荷叶总生物碱激活肝脏AMPK/Nrf2通路缓解对乙酰氨基酚诱导的小鼠急性肝损伤
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摘要
为探讨荷叶总生物碱(TAL)对对乙酰氨基酚(APAP)所致小鼠急性肝损伤的保护作用及可能机制,小鼠被随机分为正常组,模型组和TAL低、高剂量组(30、100 mg/kg),连续灌胃给药七天后,除正常组外,其余各组腹腔注射300 mg/kg的APAP诱导急性肝损伤。12小时后,收集各组小鼠的血清与肝脏样本,进行后续实验。结果显示,与模型组相比,TAL可显著降低血清中的ALT和AST活性,下调肝组织中TNF-α、IL-1β、IL-6和MDA含量,上调肝组织中SOD、CAT、GSH-Px和GSH的水平。此外,TAL还明显改善了APAP诱导的小鼠肝组织病变。在TAL的作用下,肝内AMPK磷酸化水平提高,Nrf2蛋白入核,HO-1和GCLC基因表达上调。综上所述,TAL对APAP诱导的急性肝损伤具有保护作用,其机制可能与激活AMPK/Nrf2通路相关。
This study aimed to understand the protective effects of total alkaloids from lotus leaves(TAL) on acute liver injury induced by acetaminophen(APAP) in mice and the potential mechanisms. Mice were randomly divided into 4 groups,i.e.,the normal group,the model group,the TAL low-dosage group(30 mg/kg) and the TAL high-dosage group(100 mg/kg). Animals received TAL through gavage for 7 consecutive days. Then,except the normal group,other groups received APAP(300 mg/kg) by intraperitoneal injection to provoke acute liver injury. Serum and livers of mice were collected 12 h later and subjected to further experiments. Our results showed that as compared to the model group,TAL significantly decreased activities of ALT and AST in the serum and contents of TNF-α,IL-1β,IL-6 and MDA in the liver. In addition,TAL dose-dependently elevated hepatic levels of SOD,CAT,GSH-Px and GSH. Moreover,APAP-induced liver histopathological changes were dramatically alleviated by TAL. Mechanistically,TAL considerably boosted AMPK phosphorylation,induced nuclear translocation of Nrf2 and promoted expression of HO-1 and GCLC. In conclusion,this study revealed that TAL has the capacity of ameliorating APAP-induced acute liver injury,and the underlying mechanisms are related with the activation of hepatic AMPK/Nrf2 cascade.
This study aimed to understand the protective effects of total alkaloids from lotus leaves(TAL) on acute liver injury induced by acetaminophen(APAP) in mice and the potential mechanisms. Mice were randomly divided into 4 groups,i.e.,the normal group,the model group,the TAL low-dosage group(30 mg/kg) and the TAL high-dosage group(100 mg/kg). Animals received TAL through gavage for 7 consecutive days. Then,except the normal group,other groups received APAP(300 mg/kg) by intraperitoneal injection to provoke acute liver injury. Serum and livers of mice were collected 12 h later and subjected to further experiments. Our results showed that as compared to the model group,TAL significantly decreased activities of ALT and AST in the serum and contents of TNF-α,IL-1β,IL-6 and MDA in the liver. In addition,TAL dose-dependently elevated hepatic levels of SOD,CAT,GSH-Px and GSH. Moreover,APAP-induced liver histopathological changes were dramatically alleviated by TAL. Mechanistically,TAL considerably boosted AMPK phosphorylation,induced nuclear translocation of Nrf2 and promoted expression of HO-1 and GCLC. In conclusion,this study revealed that TAL has the capacity of ameliorating APAP-induced acute liver injury,and the underlying mechanisms are related with the activation of hepatic AMPK/Nrf2 cascade.
引文
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13 Zhang L,Hao J,Huang X,et al.Total alkaloids from the seed embryo of nelumbo nucifera ameliorate inflammation in the livers of mice with lipopolysaccharide/D-galactosamine-induced acute liver failure[J].Food Sci(食品科学),2018,39:220-225.
14 Zhou JG.Study of hepatoprotective effect of the total alkaloids from Corydalis saxicola bunting of the acute chemical hepatic injury model mice[J].Chin Modern Med(中国当代医药),2010,17(27):29-30.
15 Truong VL,Bak MJ,Jun M,et al.Antioxidant defense and hepatoprotection by procyanidins from almond(Prunus amygdalus)skins[J].J Agric Food Chem,2014,62:8668.
16 Yan H,Huang Z,Bai Q,et al.Natural product andrographolide alleviated APAP-induced liver fibrosis by activating Nrf2antioxidant pathway[J].Toxicology,2018,396-397:1-12.
17 Lin LC,Zhao FF,Wang M,et al.Protection of auriculatone sulphate against APAP-induced acute liver injury in mice[J].Nat Prod Res Dev(天然产物研究与开发),2018,30:761-766.
18 Bae GH,Lee SK,Kim HS,et al.Lysophosphatidic acid protects against acetaminophen-induced acute liver injury[J].Exp Mol Med,2017,49:e407.
19 Joo MS,Kim WD,Lee KY,et al.AMPK facilitates nuclear accumulation of Nrf2 by phosphorylating at serine 550[J].Mol Cell Biol,2016,36:1931.
20 Jain AK,Jaiswal AK.GSK-3 acts upstream of fyn kinase in regulation of nuclear export and degradation of NF-E2 related factor 2[J].J Biol Chem,2007,282:16502.
21 Son YO,Pratheeshkumar P,Roy RV,et al.Nrf2/p62 signaling in apoptosis resistance and its role in cadmium-induced carcinogenesis[J].J Biol Chem,2014,289:28660-28675.
22 Lee DH,Han DH,Nam KT,et al.Ezetimibe,an NPC1L1 inhibitor,is a potent Nrf2 activator that protects mice from dietinduced nonalcoholic steatohepatitis[J].Free Radic Biol Med,2016,99:520-532.
23 Wei M,Zheng Z,Shi L,et al.Natural polyphenol chlorogenic acid protects against acetaminophen-induced hepatotoxicity by activating ERK/Nrf2 antioxidative pathway[J].Toxicol Sci,2017,162:99-112.
24 Monteiro LOF,Fernandes RS,Oda CMR,et al.Protective effect of dioscin against thioacetamide-induced acute liver injury via FXR/AMPK signaling pathway[J].Biomed Pharmacother,2018,97:481-488.
25 Wang L,Zhang S,Cheng H,et al.Nrf2-mediated liver protection by esculentoside A against acetaminophen toxicity through the AMPK/Akt/GSK3βpathway[J].Free Radic Biol Med,2016,101:401-412.
2 Karthivashan G,Arulselvan P,Tan SW,et al.The molecular mechanism underlying the hepatoprotective potential of Moringa oleifera,leaves extract against acetaminophen induced hepatotoxicity in mice[J].J Funct Foods,2015,17:115-126.
3 Wu H,Zhang G,Huang L,et al.Hepatoprotective effect of polyphenol-enriched fraction from folium microcos on oxidative stress and apoptosis in acetaminophen-induced liver injury in mice[J].Oxid Med Cell Longev,2017,4:3631565.
4 Lv H,Xiao Q,Zhou J,et al.Licochalcone A upregulates Nrf2antioxidant pathway and thereby alleviates acetaminophen-induced hepatotoxicity[J].Front Pharmacol,2018,9:147.
5 Hu LF,Wang Y,Ren RJ,et al.Anti-oxidative stress actions and regulation mechanisms of Keap1-Nrf2/ARE signal pathway[J].J Int Pharm Res(国际药学研究杂志),2016,43:146-152.
6 Huang W,Wang Y,Jiang X,et al.Protective effect of flavonoids from Ziziphus jujuba cv.Jinsixiaozao against acetaminophen-induced liver injury by inhibiting oxidative stress and inflammation in mice[J].Molecules,2017,22:3887-3893.
7 Kouam AF,Yuan F,Njayou FN,et al.Induction of Mkp-1and nuclear translocation of Nrf2 by limonoids from khaya grandifoliola C.DC protect L-02 hepatocytes against acetaminophen-induced hepatotoxicity[J].Front Pharmacol,2017,8:653.
8 Xia N,Chen YL,Tao HY,et al.Protective effect of polysaccharides from Morus nigra Linn.fruits on CCl4-induced liver damage in rats[J].Food Sci(食品科学),2015,36:247-251.
9 Xin Z,Deng Q,Chen H,et al.Characterizations and hepatoprotective effect of polysaccharides from Mori Fructus,in rats with alcoholic-induced liver injury[J].Int J Biol Macromol,2017,102:60-67.
10 Liu BT.The chemical composition and pharmacological effects of nelumbo nucifera leaves[J].Contemp Chem Ind(当代化工研究),2016,7:134-135.
11 Xiao GQ,Tian Y,Lu XY.Study on scavenging oxygen free radicals capabilities of total alkaloids from lotus leaves[J].Chem&Bioeng(化学与生物工程),2011,28(10):35-38.
12 He XX.Study on analysis and pharmacokinetics of alkaloids from nelumbo nucifera leaves[D].Tianjin:Tianjin University of Science&Technology(天津科技大学),2013.
13 Zhang L,Hao J,Huang X,et al.Total alkaloids from the seed embryo of nelumbo nucifera ameliorate inflammation in the livers of mice with lipopolysaccharide/D-galactosamine-induced acute liver failure[J].Food Sci(食品科学),2018,39:220-225.
14 Zhou JG.Study of hepatoprotective effect of the total alkaloids from Corydalis saxicola bunting of the acute chemical hepatic injury model mice[J].Chin Modern Med(中国当代医药),2010,17(27):29-30.
15 Truong VL,Bak MJ,Jun M,et al.Antioxidant defense and hepatoprotection by procyanidins from almond(Prunus amygdalus)skins[J].J Agric Food Chem,2014,62:8668.
16 Yan H,Huang Z,Bai Q,et al.Natural product andrographolide alleviated APAP-induced liver fibrosis by activating Nrf2antioxidant pathway[J].Toxicology,2018,396-397:1-12.
17 Lin LC,Zhao FF,Wang M,et al.Protection of auriculatone sulphate against APAP-induced acute liver injury in mice[J].Nat Prod Res Dev(天然产物研究与开发),2018,30:761-766.
18 Bae GH,Lee SK,Kim HS,et al.Lysophosphatidic acid protects against acetaminophen-induced acute liver injury[J].Exp Mol Med,2017,49:e407.
19 Joo MS,Kim WD,Lee KY,et al.AMPK facilitates nuclear accumulation of Nrf2 by phosphorylating at serine 550[J].Mol Cell Biol,2016,36:1931.
20 Jain AK,Jaiswal AK.GSK-3 acts upstream of fyn kinase in regulation of nuclear export and degradation of NF-E2 related factor 2[J].J Biol Chem,2007,282:16502.
21 Son YO,Pratheeshkumar P,Roy RV,et al.Nrf2/p62 signaling in apoptosis resistance and its role in cadmium-induced carcinogenesis[J].J Biol Chem,2014,289:28660-28675.
22 Lee DH,Han DH,Nam KT,et al.Ezetimibe,an NPC1L1 inhibitor,is a potent Nrf2 activator that protects mice from dietinduced nonalcoholic steatohepatitis[J].Free Radic Biol Med,2016,99:520-532.
23 Wei M,Zheng Z,Shi L,et al.Natural polyphenol chlorogenic acid protects against acetaminophen-induced hepatotoxicity by activating ERK/Nrf2 antioxidative pathway[J].Toxicol Sci,2017,162:99-112.
24 Monteiro LOF,Fernandes RS,Oda CMR,et al.Protective effect of dioscin against thioacetamide-induced acute liver injury via FXR/AMPK signaling pathway[J].Biomed Pharmacother,2018,97:481-488.
25 Wang L,Zhang S,Cheng H,et al.Nrf2-mediated liver protection by esculentoside A against acetaminophen toxicity through the AMPK/Akt/GSK3βpathway[J].Free Radic Biol Med,2016,101:401-412.