice[J].Inflammation,2015,38(3):1103-1112.
[13]Ronchi CF,Ferreira AL,Campos FJ,et al.Interactive effects of mechanical ventilation,inhaled nitric oxide and oxidative stress in acute lung injury[J].Respir Physiol Neurobiol,2014,190(1):118-123.
[14]Gerin F,Sener U,Erman H,et al.The effects of quercetin on acute lung injury and biomarkers of inflammation and oxidative stress in the rat model of sepsis[J].Inflammation,2016,39(2):700-705.
[15]Cui L,Feng L,Zhang ZH,et al.The anti-inflammation effect of baicalin on experimental colitis through inhibiting TLR4/NF-κB pathway activation[J].Int Immunopharmacol,2014,23(1):294-303.
[16]Di GM,Zanoni I.Toll-like receptor co-receptors as master regulators of the immune response[J].Mol Immunol,2015,63(2):143-152.
[17]Li Q,Jiang Y,Jiang S,et al.Sesamin attenuates lipopolysaccharide-induced acute lung injury by inhibition of TLR4 signaling pathways[J].Inflammation,2016,39(1):1-6.
[18]Zhang Y,Lu Y,Ma L,et al.Activation of vascular endothelial growth factor receptor-3 in macrophages restrains TLR4-NF-κB signaling and protects against endotoxin shock[J].Immunity,2014,40(4):501-514.
[19]Chi X,Wei X,Gao W,et al.Dexmedetomidine ameliorates acute lung injury following orthotopic autologous liver transplantation in rats probably by inhibiting Toll-like receptor 4-nuclear factor kappa B signaling[J].J Transl Med,2015,13(1):190-197.
[20]Jiang Q,Yi M,Guo Q,et al.Protective effects of polydatin on lipopolysaccharide-induced acute lung injury through TLR4-MyD88-NF-κB pathway[J].Int Immunopharmacol,2015,29(2):370-376.