山柰酚抵抗对乙酰氨基酚引起的肝细胞损伤研究
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摘要
目的:探究山柰酚(KA)抵抗对乙酰氨基酚(APAP)诱导的肝细胞损伤的作用。方法:CCK-8法检测KA对肝细胞活性的影响,检测细胞LDH释放比率确定APAP诱导肝细胞损伤模型的浓度和时间; CCK-8法检测肝细胞活性,检测肝细胞释放的LDH,观察肝细胞形态评估KA对对乙酰氨基酚诱导肝细胞损伤的影响; Western Blot检测p-JNK、JNK、Endo G蛋白表达。结果:本研究采用5 mmol/L APAP诱导肝细胞24 h建立药物肝损伤细胞模型。与APAP组比较,KA组肝细胞细胞的形态明显改善,凋亡细胞明显减少(P <0. 01),LDH释放比率明显下降(P <0. 001),Western Blot结果表明经KA治疗后,肝p-JNK和Endo G蛋白表达明显下调。结论:该研究表明KA通过抑制p-JNK和Endo G来抵抗APAP诱导肝细胞凋亡以达到肝细胞的保护作用。
Objective: To investigate the protective effects of kaempferol against acetaminophen in hepatocytes-induced injury.Methods: Cholecystokinin-8 (CCK-8) was used to detect the vitality of hepatocytes treated by kaempferol. The lactate dehydrogenase (LDH) released ratio was detected,which could determine the concentration and time of acetaminophen (APAP)-induced hepatocytes injury model. CCK-8 and LDH released ratio were used to detect the vitality of hepatocytes treated by APAP and kaempferol. In the meanwhile,hepatocyte morphology was to be record. P-Jun N-terminal kinase (P-JNK),Jun N-terminal kinase (JNK),Endo G protein expression were detected by Western Blot searching. Results: The APAP concentration was determined to be 5 mmol/L and the time was 24 hours to induce liver injury cell with drug. Compared with the APAP group,the morphology of hepatocytes was significantly improved. The ratio of apoptotic cells decreased significantly (P < 0. 01),and the ratio of released LDH content decreased significantly (P < 0. 001). Western Blot showed that P-JNK and Endo G were significantly down-regulated. Conclusion: This experiment demonstrates that kaempferol has a protective effect on APAP-induced hepatocytes injury.
Objective: To investigate the protective effects of kaempferol against acetaminophen in hepatocytes-induced injury.Methods: Cholecystokinin-8 (CCK-8) was used to detect the vitality of hepatocytes treated by kaempferol. The lactate dehydrogenase (LDH) released ratio was detected,which could determine the concentration and time of acetaminophen (APAP)-induced hepatocytes injury model. CCK-8 and LDH released ratio were used to detect the vitality of hepatocytes treated by APAP and kaempferol. In the meanwhile,hepatocyte morphology was to be record. P-Jun N-terminal kinase (P-JNK),Jun N-terminal kinase (JNK),Endo G protein expression were detected by Western Blot searching. Results: The APAP concentration was determined to be 5 mmol/L and the time was 24 hours to induce liver injury cell with drug. Compared with the APAP group,the morphology of hepatocytes was significantly improved. The ratio of apoptotic cells decreased significantly (P < 0. 01),and the ratio of released LDH content decreased significantly (P < 0. 001). Western Blot showed that P-JNK and Endo G were significantly down-regulated. Conclusion: This experiment demonstrates that kaempferol has a protective effect on APAP-induced hepatocytes injury.
引文
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[13]贺兰芝,孟雅坤,韩延忠,等.木犀草素对对乙酰氨基酚诱导的L02肝细胞损伤的保护作用[J].中国中药杂志,2016,41(22):4234-4239.
[14]Kunimaro Furuta,Yuichi Yoshida,Satoshi Ogura,et al.Gab1 Adaptor Protein Acts As a Gatekeeper to Balance Hepatocyte Death and Proliferation During Acetaminophen-Induced Liver Injury in Mice[J].Hepatology,2016,63(4):1340-1355.
[15]Mingzhu Yan,Yazhen Huo,Shutao Yin,et al.Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions[J].Redox Biology,2017,17:274-283.
[2]Jaeschke H,Knight TR,Bajt ML.The role of oxidant stress and reactive nitrogen species in acetaminophen hepatotoxicity[J].Toxicol Lett2003,144(3):279-288.
[3]Mc Gill MR,Sharpe MR,Williams CD,et al.The mechanism underlying acetaminophen-induced hepatotoxicity in humans and mice involves mitochondrial damage and nuclear DNA fragmentation[J].JClin Invest 2012,122(4):1574-1583.
[4]Oliver Krenkel,Jana C.Mossanen,et al.Immune mechanisms in acetaminophen-induced acute liver failure[J].Hepatobiliary Surg Nutr2014,3(6):331-343.
[5]Adebayo D,Mookerjee RP and Jalan R.Mechanistic biomarkers in acute liver injury:are we there yet[J].J Hepatol 2012,56(5):1003-1005.
[6]Calderon-Montano JM,Burgos-Moron E,Perez-Guerrero C,et al.A review on the dietary flavonoid kaempferol[J].Mini Rev Med Chem,2011,11(4):298-344.
[7]汤利华,方超,王浩然,等.山奈酚对高糖诱导的糖尿病肾病大鼠肾功能和组织病理损伤的保护作用[J].免疫学杂志,2018,34(12):1041-1046.
[8]康桂兰,景增秀.山奈酚通过调控AMPK/Nrf2/HO-1信号通路缓解ox-LDL介导的内皮细胞损伤[J].中国免疫学杂志,2018,34(4):525-530.
[9]Qi-Sheng Yang,Li-Ping He,Xian-Long Zhou,et al.Kaempferol pretreatment modulates systemic inflammation and oxidative stress following hemorrhagic shock in mice[J].Chinese Medicine,2015,10(1):6-13.
[10]Wenzhen Liao,Luying Chen,Xiang,et al.Protective effects of kaempferol against reactive oxygen species-induced hemolysis and its antiproliferative activity on human cancer cells[J].European Journal of Medicinal Chemistry,2016,114:24-32.
[11]Seung-Hee Kim,Kyung-A Hwang,Kyung-Chul Choi.Treatment with kaempferol suppresses breast cancer cell growth caused by estrogen and triclosan in cellular and xenograft breast cancer models[J].The Journal of Nutritional Biochemistry,2016,28:70-82.
[12]仇炜,林俊,张磊,等.山奈酚抑制顺铂诱导的心肌细胞凋亡[J].中国分子心脏病学杂志,2017,17(1):1992-1995.
[13]贺兰芝,孟雅坤,韩延忠,等.木犀草素对对乙酰氨基酚诱导的L02肝细胞损伤的保护作用[J].中国中药杂志,2016,41(22):4234-4239.
[14]Kunimaro Furuta,Yuichi Yoshida,Satoshi Ogura,et al.Gab1 Adaptor Protein Acts As a Gatekeeper to Balance Hepatocyte Death and Proliferation During Acetaminophen-Induced Liver Injury in Mice[J].Hepatology,2016,63(4):1340-1355.
[15]Mingzhu Yan,Yazhen Huo,Shutao Yin,et al.Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions[J].Redox Biology,2017,17:274-283.