七叶皂苷的制备及其对大鼠后肢水肿的疗效
|
摘要
目的建立高纯度药用级别七叶皂苷的提取工艺并探讨其对组织水肿的治疗作用。方法利用醇提方法从梭罗果中获得七叶皂苷,随后使用软性填料的离子交换柱层析获得高纯度的七叶皂苷单体;使用单体制成喷雾剂对砸伤所致的组织水肿大鼠模型进行治疗。结果从梭罗果中利用醇提获得粗纯的七叶皂苷提取液,活性炭脱色并浓缩后利用离子交换柱层析纯化获得纯度>90%的七叶皂苷单体。以该单体为主要成分制成的喷雾剂对于砸伤所致大鼠组织水肿模型具有良好的治疗效果,显著减轻了后肢的水肿程度(P<0.05),病灶变小,炎性细胞和纤维细胞浸润减少,其效果等同或优于阳性对照药物云南白药。结论传统的用于蛋白质纯化的软琼脂糖凝胶在中药单体的提纯中可发挥重要作用;七叶皂苷钠对于砸伤所致组织水肿有着良好的疗效。
Objective To prepare high purity pharmaceutically acceptable sodium aescinate from Chinese buckeye seed and investigate its therapeutic effect on tissue edema rats. Methods Sodium aescinate was firstly roughly extracted from Chinese buckeye seeds by ethanol and decolorized with activated carbon. Then, the extraction products were further purified by ionexchange chromatography by loading onto a column packed with ion-exchanger medium. The final sodium aescinate products were used as spraying agent to treat tissue edema rat model. Results The purity of sodium aescinate was over 90% after extracted by ethanol and purified via ion-exchange chromatography. When used as spraying agent, sodium aescinate significantly attenuated the degree of edema in the hind limbs of rats, the lesions became smaller, the infiltration of inflammatory cells and fibroblasts was apparently reduced, and the effect was equal or even better than that of the positive control Yunnan Baiyao. Conclusion Soft sepharose beads which are traditionally used for protein purification can be effectively used for Traditional Chinese medicine monomer preparation. Sodium aescinate may be a potential agent for tissue edema treatment.
Objective To prepare high purity pharmaceutically acceptable sodium aescinate from Chinese buckeye seed and investigate its therapeutic effect on tissue edema rats. Methods Sodium aescinate was firstly roughly extracted from Chinese buckeye seeds by ethanol and decolorized with activated carbon. Then, the extraction products were further purified by ionexchange chromatography by loading onto a column packed with ion-exchanger medium. The final sodium aescinate products were used as spraying agent to treat tissue edema rat model. Results The purity of sodium aescinate was over 90% after extracted by ethanol and purified via ion-exchange chromatography. When used as spraying agent, sodium aescinate significantly attenuated the degree of edema in the hind limbs of rats, the lesions became smaller, the infiltration of inflammatory cells and fibroblasts was apparently reduced, and the effect was equal or even better than that of the positive control Yunnan Baiyao. Conclusion Soft sepharose beads which are traditionally used for protein purification can be effectively used for Traditional Chinese medicine monomer preparation. Sodium aescinate may be a potential agent for tissue edema treatment.
引文
[1]Zhou XY, Fu FH, Li Z, et al. Escin, a natural mixture of triterpene saponins,exhibits antitumor activity against hepatocellular carcinoma[J]. Planta Med, 2009, 75(15):1580-5.
[2]Wang YW, Wang SJ, Zhou YN, et al. Escin augments the efficacy of gemcitabine through down-regulation of nuclear factor-κB and nuclear factor-κB-regulated gene products in pancreatic cancer both in vitroandin vivo[J].JCancerResClinOncol,2012,138(5):785-97.
[3]Domanski D, Zegrocka-Stendel O, Perzanowska A, et al. Molecular Mechanism for Cellular Response to β-Escin and Its Therapeutic Implications[J]. PLoS One, 2016, 11(10):e0164365-73.
[4]Chen J, Li W, Yang B, et al. Determination of four major saponins in the seeds of Aesculus chinensis Bunge using accelerated solvent extraction followed by high-performance liquid chromatography and electrospray-time of flight mass spectrometry[J]. Anal Chim Acta, 2007, 596(2):273-80.
[5]Wu X, Liu L, Zhang M, et al. Simultaneous analysis of isomers of escin saponins in human plasma by liquid chromatography-tandem massspectrometry:application to a pharmacokinetic study after oral administration[J].J Chromatogr B Analyt Technol Biomed Life Sci, 2010, 878(11):861-7.
[6]Abudayeh ZH, Al Azzam KM, Naddaf A, et al. Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut(Aesculus Hippocastanum L.)Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography[J]. Adv Pharm Bull, 2015, 5(4):587-91.
[7]Yang XW, Zhao J, Cui YX, et al. Anti-HIV-1 protease triterpenoid saponins from the seeds of Aesculus chinensis[J].J Nat Prod,1999, 62(11):1510-3.
[8]Chen J, Li W, Yang B, et al. Determination of four major saponins in the seeds of Aesculus chinensis Bunge using accelerated solvent extraction followed by high-performance liquid chromatography and electrospray-time of flight mass spectrometry[J]. Anal Chim Acta, 2007, 596(2):273-80.
[9]Li M, Lu C, Zhang L, et al. Oral Administration of Escin Inhibits Acute Inflammation and Reduces Intestinal Mucosal Injury in Animal Models[J].Evid-Based Compl Alte,2015,32(26):503617-25.
[10]Selvakumar GP, Janakiraman U, Essa MM, et al. Escin attenuates behavioral impairments,oxidative stress and inflammation in a chronic MPTP/probenecid mouse model of Parkinson's disease[J].Brain Res, 2014, 1585(1):23-36.
[11]Matsuda H, Li Y, Yoshikawa M. Gastroprotections of escins Ia, Ib,IIa, and IIb on ethanol-induced gastric mucosal lesions in rats[J].Eur J Pharmacol, 1999, 373(1):63-70.
[12]Wang T, Zhao S, Wang Y, et al. Protective effects of escin against indomethacin-induced gastric ulcer in mice[J].ToxicolMech Methods, 2014, 24(8):560-6.
[13]Wang T,Jiang N,Han B,et al.Escin attenuates cerebral edema induced by acute omethoate poisoning[J]. Toxicol Mech Methods,2011, 21(4):400-5.
[14]Yu Z,Su P.Effect of beta-aescin extract from Chinese buckeye seed on chronic venous insufficiency[J].Die Pharmazie,2013,68(3):428-30.
[15]任捷,吴彦,刘海平.七叶皂苷钠片治疗外伤性肢体肿胀的临床观察[J].中国基层医药, 2011, 18(2):38-9.
[16]王梦炎,李会晨,石汉平,等.β-七叶皂甙钠预防术后肠粘连的实验研究[J].中国中西医结合外科杂志, 1999, 6(3):382-4.
[17]杨秀伟,赵静,崔景荣,等.七叶树皂苷-Ia的人肠内细菌生物转化产物及其抗肿瘤活性研究[J].北京大学学报(医学版), 2004,39(1):31-5.
[18]Tan SM, Li F, Rajendran P, et al. Identification of beta-escin as a novel inhibitor of signal transducer and activator of transcription 3/Janus-activated kinase 2 signaling pathway that suppresses proliferation and induces apoptosis in human hepatocellular carcinoma cells[J]. J Pharmacol Exp Ther, 2010, 334(1):285-93.
[19]周庆博,毕建忠,李鲁扬,等.β-七叶皂甙钠对大鼠脑出血后血SOD、 NO、 TNF-α的影响[J].山东大学学报(医学版),2005,43(1):44-7.
[20]Cheng Y, Wang H, Mao M, et al. Escin Increases the Survival Rate of LPS-Induced Septic Mice Through Inhibition of HMGB1 Release from Macrophages[J]. Cell Physiol Biochem, 2015, 36(4):1577-86.
[21]王绪英.娑罗子中七叶皂苷的提取分离与纯化[J].西北植物学报, 2003, 23(9):1592-5.
[22]王绪英,赵永芳.探讨NKA-9树脂分离七叶皂苷的效果[J].武汉植物学研究, 2002, 20(3):225-8.
[23]陈新艳,朱泱平.浙江七叶树种子中七叶皂苷的分离纯化[J].中药新药与临床药理, 2010, 21(3):307-11.
[24]湛建峰,牛剑钊,张启明,等.七叶皂苷A、B的制备液相色谱法分离和结构确证[J].中国药事, 2009, 23(12):1184-6.
[25]Cheng P,Kuang F,Ju G.Aescin reduces oxidative stress and provides neuroprotection in experimental traumatic spinalcord injury[J]. Free Radic Biol Med, 2016, 99(4):405-17.
[26]Asgarirad H,Chabra A,Rahimnejad M,et al.Comparison of albizia julibressin and silver sulfadiazine in healing of second and third degree burns[J]. World J Plas Surg, 2015, 7(1):34-44.
[27]Zhao SQ,Xu SQ,Cheng J,et al.Anti-inflammatory effect of external use of escin on cutaneous inflammation:possible involvement of glucocorticoids receptor[J]. Chin J Nat Med, 2018,16(2):105-12.
[28]Wang T,Jiang N,Han B,et al.Escin attenuates cerebral edema induced by acute omethoate poisoning[J]. Toxicol Mech Methods,2011, 21(5):400-5.
[29]Xiao GM,Wei J.Effects of beta-Aescin on the expression of nuclear factor-kappaB and tumor necrosis factor-alpha after traumatic brain injury in rats[J]. J Zhejiang Univ Sci B, 2005, 6(1):28-32.
[30]Mingrino S, Scanarini M. Anti-edema effect of escin at the cerebral level[J]. Minerva Anestesiol, 1978, 44(6):355-60.
[31]Ren X, Zhang M, Chen L, et al. The anti-inflammatory effects of Yunnan Baiyao are involved inregulation of the phospholipase A2/arachidonic acid metabolites pathways in acute inflammation rat model[J]. Mol Med Rep, 2017, 16(4):4045-53.
[32]Frederick J,Boysen S,Wagg C,et al.The effects of oral administration of Yunnan Baiyao on blood coagulation in beagle dogs as measured by kaolin-activated thromboelastography and buccal mucosal bleeding times[J]. Can J Vet Res, 2017, 81(1):41-5.
[2]Wang YW, Wang SJ, Zhou YN, et al. Escin augments the efficacy of gemcitabine through down-regulation of nuclear factor-κB and nuclear factor-κB-regulated gene products in pancreatic cancer both in vitroandin vivo[J].JCancerResClinOncol,2012,138(5):785-97.
[3]Domanski D, Zegrocka-Stendel O, Perzanowska A, et al. Molecular Mechanism for Cellular Response to β-Escin and Its Therapeutic Implications[J]. PLoS One, 2016, 11(10):e0164365-73.
[4]Chen J, Li W, Yang B, et al. Determination of four major saponins in the seeds of Aesculus chinensis Bunge using accelerated solvent extraction followed by high-performance liquid chromatography and electrospray-time of flight mass spectrometry[J]. Anal Chim Acta, 2007, 596(2):273-80.
[5]Wu X, Liu L, Zhang M, et al. Simultaneous analysis of isomers of escin saponins in human plasma by liquid chromatography-tandem massspectrometry:application to a pharmacokinetic study after oral administration[J].J Chromatogr B Analyt Technol Biomed Life Sci, 2010, 878(11):861-7.
[6]Abudayeh ZH, Al Azzam KM, Naddaf A, et al. Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut(Aesculus Hippocastanum L.)Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography[J]. Adv Pharm Bull, 2015, 5(4):587-91.
[7]Yang XW, Zhao J, Cui YX, et al. Anti-HIV-1 protease triterpenoid saponins from the seeds of Aesculus chinensis[J].J Nat Prod,1999, 62(11):1510-3.
[8]Chen J, Li W, Yang B, et al. Determination of four major saponins in the seeds of Aesculus chinensis Bunge using accelerated solvent extraction followed by high-performance liquid chromatography and electrospray-time of flight mass spectrometry[J]. Anal Chim Acta, 2007, 596(2):273-80.
[9]Li M, Lu C, Zhang L, et al. Oral Administration of Escin Inhibits Acute Inflammation and Reduces Intestinal Mucosal Injury in Animal Models[J].Evid-Based Compl Alte,2015,32(26):503617-25.
[10]Selvakumar GP, Janakiraman U, Essa MM, et al. Escin attenuates behavioral impairments,oxidative stress and inflammation in a chronic MPTP/probenecid mouse model of Parkinson's disease[J].Brain Res, 2014, 1585(1):23-36.
[11]Matsuda H, Li Y, Yoshikawa M. Gastroprotections of escins Ia, Ib,IIa, and IIb on ethanol-induced gastric mucosal lesions in rats[J].Eur J Pharmacol, 1999, 373(1):63-70.
[12]Wang T, Zhao S, Wang Y, et al. Protective effects of escin against indomethacin-induced gastric ulcer in mice[J].ToxicolMech Methods, 2014, 24(8):560-6.
[13]Wang T,Jiang N,Han B,et al.Escin attenuates cerebral edema induced by acute omethoate poisoning[J]. Toxicol Mech Methods,2011, 21(4):400-5.
[14]Yu Z,Su P.Effect of beta-aescin extract from Chinese buckeye seed on chronic venous insufficiency[J].Die Pharmazie,2013,68(3):428-30.
[15]任捷,吴彦,刘海平.七叶皂苷钠片治疗外伤性肢体肿胀的临床观察[J].中国基层医药, 2011, 18(2):38-9.
[16]王梦炎,李会晨,石汉平,等.β-七叶皂甙钠预防术后肠粘连的实验研究[J].中国中西医结合外科杂志, 1999, 6(3):382-4.
[17]杨秀伟,赵静,崔景荣,等.七叶树皂苷-Ia的人肠内细菌生物转化产物及其抗肿瘤活性研究[J].北京大学学报(医学版), 2004,39(1):31-5.
[18]Tan SM, Li F, Rajendran P, et al. Identification of beta-escin as a novel inhibitor of signal transducer and activator of transcription 3/Janus-activated kinase 2 signaling pathway that suppresses proliferation and induces apoptosis in human hepatocellular carcinoma cells[J]. J Pharmacol Exp Ther, 2010, 334(1):285-93.
[19]周庆博,毕建忠,李鲁扬,等.β-七叶皂甙钠对大鼠脑出血后血SOD、 NO、 TNF-α的影响[J].山东大学学报(医学版),2005,43(1):44-7.
[20]Cheng Y, Wang H, Mao M, et al. Escin Increases the Survival Rate of LPS-Induced Septic Mice Through Inhibition of HMGB1 Release from Macrophages[J]. Cell Physiol Biochem, 2015, 36(4):1577-86.
[21]王绪英.娑罗子中七叶皂苷的提取分离与纯化[J].西北植物学报, 2003, 23(9):1592-5.
[22]王绪英,赵永芳.探讨NKA-9树脂分离七叶皂苷的效果[J].武汉植物学研究, 2002, 20(3):225-8.
[23]陈新艳,朱泱平.浙江七叶树种子中七叶皂苷的分离纯化[J].中药新药与临床药理, 2010, 21(3):307-11.
[24]湛建峰,牛剑钊,张启明,等.七叶皂苷A、B的制备液相色谱法分离和结构确证[J].中国药事, 2009, 23(12):1184-6.
[25]Cheng P,Kuang F,Ju G.Aescin reduces oxidative stress and provides neuroprotection in experimental traumatic spinalcord injury[J]. Free Radic Biol Med, 2016, 99(4):405-17.
[26]Asgarirad H,Chabra A,Rahimnejad M,et al.Comparison of albizia julibressin and silver sulfadiazine in healing of second and third degree burns[J]. World J Plas Surg, 2015, 7(1):34-44.
[27]Zhao SQ,Xu SQ,Cheng J,et al.Anti-inflammatory effect of external use of escin on cutaneous inflammation:possible involvement of glucocorticoids receptor[J]. Chin J Nat Med, 2018,16(2):105-12.
[28]Wang T,Jiang N,Han B,et al.Escin attenuates cerebral edema induced by acute omethoate poisoning[J]. Toxicol Mech Methods,2011, 21(5):400-5.
[29]Xiao GM,Wei J.Effects of beta-Aescin on the expression of nuclear factor-kappaB and tumor necrosis factor-alpha after traumatic brain injury in rats[J]. J Zhejiang Univ Sci B, 2005, 6(1):28-32.
[30]Mingrino S, Scanarini M. Anti-edema effect of escin at the cerebral level[J]. Minerva Anestesiol, 1978, 44(6):355-60.
[31]Ren X, Zhang M, Chen L, et al. The anti-inflammatory effects of Yunnan Baiyao are involved inregulation of the phospholipase A2/arachidonic acid metabolites pathways in acute inflammation rat model[J]. Mol Med Rep, 2017, 16(4):4045-53.
[32]Frederick J,Boysen S,Wagg C,et al.The effects of oral administration of Yunnan Baiyao on blood coagulation in beagle dogs as measured by kaolin-activated thromboelastography and buccal mucosal bleeding times[J]. Can J Vet Res, 2017, 81(1):41-5.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |