华蟾素注射液中酯蟾毒配基的分离及体内外抗肿瘤活性筛选
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摘要
目的:通过观察华蟾素注射液中酯蟾毒配基体内外抗肿瘤的作用效果和对部分酯蟾毒配基类单体体外抗胰腺癌细胞的药效筛选,以明确酯蟾毒配基抗胰腺癌的物质基础。方法:运用现代色谱技术分离华蟾素注射液,得到酯蟾毒配基类成分,采用HPLC-DAD-FT-ICR-MS分析并鉴定酯蟾毒配基类化合物的主要单体。通过体外细胞筛选酯蟾毒配基抗肿瘤的活性,在体内建立胰腺癌荷瘤裸鼠模型以评价酯蟾毒配基药效,在体外将分离所得酯蟾毒配基的主要活性单体进行抗胰腺癌活性筛选。结果:华蟾素注射液中的酯蟾毒配基在体外对肝癌BEL7402细胞,胃癌BGC823细胞和胰腺癌SW1990,MIAPaCa-2细胞的半数抑制浓度(IC_(50))分别为(0.47±0.03),(0.06±0.01),(0.06±0.02),(0.11±0.03)g·L~(-1),酯蟾毒配基的半数致死量(LD_(50))为20 mg·kg~(-1),体内对SW1990荷瘤裸鼠的治疗效果接近化疗药物吉西他滨,从酯蟾毒配基中分离并鉴定的12个活性物质体外抗胰腺癌的活性筛选出日蟾毒它灵、沙蟾毒精和嚏根草苷元对SW1990和Bx PC-3细胞具有明显抑制作用。结论:华蟾素注射液中酯蟾毒配基在体外对胰腺癌细胞的抑制率与胃癌几乎相同,在体内对胰腺癌荷瘤裸鼠有明显治疗效果,同时酯蟾毒配基中的单体成分日蟾毒它灵、沙蟾毒精和嚏根草苷元在体外对胰腺癌细胞有明显的抑制作用。
Objective:According to observation of the anti-cancer effect of bufadienolides from cinobufacin injection in vivo and in vitro and the anti-pancreatic cancer cells of bufadienolide monomers in vitro,to indentify the foundation of bufadienolides on anti-pancreatic cancer.Method:Modern chromatographic separation technology was adopted to obtain bufadienolides from cinobufacin injection,HPLC-DAD-FT-ICR-MS was employed to analysis and identify the main monomers of bufadienolides.Anti-cancer activity of bufadienolides was screened by in vitro cells,then setting up the pancreatic cancer animal model in nude mice in vivo for evaluating the efficacy of bufadienolides,the anti-pancreatic cancer activity of the main bufadienolide monomers in vitro was screened.Result:IC_(50) of bufadienolides from cinobufacin injection on hepatocellular carcinoma cells(BEL7402),gastric carcinoma cells(BGC823) and pancreatic cancer cells(SW1990 and MIAPaCa-2) in vitro were(0.47±0.03),(0.06±0.01),(0.06±0.02),(0.11±0.03) g·L~(-1),respectively.LD_(50) of bufadienolides was 20 mg·kg~(-1) and the treatment effect on SW1990 nude mice in vivo was close to gemcitabine.Gamabufotalin,arenobufagin and hellebrigenin had obvious inhibitory effect on SW1990 and BxPC-3 cells,which were isolated and identified from bufadienolides.Conclusion:Bufadienolides from cinobufacin injection shows almost the same inhibition rate on pancreatic cancer cells as gastric cancer in vitro,and it has apparently treatment effect on human pancreatic cancer bearing xenograft nude mice in vivo.Gamabufotalin,arenobufagin and hellebrigenin from bufadienolides show significantly inhibition on pancreatic cancer cells in vitro.
Objective:According to observation of the anti-cancer effect of bufadienolides from cinobufacin injection in vivo and in vitro and the anti-pancreatic cancer cells of bufadienolide monomers in vitro,to indentify the foundation of bufadienolides on anti-pancreatic cancer.Method:Modern chromatographic separation technology was adopted to obtain bufadienolides from cinobufacin injection,HPLC-DAD-FT-ICR-MS was employed to analysis and identify the main monomers of bufadienolides.Anti-cancer activity of bufadienolides was screened by in vitro cells,then setting up the pancreatic cancer animal model in nude mice in vivo for evaluating the efficacy of bufadienolides,the anti-pancreatic cancer activity of the main bufadienolide monomers in vitro was screened.Result:IC_(50) of bufadienolides from cinobufacin injection on hepatocellular carcinoma cells(BEL7402),gastric carcinoma cells(BGC823) and pancreatic cancer cells(SW1990 and MIAPaCa-2) in vitro were(0.47±0.03),(0.06±0.01),(0.06±0.02),(0.11±0.03) g·L~(-1),respectively.LD_(50) of bufadienolides was 20 mg·kg~(-1) and the treatment effect on SW1990 nude mice in vivo was close to gemcitabine.Gamabufotalin,arenobufagin and hellebrigenin had obvious inhibitory effect on SW1990 and BxPC-3 cells,which were isolated and identified from bufadienolides.Conclusion:Bufadienolides from cinobufacin injection shows almost the same inhibition rate on pancreatic cancer cells as gastric cancer in vitro,and it has apparently treatment effect on human pancreatic cancer bearing xenograft nude mice in vivo.Gamabufotalin,arenobufagin and hellebrigenin from bufadienolides show significantly inhibition on pancreatic cancer cells in vitro.
引文
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[13]王丽娟,路军章.中医药治疗胰腺癌的研究进展[J].中华中医药杂志,2016,35(3):961-964.
[14]MENG Z,YANG P,SHEN Y,et al.Pilot study of huachansu in patients with hepatocellular carcinoma,nonsmall-cell lung cancer,or pancreatic cancer[J].Cancer,2011,115(22):5309-5318.
[15]WEI X L,SI N,ZHANG Y,et al.Evaluation of bufadienolides as the main antitumor components in cinobufacin injection for liver and gastric cancer therapy[J].PLo S One,2017,12(1).
[16]魏晓露,杨健,司南,等.华蟾素蟾毒配基类有效组分体内免疫效果分析[J].中国实验方剂学杂志,2016,22(21):87-92.
[2]仲丽丽,白云,费洪新,等.蝙蝠葛酚性碱与胰腺癌研究[J].辽宁中医药大学学报,2014,16(6):85-88.
[3]GUO X Z,CUI Z M,LIU X.Current developments,problems and solutions in the non-surgical treatment of pancreatic cancer[J].World J Gastrointest Oncol,2013,5(2):20-28.
[4]杨尹默.胰腺癌外科治疗的现状、存在问题与展望[J].中国普通外科杂志,2016,25(9):1231-1235.
[5]Blaauw R.Pancreas cancer with Whipple’s operation[J].SAJCN,2015,28(2):92-96.
[6]Jemal A,Bray F,Center M M,et al.Global cancer statistics[J].CA-Cancer J Clin,2011,61(2):69.
[7]邵永孚,赵东兵.胰腺癌诊断治疗中的几个热点问题浅谈[J].中华肝胆外科杂志,2012,18(6):401-403.
[8]Heinemann V,Haas M,Boeck S.Systemic treatment of advanced pancreatic cancer[J].Cancer Treat Rev,2012,38(7):843-853.
[9]Olive K P,Jacobetz M A,Davidson C J,et al.Inhibition of hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer[J].Science,2009,324(5933):1457-1461.
[10]Mimeault M,Batra S K.New advances on critical implications of tumor-and metastasis-initiating cells in cancer progression,treatment resistance and disease recurrence[J].Histol Histopathol,2010,25(8):1057-1073.
[11]杨立新,赵海誉,袁圣芬,等.华蟾素注射液中总蟾毒配基类成分的含量测定[J].中国实验方剂学杂志,2013,19(6):87-89.
[12]郭敬新,李广永,张栋亭,等.中医治疗胰腺癌疗效评价[J].内蒙古中医药,2015(2):12-13.
[13]王丽娟,路军章.中医药治疗胰腺癌的研究进展[J].中华中医药杂志,2016,35(3):961-964.
[14]MENG Z,YANG P,SHEN Y,et al.Pilot study of huachansu in patients with hepatocellular carcinoma,nonsmall-cell lung cancer,or pancreatic cancer[J].Cancer,2011,115(22):5309-5318.
[15]WEI X L,SI N,ZHANG Y,et al.Evaluation of bufadienolides as the main antitumor components in cinobufacin injection for liver and gastric cancer therapy[J].PLo S One,2017,12(1).
[16]魏晓露,杨健,司南,等.华蟾素蟾毒配基类有效组分体内免疫效果分析[J].中国实验方剂学杂志,2016,22(21):87-92.