菊苣酸对蓖麻毒素活性的抑制作用研究
|
摘要
为了解菊苣酸对蓖麻毒素活性的抑制作用,通过硫酸铵沉淀法和亲和层析技术,纯化得到活性良好、蛋白纯度较高的蓖麻毒素活性蛋白。将菊苣酸与蓖麻毒素共孵育,通过LDH检测发现,菊苣酸可以显著抑制蓖麻毒素活性,在药物质量浓度为64 mg·L~(-1)时,LDH释放率为38%。通过动物实验,发现菊苣酸能够降低蓖麻毒素中毒引起的小鼠死亡,死亡率为50%。研究结果表明,菊苣酸通过抑制蓖麻毒素活性,降低细胞LDH释放率,降低小鼠死亡率。表明菊苣酸是一种治疗蓖麻毒素中毒的潜在天然化合物。
The aim of this study is to determine the inhibiting effect of chicoric acid on ricin toxin activity. In this study,sulfate precipitation method and affinity formation technology were used to get highly purified ricin of great bio-activity. Through LDH experiment,we found that the bioactivity of ricin could be inhibited by cichoric acid. Through animal experiment,we evaluated the protective effect of chicoric acid on ricin-poisoned mice. The activity of purified ricin protein is good and of high purity. Through incubation of cichoric acid and ricin,we found cichoric acid can significantly inhibit the activity of ricin,64 mg·L~(-1) cichoric acid induced LDH release 38%. Through animal experiment,we found that cichoric acid could reduce mouse mortality,in which 50% of the animals death. The results of this study indicated that cichoric acid could inhibit the activity of ricin,and reduce the mortality of mice,which indicated that cichoric acid is a kind of potential natural therapeutic compounds of ricin poisoning.
The aim of this study is to determine the inhibiting effect of chicoric acid on ricin toxin activity. In this study,sulfate precipitation method and affinity formation technology were used to get highly purified ricin of great bio-activity. Through LDH experiment,we found that the bioactivity of ricin could be inhibited by cichoric acid. Through animal experiment,we evaluated the protective effect of chicoric acid on ricin-poisoned mice. The activity of purified ricin protein is good and of high purity. Through incubation of cichoric acid and ricin,we found cichoric acid can significantly inhibit the activity of ricin,64 mg·L~(-1) cichoric acid induced LDH release 38%. Through animal experiment,we found that cichoric acid could reduce mouse mortality,in which 50% of the animals death. The results of this study indicated that cichoric acid could inhibit the activity of ricin,and reduce the mortality of mice,which indicated that cichoric acid is a kind of potential natural therapeutic compounds of ricin poisoning.
引文
[1]郎立伟,王玉霞.蓖麻毒素检测方法研究进展[J].军事医学科学院院刊,2009,33(6):573-576.
[2] VITETTA E S,CUSHLEY W,UHR J W. Synergy of ricin A chain-containing immunotoxins and ricin B chaincontaining immunotoxins in in vitro killing of neoplastic human B cells[J]. Proc Natl Acad Sci USA,1983,80(20):6332-6335.
[3] AUDI J,BELSON M,PATEL M,et al. Ricin poisoning:a comprehensive review[J]. The Journal of the American Medical Association,2005,294(18):2342-2351.
[4] ROTZ L D,KHAN A S,LILLIBRIDGE S R,et al. Public health assessment of potential biological terrorism agents[J]. Emerg Infect Dis,2002,8(2):126-132.
[5] RASOOLY R,HE X,FRIEDMAN M. Milk inhibits the biological activity of ricin[J]. J Biol Chem,2012,287(33):27924-27929.
[6] SPITLER L E,RIO M,KHENTIGAN A,et al. Therapy of patients with malignant melanoma using a monoclonal antimelanoma antibody-ricin A chain immunotoxin[J].Cancer Res,1987,47(6):1717-1723.
[7] AMLOT P L,STONE M J,CUNNINGHAM D,et al. A phase I study of an anti-CD22-deglycosylated ricin A chain immunotoxin in the treatment of B-cell lymphomas resistant to conventional therapy[J]. Blood,1993,82(9):2624-2633.
[8] WAHOME P G,ROBERTUS J D,MANTIS N J. Smallmolecule inhibitors of ricin and Shiga toxins[J]. Curr Top Microbiol Immunol,2012,357:179-207.
[9] STECHMANN B,BAI S K,GOBBO E,et al. Inhibition of retrograde transport protects mice from lethal ricin challenge[J]. Cell,2010,141:231-242.
[10] SPOONER R A,LORD J M. How ricin and Shiga toxin reach the cytosol of target cells:retrotranslocation from the endoplasmic reticulum[J]. Curr Top Microbiol Immunol,2013,357:19-40.
[11] DONG J,ZHANG Y,CHEN Y,et al,Baicalin inhibits the lethality of ricin in mice by inducing protein oligomerization[J]. J Biol Chem,2015,290(20):12899-12907.
[12]王立娜.大青木香肾毒性研究及高内涵肾细胞毒性筛选方法的建立[D].北京:北京中医药大学,2016.
[13] LU Q B,WAN M Y,WANG P Y,et al. Chicoric acid prevents PDGF-BB-induced VSMC dedifferentiation,proliferation and migration by suppressing ROS/NFκB/m TOR/P70S6K signaling cascade[J]. Redox Biol,2018,14:656-668.
[14] CHEN L,HUANG G,GAO M,et al. Chicoric acid suppresses BAFF expression in B lymphocytes by inhibiting NF-κB activity[J]. Int Immunopharmacol,2017,44:211-215.
[15] ZOU L B,ZHAN J B. Purification and anti-cancer activity of ricin[J]. Zhejiang Daxue Xuebao(Yixue Ban),2005,34(3):217-219.
[16] GAUCHER S,JARRAYA M. Technical note:comparison of the Presto Blue and LDH release assays with the MTT assay for skin viability assessment[J]. Cell Tissue Bank,2015,16(3):325-329.
[17]高彬.蓖麻毒素的分离提取及活性研究[D].成都:四川大学,2007.
[18]罗嘉惠.蓖麻毒素的研究与应用[J].氨基酸和生物资源,1999,21(2):56-58.
[19] RUTENBER E,READY M,ROBERTUS J D. Structure and evolution of ricin B chain[J]. Nature,1987,326:624-626.
[20] THYGESENJ L,THULIN J,MORTENSEN A,et al. Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea,alone and in combination[J]. Food Chemistry,2007,101(1):74-81.
[2] VITETTA E S,CUSHLEY W,UHR J W. Synergy of ricin A chain-containing immunotoxins and ricin B chaincontaining immunotoxins in in vitro killing of neoplastic human B cells[J]. Proc Natl Acad Sci USA,1983,80(20):6332-6335.
[3] AUDI J,BELSON M,PATEL M,et al. Ricin poisoning:a comprehensive review[J]. The Journal of the American Medical Association,2005,294(18):2342-2351.
[4] ROTZ L D,KHAN A S,LILLIBRIDGE S R,et al. Public health assessment of potential biological terrorism agents[J]. Emerg Infect Dis,2002,8(2):126-132.
[5] RASOOLY R,HE X,FRIEDMAN M. Milk inhibits the biological activity of ricin[J]. J Biol Chem,2012,287(33):27924-27929.
[6] SPITLER L E,RIO M,KHENTIGAN A,et al. Therapy of patients with malignant melanoma using a monoclonal antimelanoma antibody-ricin A chain immunotoxin[J].Cancer Res,1987,47(6):1717-1723.
[7] AMLOT P L,STONE M J,CUNNINGHAM D,et al. A phase I study of an anti-CD22-deglycosylated ricin A chain immunotoxin in the treatment of B-cell lymphomas resistant to conventional therapy[J]. Blood,1993,82(9):2624-2633.
[8] WAHOME P G,ROBERTUS J D,MANTIS N J. Smallmolecule inhibitors of ricin and Shiga toxins[J]. Curr Top Microbiol Immunol,2012,357:179-207.
[9] STECHMANN B,BAI S K,GOBBO E,et al. Inhibition of retrograde transport protects mice from lethal ricin challenge[J]. Cell,2010,141:231-242.
[10] SPOONER R A,LORD J M. How ricin and Shiga toxin reach the cytosol of target cells:retrotranslocation from the endoplasmic reticulum[J]. Curr Top Microbiol Immunol,2013,357:19-40.
[11] DONG J,ZHANG Y,CHEN Y,et al,Baicalin inhibits the lethality of ricin in mice by inducing protein oligomerization[J]. J Biol Chem,2015,290(20):12899-12907.
[12]王立娜.大青木香肾毒性研究及高内涵肾细胞毒性筛选方法的建立[D].北京:北京中医药大学,2016.
[13] LU Q B,WAN M Y,WANG P Y,et al. Chicoric acid prevents PDGF-BB-induced VSMC dedifferentiation,proliferation and migration by suppressing ROS/NFκB/m TOR/P70S6K signaling cascade[J]. Redox Biol,2018,14:656-668.
[14] CHEN L,HUANG G,GAO M,et al. Chicoric acid suppresses BAFF expression in B lymphocytes by inhibiting NF-κB activity[J]. Int Immunopharmacol,2017,44:211-215.
[15] ZOU L B,ZHAN J B. Purification and anti-cancer activity of ricin[J]. Zhejiang Daxue Xuebao(Yixue Ban),2005,34(3):217-219.
[16] GAUCHER S,JARRAYA M. Technical note:comparison of the Presto Blue and LDH release assays with the MTT assay for skin viability assessment[J]. Cell Tissue Bank,2015,16(3):325-329.
[17]高彬.蓖麻毒素的分离提取及活性研究[D].成都:四川大学,2007.
[18]罗嘉惠.蓖麻毒素的研究与应用[J].氨基酸和生物资源,1999,21(2):56-58.
[19] RUTENBER E,READY M,ROBERTUS J D. Structure and evolution of ricin B chain[J]. Nature,1987,326:624-626.
[20] THYGESENJ L,THULIN J,MORTENSEN A,et al. Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea,alone and in combination[J]. Food Chemistry,2007,101(1):74-81.