基于Lable-free蛋白质组学技术研究大黄素致大鼠肝损伤作用机制
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摘要
目的:探索连续4周服用大黄素对大鼠肝组织蛋白表达的影响。方法:选取以大黄折算临床剂量100倍的大黄素灌胃SD大鼠4周(HG1,HG2,HG3,HG4)为模型,采用Lable-free蛋白质组学方法检测肝组织蛋白表达,并对所得结果进行分析。结果:通过筛选,HG1、HG2、HG3、HG4组中分别有158个、650个、219个、378个差异蛋白,这些差异蛋白经GO分析发现主要与细胞和代谢过程相关。其中,4组中共有差异蛋白25个,GO分类表明25个蛋白中有10个为与催化活性相关的蛋白。大黄素的存在使α-2-HS-糖蛋白(热稳定性糖蛋白)表达上调,该蛋白的浓度与肝病的严重程度正相关;并且组织蛋白酶家族蛋—富含半胱氨酸的蛋白酶表达下调。结论:长期服用大黄素会增加肝损伤的概率,其可能是由于含半胱氨酸蛋白酶的表达下调,使组织功能出现紊乱,导致肝损伤的出现,并且这一结果在低剂量组中也得到了印证。
Objective: To investigate the influence of 4-week's emodin on protein expression of rat's liver tissue. Methods: SD rats were given emodin 100 times more than the clinical dose of Rhubarb for 4 weeks and were divided into four groups( HG1,HG2,HG3,HG4). Protein expression in liver tissue was detected by lable-free proteomic technology. Results: By screening,there were158,650,219,and 378 differential proteins in HG1,HG2,HG3,and HG4 group respectively,which were found to be mainly associated with cellular and metabolic processes by GO analysis. Among them,there were 25 differential proteins in the four groups,and GO classification indicated that 10 of the 25 proteins were proteins related to catalytic activity. α-2-HS-glycoprotein( thermostable glycoprotein) expression was upregulated by the presence of emodin,and the concentration of this protein positively correlated with the severity of liver disease; and cathepsin family egg-cysteine-rich protease expression was downregulated. Conclusion: Long-term administration of emodin increases the probability of liver injury,which may be due to the down-regulation of the expression of cysteine-containing proteases,which leads to the disorder of tissue function and leads to the appearance of liver injury,and this result has also been confirmed in the low-dose group.
Objective: To investigate the influence of 4-week's emodin on protein expression of rat's liver tissue. Methods: SD rats were given emodin 100 times more than the clinical dose of Rhubarb for 4 weeks and were divided into four groups( HG1,HG2,HG3,HG4). Protein expression in liver tissue was detected by lable-free proteomic technology. Results: By screening,there were158,650,219,and 378 differential proteins in HG1,HG2,HG3,and HG4 group respectively,which were found to be mainly associated with cellular and metabolic processes by GO analysis. Among them,there were 25 differential proteins in the four groups,and GO classification indicated that 10 of the 25 proteins were proteins related to catalytic activity. α-2-HS-glycoprotein( thermostable glycoprotein) expression was upregulated by the presence of emodin,and the concentration of this protein positively correlated with the severity of liver disease; and cathepsin family egg-cysteine-rich protease expression was downregulated. Conclusion: Long-term administration of emodin increases the probability of liver injury,which may be due to the down-regulation of the expression of cysteine-containing proteases,which leads to the disorder of tissue function and leads to the appearance of liver injury,and this result has also been confirmed in the low-dose group.
引文
[1]国家药典委员会.中华人民共和国药典[S].一部.北京:中国医药科技出版社,2015:22-23.
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[8]张瑞晨,刘斌,孙震晓,等.何首乌提取物对人正常肝细胞L02周期阻滞及凋亡的影响[J].中西医结合学报,2010,8(6):554-561.
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[11]Vargas F,Fraile G,Velásquez M,et al.Studies on the photostability and phototoxicity of aloe-emodin,emodin and rhein[J].Pharmazie,2002,57(6):399-404.
[12]Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW264.7 macrophages[J].Int J Mol Med,2005,16(1):41-47.
[13]Wu Y,Tu X,Lin G,et al.Emodin-mediated protection from acute myocardial infarction via inhibition of inflammation and apoptosis in local ischemic myocardium[J].Life Sci,2007,81(17-18):1332-1338.
[14]Lu Y,Zhang J,Qian J.The effect of emodin on VEGF receptors in human colon cancer cells[J].Cancer Biother Radiopharm,2008,23(2):222-228.
[15]He ZH,He MF,Ma SC,et al.Anti-angiogenic effects of rhubarb and its anthraquinone derivatives[J].J Ethnopharmacol,2009,121(2):313-317.
[16]Huang Q,Shen HM,Ong CN.Inhibitory effect of emodin on tumor invasion through suppression of activator protein-1 and nuclear factor-kappaB[J].Biochem Pharmacol,2004,68(2):361-371.
[17]Huang Q,Shen HM,Shui G,et al.Emodin inhibits tumor cell adhesion through disruption of the membrane lipid Raft-associated integrin signaling pathway[J].Cancer Res,2006,66(11):5807-5815.
[18]Huang Q,Lu G,Shen HM,et al.Anti-cancer properties of anthraquinones from rhubarb[J].Med Res Rev,2007,27(5):609-630.
[19]Srinivas G,Babykutty S,Sathiadevan PP,et al.Molecular mechanism of emodin action:transition from laxative ingredient to an antitumor agent[J].Med Res Rev,2007,27(5):591-608.
[20]Su YT,Chang HL,Shyue SK,et al.Emodin induces apoptosis in human lung adenocarcinoma cells through a reactive oxygenspecies-dependent mitochondrial signaling pathway[J].Biochem Pharmacol,2005,70(2):229-241.
[21]Kalabay L,Jakab L,Prohászka Z,et al.Human fetuin/alpha2HS-glycoprotein level as a novel indicator of liver cell function and shortterm mortality in patients with liver cirrhosis and liver cancer[J].Eur J Gastroenterol Hepatol,2002,14(4):389-394.
[22]Wang C,Dai X,Liu H,et al.Involvement of PPARγin emodin-induced HK-2 cell apoptosis[J].Toxicol In Vitro,2015,29(1):228-233.
[2]Cho HC,Min HJ,Ha CY,et al.Reactivation of Pulmonary Tuberculosis in a Patient with Polygonum multiflorum Thunb-Induced Hepatitis[J].Gut Liver,2009,3(1):52-56.
[3]Jung KA,Min HJ,Yoo SS,et al.Drug-Induced Liver Injury:Twenty Five Cases of Acute Hepatitis Following Ingestion of Polygonum multiflorum Thunb[J].Gut Liver,2011,5(4):493-499.
[4]Dong H,Slain D,Cheng J,et al.Eighteen cases of liver injury following ingestion of Polygonum multiflorum[J].Complement Ther Med,2014,22(1):70-74.
[5]Huang Q,Lu G,Shen HM,et al.Anti-cancer properties of anthraquinones from rhubarb[J].Med Res Rev,2007,27(5):609-630.
[6]Pecere T,Gazzola MV,Mucignat C,et al.Aloe-emodin is a new type of anticancer agent with selective activity against neuroectodermal tumors[J].Cancer Res,2000,60(11):2800-2804.
[7]National Toxicology Program.NTP Toxicology and Carcinogenesis Studies of EMODIN(CAS NO.518-82-1)Feed Studies in F344/NRats and B6C3F1 Mice[J].Natl Toxicol Program Tech Rep Ser,2001,493:1-278.
[8]张瑞晨,刘斌,孙震晓,等.何首乌提取物对人正常肝细胞L02周期阻滞及凋亡的影响[J].中西医结合学报,2010,8(6):554-561.
[9]杨波.何首乌临床应用中的不良反应分析[J].医学信息,2015,28(6):200-201.
[10]Arosio B,Gagliano N,Fusaro LM,Aloe-Emodin quinone pretreatment reduces acute liver injury induced by carbon tetrachloride[J].Pharmacol Toxicol,2000,87(5):229-33.
[11]Vargas F,Fraile G,Velásquez M,et al.Studies on the photostability and phototoxicity of aloe-emodin,emodin and rhein[J].Pharmazie,2002,57(6):399-404.
[12]Li HL,Chen HL,Li H,et al.Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW264.7 macrophages[J].Int J Mol Med,2005,16(1):41-47.
[13]Wu Y,Tu X,Lin G,et al.Emodin-mediated protection from acute myocardial infarction via inhibition of inflammation and apoptosis in local ischemic myocardium[J].Life Sci,2007,81(17-18):1332-1338.
[14]Lu Y,Zhang J,Qian J.The effect of emodin on VEGF receptors in human colon cancer cells[J].Cancer Biother Radiopharm,2008,23(2):222-228.
[15]He ZH,He MF,Ma SC,et al.Anti-angiogenic effects of rhubarb and its anthraquinone derivatives[J].J Ethnopharmacol,2009,121(2):313-317.
[16]Huang Q,Shen HM,Ong CN.Inhibitory effect of emodin on tumor invasion through suppression of activator protein-1 and nuclear factor-kappaB[J].Biochem Pharmacol,2004,68(2):361-371.
[17]Huang Q,Shen HM,Shui G,et al.Emodin inhibits tumor cell adhesion through disruption of the membrane lipid Raft-associated integrin signaling pathway[J].Cancer Res,2006,66(11):5807-5815.
[18]Huang Q,Lu G,Shen HM,et al.Anti-cancer properties of anthraquinones from rhubarb[J].Med Res Rev,2007,27(5):609-630.
[19]Srinivas G,Babykutty S,Sathiadevan PP,et al.Molecular mechanism of emodin action:transition from laxative ingredient to an antitumor agent[J].Med Res Rev,2007,27(5):591-608.
[20]Su YT,Chang HL,Shyue SK,et al.Emodin induces apoptosis in human lung adenocarcinoma cells through a reactive oxygenspecies-dependent mitochondrial signaling pathway[J].Biochem Pharmacol,2005,70(2):229-241.
[21]Kalabay L,Jakab L,Prohászka Z,et al.Human fetuin/alpha2HS-glycoprotein level as a novel indicator of liver cell function and shortterm mortality in patients with liver cirrhosis and liver cancer[J].Eur J Gastroenterol Hepatol,2002,14(4):389-394.
[22]Wang C,Dai X,Liu H,et al.Involvement of PPARγin emodin-induced HK-2 cell apoptosis[J].Toxicol In Vitro,2015,29(1):228-233.