老鼠簕生物碱A对肝纤维化大鼠PI3K/Akt/m TOR/p70S6K信号通路的影响
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摘要
目的观察老鼠簕生物碱A(HBOA)对四氯化碳(CCl4)致肝纤维化大鼠磷酸肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(m TOR)/p70核糖体蛋白S6激酶(p70S6K)信号通路的影响,探讨HBOA抗肝纤维化的作用机制。方法大鼠随机分成对照组、模型组及HBOA高、中、低剂量(100、50、25 mg/kg)组和秋水仙碱(0.4 mg/kg)组。除对照组外,其余各组ig给予50%CCl4橄榄油溶液,每周2次,连续12周,诱导肝纤维化大鼠模型。于造模第9周起,给药组分别ig相应的受试药物,每天1次,给药4周。实验结束后,计算各组大鼠体质量变化、肝脏指数;检测各组大鼠肝匀浆中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活性;蛋白免疫印迹法(Western blotting)法检测肝组织P-PI3K、p-Akt、p-mTOR、p-p70S6K蛋白的表达。结果与模型组比较,各给药组大鼠体质量增加量显著升高,均能降低肝脏指数及肝组织中ALT、AST活性;此外,HBOA高、中剂量组均能显著抑制p-PI3K、p-Akt、p-m TOR、p-p70S6K蛋白的表达。结论 HBOA对肝纤维化大鼠具有一定的保护作用,其机制可能与抑制PI3K/Akt/m TOR/p70S6K信号通路有关。
Objective To observe the effect of Acanthus ilicifolius alkaloid A(HBOA) on PI3 K/AKT/m TOR/p70S6K signaling pathway in rats with hepatic fibrosis induced by carbon tetrachloride(CCl4),and to explore the mechanism of action of HBOA against liver fibrosis.Methods Rats were randomly divided into normal group,model group,high,medium;and low-dose HBOA groups(100,50,25 mg/kg),and colchicine group(0.4 mg/kg).Except for the normal group,the rats in other groups were given with a 50% CCl4 olive oil solution twice a week for 12 weeks to induce a rat model of liver fibrosis.From the ninth week of modeling,the drug-administered group was given the corresponding test drug once daily for 4 weeks.After the experiment,the body mass change and liver index were calculated.The levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the liver homogenate of each group were detected.The protein expression of p-PI3 K,p-Akt,p-mTOR,and p-p70S6K in liver tissue was detected by Western blotting.Results Compared with the model group,the body weight of mice of each drug-administered group was significantly increased,and the liver index,and ALT and AST levels were decreased in liver tissue.In addition,HBOA high and medium-dose groups significantly inhibited the protein expression of p-PI3K,p-Akt,p-mTOR,and p-p70S6K.Conclusion HBOA has a protective effect on hepatic fibrosis rats,and its mechanism may be related to the inhibition of PI3K/Akt/mTOR/p70S6K signaling pathway.
Objective To observe the effect of Acanthus ilicifolius alkaloid A(HBOA) on PI3 K/AKT/m TOR/p70S6K signaling pathway in rats with hepatic fibrosis induced by carbon tetrachloride(CCl4),and to explore the mechanism of action of HBOA against liver fibrosis.Methods Rats were randomly divided into normal group,model group,high,medium;and low-dose HBOA groups(100,50,25 mg/kg),and colchicine group(0.4 mg/kg).Except for the normal group,the rats in other groups were given with a 50% CCl4 olive oil solution twice a week for 12 weeks to induce a rat model of liver fibrosis.From the ninth week of modeling,the drug-administered group was given the corresponding test drug once daily for 4 weeks.After the experiment,the body mass change and liver index were calculated.The levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the liver homogenate of each group were detected.The protein expression of p-PI3 K,p-Akt,p-mTOR,and p-p70S6K in liver tissue was detected by Western blotting.Results Compared with the model group,the body weight of mice of each drug-administered group was significantly increased,and the liver index,and ALT and AST levels were decreased in liver tissue.In addition,HBOA high and medium-dose groups significantly inhibited the protein expression of p-PI3K,p-Akt,p-mTOR,and p-p70S6K.Conclusion HBOA has a protective effect on hepatic fibrosis rats,and its mechanism may be related to the inhibition of PI3K/Akt/mTOR/p70S6K signaling pathway.
引文
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[17]Yang L,Zhao Z X,Hou J L,et al.Hepatic expression of CB1 in rats with fibrosis and the relationship with FAK[J].J Clin Hepatol,2011,27(8):824-826.
[18]Piguet A C,Dufour J F.PI3K/PTEN/Akt pathway[J].JHepatol,2011,54(6):1317-1319.
[19]G?bele E,Reif S,Tsukada S,et al.The role of p70S6Kinhepatic stellate cell collagen gene expression and cell proliferation[J].J Biol Chem,2005,280(14):13374-13382.
[20]Wei L,Chen Q,Guo A,et al.Asiatic acid attenuates CCl-induced liver fibrosis in rats by regulating the PI3K/AKT/m TOR and Bcl-2/Bax signaling pathways[J].Inter Immunopharmacol,2018,doi:10.1016/j.intimp.2018.04.016.
[21]占书箱,黄成,马陶陶,等.抑制p70S6K对HSC-T6增殖活化的作用研究[J].安徽医科大学学报,2014,49(8):1062-1067.
[22]张坤,姜妙娜,张彩华,等.中药肝复康对肝星状细胞m TOR/P70S6K信号通路的干预作用[J].实用肝脏病杂志,2012,15(1):34-36.
[23]Coffey J C,Wang J H,Smith M J,et al.Phosphoinositide3-kinase accelerates postoperative tumor growth by inhibiting apop-tosis and enhancing resistance to chemotherapy-induced apoptosisnovel role for an old enemy[J].J Biol Chem,2005,280(22):20968-20977.
[24]Son G,Hines I N,Lindquist J,et al.Inhibition of phosphatidylinositol 3-kinase signaling in hepatic stellate cells blocks the progression of hepatic fibrosis[J].Hepatology,2009,50(5):1512-1523.
[25]Urtasun R,Lopategi A,George J,et al.Osteopontin,an oxidant stress sensitive cytokine,up-regulates collagen-Ivia integrinα(V)β(3)engagement and PI3K/pAkt/NFκBsignaling[J].Hepatology,2012,55(2):594-608.
[26]Peng R,Wang S,Wang R,et al.Antifibrotic effects of tanshinol in experimental hepatic fibrosis by targeting PI3K/AKT/m TOR/p70S6K1 signaling pathways[J].Discov Med,2017,23(125):81-94.
[2]Zhao B.Regulation of TGF-βsignal transduction[J].Scientifica,2014,doi:10.1155/2014/874065.
[3]Kulkarni A A,Thatcher T H,Olsen K C,et al.PPAR-γligands repress TGF-β-induced myofibroblast differentiation by targeting the PI3K/Akt pathway:Implications for therapy of fibrosis[J].PLoS One,2011,6(1):e15909.
[4]Okamoto K,Tajima H,Ohta T,et al.Angiotensin IIinduces tumor progression and fibrosis in intrahepatic cholangiocarcinoma through an interaction with hepatic stellate cells[J].Int J Oncol,2010,37(5):1251-1259.
[5]Mehal W,Imaeda A.Cell death and fibrogenesis[J].Semin Liver Dis,2010,doi:10.1055/s-0030-1255352.
[6]《全国中草药汇编》编写组.全国中草药汇编[M].北京:人民卫生出版社,1978.
[7]Babu B,Shylesh B,Padikkala J.Antioxidant and hepatoprotective effect of Acanthus ilicifolius[J].Fitoterapia,2001,72(3):272-277.
[8]Kumar K M S,Puia Z,Samanta S K,et al.The gastroprotective role of acanthus ilicifolius-a study to unravel the underlying mechanism of anti-ulcer activity[J].Sci Pharm,2012,doi:10.3797/scipharm.1108-11.
[9]Bose S,Bose A.Antimicrobial activity of Acanthus ilicifolius(L.)[J].Ind J Pharm Sci,2008,70(6):821-823.
[10]Babu B,Shylesh B,Padikkala J.Tumour reducing and anticarcinogenic activity of Acanthus ilicifolius in mice[J].J Ethnopharmacol,2002,79(1):27-33.
[11]Sasore T,Reynolds A L,Kennedy B N.Targeting the PI3K/Akt/mTOR Pathway in ocular neovascularization[J].Adv Exp Med Biol,2014,doi:10.1007/978-1-4614-3209-8_101.
[12]Magi S,Saeki Y,Kasamatsu M,et al.Chemical genomic-based pathway analyses for epidermal growth factor-mediated signaling in migrating cancer cells[J].PLoS One,2014,9(5):e96776.
[13]Jung K H,Yan H H,Fang Z,et al.HS-104,a PI3Kinhibitor,enhances the anticancer efficacy of gemcitabine in pancreatic cancer[J].Int J Oncol,2014,45(1):311-321.
[14]Martelli A M,Lonetti A,Buontempo F,et al.Targeting signaling pathways in T-cell acute lymphoblastic leukemia initiating cells[J].Adv Biol Regul,2014,doi:10.1016/j.jbior.2014.04.004.
[15]Engelman J A.Targeting PI3K signaling in cancer:Opportunities,challenges and limitations[J].Nat Rev Cancer,2009,9(8):550-562.
[16]Hao L S,Zhang X L,Zhou Z H,et al.Relationship of PTEN expression withapoptosis of hepatic stellate cells in liver tissues of rats with hepatic fibrosis induced by bile stagnation[J].Med J Chin Lib Army,2010,35(7):836-838.
[17]Yang L,Zhao Z X,Hou J L,et al.Hepatic expression of CB1 in rats with fibrosis and the relationship with FAK[J].J Clin Hepatol,2011,27(8):824-826.
[18]Piguet A C,Dufour J F.PI3K/PTEN/Akt pathway[J].JHepatol,2011,54(6):1317-1319.
[19]G?bele E,Reif S,Tsukada S,et al.The role of p70S6Kinhepatic stellate cell collagen gene expression and cell proliferation[J].J Biol Chem,2005,280(14):13374-13382.
[20]Wei L,Chen Q,Guo A,et al.Asiatic acid attenuates CCl-induced liver fibrosis in rats by regulating the PI3K/AKT/m TOR and Bcl-2/Bax signaling pathways[J].Inter Immunopharmacol,2018,doi:10.1016/j.intimp.2018.04.016.
[21]占书箱,黄成,马陶陶,等.抑制p70S6K对HSC-T6增殖活化的作用研究[J].安徽医科大学学报,2014,49(8):1062-1067.
[22]张坤,姜妙娜,张彩华,等.中药肝复康对肝星状细胞m TOR/P70S6K信号通路的干预作用[J].实用肝脏病杂志,2012,15(1):34-36.
[23]Coffey J C,Wang J H,Smith M J,et al.Phosphoinositide3-kinase accelerates postoperative tumor growth by inhibiting apop-tosis and enhancing resistance to chemotherapy-induced apoptosisnovel role for an old enemy[J].J Biol Chem,2005,280(22):20968-20977.
[24]Son G,Hines I N,Lindquist J,et al.Inhibition of phosphatidylinositol 3-kinase signaling in hepatic stellate cells blocks the progression of hepatic fibrosis[J].Hepatology,2009,50(5):1512-1523.
[25]Urtasun R,Lopategi A,George J,et al.Osteopontin,an oxidant stress sensitive cytokine,up-regulates collagen-Ivia integrinα(V)β(3)engagement and PI3K/pAkt/NFκBsignaling[J].Hepatology,2012,55(2):594-608.
[26]Peng R,Wang S,Wang R,et al.Antifibrotic effects of tanshinol in experimental hepatic fibrosis by targeting PI3K/AKT/m TOR/p70S6K1 signaling pathways[J].Discov Med,2017,23(125):81-94.
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