槲皮素通过抑制TLR4/NF-κB通路缓解脂多糖诱导的急性肾损伤
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摘要
目的分析槲皮素对脂多糖诱导小鼠急性肾损伤的保护作用,并初步探讨其机制。方法将40只BALB/c小鼠随机分为4组:对照组(无脂多糖及槲皮素),脂多糖组(15 mg/kg脂多糖),低剂量组(25 mg/kg槲皮素+15 mg/kg脂多糖),高剂量组(50 mg/kg槲皮素+15 mg/kg脂多糖)。槲皮素予以胃灌注预处理,1次/d,连续3 d,对照组同时予以相同体积的生理盐水。最后1次灌胃后1 h予以腹腔注射15 mg/kg的脂多糖,脂多糖干预24 h后结束实验,处死小鼠。观察小鼠肾脏组织病理变化和血肌酐、尿素氮评估肾脏损伤情况,ELISA法检测血清中TNF-α、IL-1β、IL-6的表达情况,Western blot检测肾脏胞浆中TLR-4、MyD88、TRAF-6以及核内NF-κB p65蛋白的表达。结果槲皮素可以降低脂多糖诱导的急性肾损伤的肌酐、尿素氮水平以及肾脏损伤的评分(P<0.05),降低血清中TNF-α、IL-1β、IL-6的表达(P<0.05),槲皮素抑制肾脏组织中TLR4、MyD88、TRAF-6及核内NF-κB p65蛋白的表达(P<0.05)。结论槲皮素预处理可能通过抑制TLR4/NF-κB信号通路保护脂多糖诱导的小鼠急性肾损伤。
Objective To investigate the protective effect of quercetin against lipopolysaccharide(LPS)-induced acute kidney injury(AKI) in mice and explore its mechanism. Methods Forty male BALB/c mice were randomly divided into control group(with saline treatment), 15 mg/kg LPS group, and quercetin-treated groups with intragastric quercetin treatment(once daily for 3 consecutive days) at low(25 mg/kg) and high(50 mg/kg) dose prior to 15 mg/kg LPS injection. LPS was administered by intraperitoneally injection 1 after the last gavage of quercetin. The mice were sacrificed 24 h after LPS injection for analysis of kidney pathologies, blood urea nitrogen(BUN) and creatinine levels; serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β) and IL-6 were detected by ELISA, and the expressions of Toll-like receptor-4(TLR4), MyD88, TRAF-6 and NF-κBp65 in the kidney were detected by Western blotting. Results Quercetin significantly lessened renal pathologies,lowered BUN and creatinine levels(P<0.05) and inhibited TNF-α, IL-1β, and IL-6 production in mice with LPS-induced AKI(P<0.05). Pretreatment with quercetin also significantly inhibited TLR4, MyD88, and TRAF-6 expressions and NF-κBp65 activation in the kidneys of the rats with LPS challenge(P<0.05). Conclusion Quercetin pretreatment can protect mice against LPSinduced AKI by inhibiting TLR4/NF-κB signaling pathway.
Objective To investigate the protective effect of quercetin against lipopolysaccharide(LPS)-induced acute kidney injury(AKI) in mice and explore its mechanism. Methods Forty male BALB/c mice were randomly divided into control group(with saline treatment), 15 mg/kg LPS group, and quercetin-treated groups with intragastric quercetin treatment(once daily for 3 consecutive days) at low(25 mg/kg) and high(50 mg/kg) dose prior to 15 mg/kg LPS injection. LPS was administered by intraperitoneally injection 1 after the last gavage of quercetin. The mice were sacrificed 24 h after LPS injection for analysis of kidney pathologies, blood urea nitrogen(BUN) and creatinine levels; serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β) and IL-6 were detected by ELISA, and the expressions of Toll-like receptor-4(TLR4), MyD88, TRAF-6 and NF-κBp65 in the kidney were detected by Western blotting. Results Quercetin significantly lessened renal pathologies,lowered BUN and creatinine levels(P<0.05) and inhibited TNF-α, IL-1β, and IL-6 production in mice with LPS-induced AKI(P<0.05). Pretreatment with quercetin also significantly inhibited TLR4, MyD88, and TRAF-6 expressions and NF-κBp65 activation in the kidneys of the rats with LPS challenge(P<0.05). Conclusion Quercetin pretreatment can protect mice against LPSinduced AKI by inhibiting TLR4/NF-κB signaling pathway.
引文
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[13]司海超,司小萌,刘展.槲皮素减轻坐骨神经慢性缩窄性损伤大鼠的神经病理性疼痛及其相关机制[J].第三军医大学学报, 2017, 39(1):54-9.
[14]梁青春,陈燕亭,李传翔,等.槲皮素调节ROS/TLR4信号通路抑制Ox-LDL诱导的血管平滑肌细胞钙化[J].南方医科大学学报, 2018,38(8):980-5.
[15]Fu H, Hu Z, Di X, et al.Tenuigenin exhibits protective effects against LPS-induced acute kidney injury via inhibiting TLR4/NF-κB signaling pathway[J]. Europ J Pharmacol, 2016, 791(6):229-34.
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[17]Bagshaw SM, George C, Bellomo R, et al. Changes in the incidence and outcome for early acute kidney injury in a cohort of Australian intensive care units[J]. Crit Care, 2007, 11(3):R68-80.
[18]Gaieski DF, Edwards JM, Kallan MJ, et al.Benchmarking the incidence and mortality of severe sepsis in the United States[J].Critical Care Med, 2013, 41(5):1167-74.
[19]Zarjou A, Agarwal A. Sepsis and acute kidney injury[J]. J Am Soc Nephrol, 2011, 22(6):999-1006.
[20]赵娜,田焕焕,李志,等.脓毒症并发急性肾损伤的危险因素分析与早期诊断[J].中华危重病急救医学, 2013, 25(9):542-5.
[21]Akcay A, Nguyen Q, Edelstein CL. Mediators of inflammation in acute kidney injury[J]. Mediators Inflamm, 2009, 16(8):137072-85.
[22]Huang RS, Zhou JJ, Feng YY, et al.Pharmacological inhibition of macrophage toll-like receptor 4/nuclear factor-kappa B alleviates rhabdomyolysis-induced acute kidney injury[J]. Chin Med J, 2017,130(18):2163-9.
[23]Zhao L, Xu L, Tao X, et al. Protective effect of the total flavonoids from rosa laevigata michx fruit on renal ischemia-reperfusion injury through suppression of oxidative stress and inflammation[J].Molecules, 2016, 21(7):185-96.
[24]Mir SM, Ravuri HG, Pradhan RK, et al. Ferulic acid protects lipopolysaccharide-induced acute kidney injury by suppressing inflammatory events and upregulating antioxidant defenses in Balb/c mice[J]. Biomed Pharmacoth, 2018, 100(3):304-15.
[25]Feng D, Wang Y, Liu Y, et al. DC-SIGN reacts with TLR-4 and regulates inflammatory cytokine expression via NF-κB activation in renal tubular epithelial cells during acute renal injury:DC-SIGN/TLR-4 regulates NF-κB activation[J]. Clin Experim Immunol, 2018,191(1):107-15.
[26]Zhang D, Li Y, Liu Y, et al.Paclitaxel ameliorates lipopolysaccharideinduced kidney injury by binding myeloid differentiation protein-2to block toll-like receptor 4-mediated nuclear factor-B activation and cytokine production[J]. J Pharmacol Experim Therapeut, 2013,345(1):69-75.
[27]Song J, Fan H, Li H, et al. Zingerone ameliorates lipopolysaccharideinduced acute kidney injury by inhibiting Toll-like receptor 4signaling pathway[J]. Europ J Pharmacol, 2016, 772(2):108-14.
[28]Fan HY, Qi D, Yu C, et al. Paeonol protects endotoxin-induced acute kidney injury:potential mechanism of inhibiting TLR4-NF-κB signal pathway[J]. Oncotarget, 2016, 7(26):39497-510.
[29]Beckmann DV, Carvalho FB, Mazzanti CM, et al.Neuroprotective role of quercetin in locomotor activities and cholinergic neurotransmission in rats experimentally demyelinated with ethidium bromide[J]. Life Sci, 2014, 103(2):79-87.
[30]檀昕,程安玮,孙金月,等.茶多酚和槲皮素对炎性脂肪细胞信号通路和炎性因子的影响[J].中国农学通报, 2018, 34(5):25-30.
[31]Ho AW, Wong CK, Lam CW. Tumor necrosis factor-alpha upregulates the expression of CCL2 and adhesion molecules of human proximal tubular epithelial cells through MAPK signaling pathways[J]. Immunobiology, 2008, 213(7):533-44.
[32]Leemans JC, Butter LM, Teske GD, et al.The toll interleukin-1receptor(IL-1R)8/single Ig domain IL-1R-related molecule modulates the renal response to bacterial infection[J]. Infec Immun,2012, 80(11):3812-20.
[33]Meng XM, Nikolic-Paterson DJ, Lan HY. Inflammatory processes in renal fibrosis[J]. Nat Rev Nephrol, 2014, 10(9):493-503.
[2] Alobaidi R, Basu RK, Goldstein SL, et al. Sepsis-associated acute kidney injury[J]. Semin Nephrol, 2015, 35(1):2-11.
[3] Zarbock A, Gomez H, Kellum JA. Sepsis-induced acute kidney injury revisited:pathophysiology, prevention and future therapies[J]. Curr Opin Crit Care, 2014, 20(6):588-95.
[4] Shi M, Zeng X, Guo F, et al. Anti-Inflammatory pyranochalcone derivative attenuates LPS-induced acute kidney injury via inhibiting TLR4/NF-κB pathway[J]. Molecules, 2017, 22(10):1683-95.
[5] Zhang L, Sun D, Bao Y, et al.Nerolidol protects against LPS-induced acute kidney injury via inhibiting TLR4/NF-κB signaling[J].Phytoth Res, 2017, 31(3):459-65.
[6] Ye HY, Jin J, Jin LW, et al. Chlorogenic acid attenuates lipopolysaccharide-induced acute kidney injury by inhibiting TLR4/NF-κB signal pathway[J]. Inflammation, 2017, 40(2):523-9.
[7] Zhou SJ, Wang G, Zhang WB. Effect of TLR4/MyD88 signaling pathway on sepsis-associated acute respiratory distress syndrome in rats, via regulation of macrophage activation and inflammatory response[J]. Exp Ther Med, 2018, 15(4):3376-84.
[8] Molteni M, Gemma S, Rossetti C. The role of Toll-Like receptor 4 in infectious and noninfectious inflammation[J]. Mediators Inflamm,2016, 20(11):936-49.
[9] Qu X, Qi D, Dong F, et al.Quercetin improves hypoxia-ischemia induced cognitive deficits via promoting remyelination in neonatal rat[J]. Brain Research, 2014, 1553(12):31-40.
[10]Wei X, Meng X, Yuan Y, et al. Quercetin exerts cardiovascular protective effects in LPS-induced dysfunction in vivo by regulating inflammatory cytokine expression, NF-κB phosphorylation and caspase activity[J]. Molecul Cellul Biochem, 2018, 32(4):1-10.
[11]Lu J, Zheng Y, Luo L, et al.Quercetin reverses d-galactose induced neurotoxicity in mouse brain[J]. Behav Brain Res, 2006, 171(2):251-60.
[12]李坚,张剑,董欣敏,等.槲皮素对内毒素性心肌损伤的保护作用及机制[J].南方医科大学学报, 2015, 35(7):1068-72.
[13]司海超,司小萌,刘展.槲皮素减轻坐骨神经慢性缩窄性损伤大鼠的神经病理性疼痛及其相关机制[J].第三军医大学学报, 2017, 39(1):54-9.
[14]梁青春,陈燕亭,李传翔,等.槲皮素调节ROS/TLR4信号通路抑制Ox-LDL诱导的血管平滑肌细胞钙化[J].南方医科大学学报, 2018,38(8):980-5.
[15]Fu H, Hu Z, Di X, et al.Tenuigenin exhibits protective effects against LPS-induced acute kidney injury via inhibiting TLR4/NF-κB signaling pathway[J]. Europ J Pharmacol, 2016, 791(6):229-34.
[16]Anderberg SB, Luther T, Frithiof R. Physiological aspects of Tolllike receptor 4 activation in sepsis-induced acute kidney injury[J].Acta Physiolog, 2017, 219(3):573-88.
[17]Bagshaw SM, George C, Bellomo R, et al. Changes in the incidence and outcome for early acute kidney injury in a cohort of Australian intensive care units[J]. Crit Care, 2007, 11(3):R68-80.
[18]Gaieski DF, Edwards JM, Kallan MJ, et al.Benchmarking the incidence and mortality of severe sepsis in the United States[J].Critical Care Med, 2013, 41(5):1167-74.
[19]Zarjou A, Agarwal A. Sepsis and acute kidney injury[J]. J Am Soc Nephrol, 2011, 22(6):999-1006.
[20]赵娜,田焕焕,李志,等.脓毒症并发急性肾损伤的危险因素分析与早期诊断[J].中华危重病急救医学, 2013, 25(9):542-5.
[21]Akcay A, Nguyen Q, Edelstein CL. Mediators of inflammation in acute kidney injury[J]. Mediators Inflamm, 2009, 16(8):137072-85.
[22]Huang RS, Zhou JJ, Feng YY, et al.Pharmacological inhibition of macrophage toll-like receptor 4/nuclear factor-kappa B alleviates rhabdomyolysis-induced acute kidney injury[J]. Chin Med J, 2017,130(18):2163-9.
[23]Zhao L, Xu L, Tao X, et al. Protective effect of the total flavonoids from rosa laevigata michx fruit on renal ischemia-reperfusion injury through suppression of oxidative stress and inflammation[J].Molecules, 2016, 21(7):185-96.
[24]Mir SM, Ravuri HG, Pradhan RK, et al. Ferulic acid protects lipopolysaccharide-induced acute kidney injury by suppressing inflammatory events and upregulating antioxidant defenses in Balb/c mice[J]. Biomed Pharmacoth, 2018, 100(3):304-15.
[25]Feng D, Wang Y, Liu Y, et al. DC-SIGN reacts with TLR-4 and regulates inflammatory cytokine expression via NF-κB activation in renal tubular epithelial cells during acute renal injury:DC-SIGN/TLR-4 regulates NF-κB activation[J]. Clin Experim Immunol, 2018,191(1):107-15.
[26]Zhang D, Li Y, Liu Y, et al.Paclitaxel ameliorates lipopolysaccharideinduced kidney injury by binding myeloid differentiation protein-2to block toll-like receptor 4-mediated nuclear factor-B activation and cytokine production[J]. J Pharmacol Experim Therapeut, 2013,345(1):69-75.
[27]Song J, Fan H, Li H, et al. Zingerone ameliorates lipopolysaccharideinduced acute kidney injury by inhibiting Toll-like receptor 4signaling pathway[J]. Europ J Pharmacol, 2016, 772(2):108-14.
[28]Fan HY, Qi D, Yu C, et al. Paeonol protects endotoxin-induced acute kidney injury:potential mechanism of inhibiting TLR4-NF-κB signal pathway[J]. Oncotarget, 2016, 7(26):39497-510.
[29]Beckmann DV, Carvalho FB, Mazzanti CM, et al.Neuroprotective role of quercetin in locomotor activities and cholinergic neurotransmission in rats experimentally demyelinated with ethidium bromide[J]. Life Sci, 2014, 103(2):79-87.
[30]檀昕,程安玮,孙金月,等.茶多酚和槲皮素对炎性脂肪细胞信号通路和炎性因子的影响[J].中国农学通报, 2018, 34(5):25-30.
[31]Ho AW, Wong CK, Lam CW. Tumor necrosis factor-alpha upregulates the expression of CCL2 and adhesion molecules of human proximal tubular epithelial cells through MAPK signaling pathways[J]. Immunobiology, 2008, 213(7):533-44.
[32]Leemans JC, Butter LM, Teske GD, et al.The toll interleukin-1receptor(IL-1R)8/single Ig domain IL-1R-related molecule modulates the renal response to bacterial infection[J]. Infec Immun,2012, 80(11):3812-20.
[33]Meng XM, Nikolic-Paterson DJ, Lan HY. Inflammatory processes in renal fibrosis[J]. Nat Rev Nephrol, 2014, 10(9):493-503.