土贝母苷甲对单纯疱疹病毒1型和2型的增殖和基因组复制的抑制效应
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摘要
土贝母苷甲(Tubeimoside 1,TBMS1),简称苷甲,是一种独特的,由贝咢苷元和4种单糖(葡萄糖、鼠李糖、木糖和阿拉伯糖)组成的五环三萜皂苷。以前的研究证实,苷甲具有抗人免疫缺陷病毒的活性。本文研究苷甲对单纯疱疹病毒(Herpes simplex virus,HSV)1型和2型增殖和基因组复制的影响。从土贝母块茎中提取、分离苷甲(在本实验中使用的纯度大于95%),以空斑法测定病毒滴度,通过观察致细胞病变效应和空班形成抑制试验来检测苷甲的抗病毒活性,用实时定量PCR技术检测苷甲对HSV-1和HSV-2基因组复制的影响。结果显示,苷甲显著抑制HSV-1和HSV-2毒株的增殖,显著抑制HSV-1和HSV-2的UL27、UL52和UL54三个基因的拷贝数,且其抑制作用呈明显量效依赖关系。无毒高浓度(0.39μg/mL～0.78μg/mL)的苷甲对阿昔洛韦(acyclovir,ACV)耐药毒株HSV-1/106的增殖也显示抑制活性。本研究结果揭示苷甲具有显著抑制HSV-1和HSV-2增殖及基因组复制的活性,无毒高浓度苷甲对HSV-1/106耐药毒株的增殖也显示抑制活性。
Tubeimoside(TBMS)1 is a unique pentacyclic triterpenoid saponin composed of bayogenin and four sugars:glucose,rhamnose,xylose and arabinose. Previous studies have shown that TBMS1 has activity against the human immunodeficiency virus. We investigated the inhibitory effects of TBMS1 on the proliferation and genomic replication of herpes simplex virus type 1(HSV-1)and 2(HSV-2). TBMS1 is extracted and isolated from the tubers of[Bolbostemma paniculatum(Maxim)Franquet(Cucurbitaceae)],and its purity in our study was great than 95%. The toxicity of HSV-1 and HSV-2 was determined by the plaque assay. The antiviral activity of TBMS1 was determined by cytopathic effects and plaque-inhibition test,and quantitative real-time polymerase chain reaction was used to detect the effects of TBMS1 on HSV genomic replication.TBMS1 showed significantly inhibitory effects on the proliferation of HSV-1/F and HSV-2/333 strains,and on copies of UL27,UL52 and UL54 in a dose-dependent manner. TBMS1 of high concentration(0.39 μg/mL～0.78 μ g/mL) was not toxic and showed inhibitory effects on the proliferation of the ACV-resistant strain HSV-1/106. TBMS1 had obvious inhibitory effects on the proliferation and genomic replication of HSV-1 and HSV-2.
Tubeimoside(TBMS)1 is a unique pentacyclic triterpenoid saponin composed of bayogenin and four sugars:glucose,rhamnose,xylose and arabinose. Previous studies have shown that TBMS1 has activity against the human immunodeficiency virus. We investigated the inhibitory effects of TBMS1 on the proliferation and genomic replication of herpes simplex virus type 1(HSV-1)and 2(HSV-2). TBMS1 is extracted and isolated from the tubers of[Bolbostemma paniculatum(Maxim)Franquet(Cucurbitaceae)],and its purity in our study was great than 95%. The toxicity of HSV-1 and HSV-2 was determined by the plaque assay. The antiviral activity of TBMS1 was determined by cytopathic effects and plaque-inhibition test,and quantitative real-time polymerase chain reaction was used to detect the effects of TBMS1 on HSV genomic replication.TBMS1 showed significantly inhibitory effects on the proliferation of HSV-1/F and HSV-2/333 strains,and on copies of UL27,UL52 and UL54 in a dose-dependent manner. TBMS1 of high concentration(0.39 μg/mL～0.78 μ g/mL) was not toxic and showed inhibitory effects on the proliferation of the ACV-resistant strain HSV-1/106. TBMS1 had obvious inhibitory effects on the proliferation and genomic replication of HSV-1 and HSV-2.
引文
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[14]Dua Q,Gub Z,Lenevad I,Jianga H,Lia R,Dengb L,Yang Z.The antiviral activity of arbidol hydrochloride against herpes simplex virus type II(HSV-2)in a mouse model of vaginitis[J].Int Immunopharmacol,2019,68:58-67.
[15]Liu T,Gao H,Chen X,Lou G,Gu L,Yang M,Xia B,Yin H.TNFAIP8 as a predictor of metastasis and a novel prognostic biomarker in patients with epithelial ovarian cancer[J].Br J Cancer,2013,109:1685-1692.
[16]Inoue H,Mori T,Shibata S,Koshihara Y.Inhibitory effect of glycyrrhetinic acid derivatives on arachidonic acid-induced mouse ear oedema[J].J Pharm Pharmacol,1988,40(4):272-277.
[17]Inoue H,Mori T,Shibata S,Koshihara Y.Modulation by glycyrrhetinic acid derivatives of TPA-induced mouse ear oedema[J].Br J Pharmacol,1989,96(1):204-210.
[18]于立坚,马润娣,于廷曦.化妆品新原料土贝母总皂苷[P].中国专利,申请号:201510308579.2
[19]于立坚,马润娣,于廷曦.化妆品新原料土贝母总皂苷消除臭汗症臭味的用途[P].中国专利,申请号:201510308559.5
[2]Kong F,Zhu D,Xu R.Structural study of tubeimoside1,a constituent of tu-bei-mu[J].Tetrahedron Lett,1986,27(47):5765-5769.
[3]Yu T X,Ma R D,Yu L J.Structure-activity relationship of tubeimosides in anti-inflammatory,antitumor,and antitumor-promoting effects[J].Acta Pharmacol Sin,2001,22(5):463-468.
[4]Yu L J,Ma R D,Wang Y Q,Nishino H,Takayasu J,He W Z,Chang M,Zhen J,Liu W S,Fan S X.Potent anti-tumorigenic effect of tubeimoside 1 isolated from the bulb of Bolbostemma paniculatum(Maxim)Franquet[J].Itn J Cancer,1992,50(4):635-638.
[5]Yu L J,Ma R D,Wang Y Q,Nishino H.Potent antitumor activity and low toxicity of tubeimoside 1 isolated from Bolbostemma paniculatum[J].Planta Med,1994,60(3):204-208.
[6]于立坚,张永平,马润娣,于廷曦.土贝母皂苷抑制对苯二胺诱发豚鼠接触性皮炎效应的研究[J].中国免疫学杂志,2016,32(2):1626-1631.
[7]Yu L J,Ma R D,Jiang S B.Effects of tubeimoside 1 on HIV core protein P24 and cytopathogenesis in vitro[J].Acta Pharmacol Sin,1994,15:103-106.
[8]左联,谢鹏,徐霞红,胡晖,顾晓波,黄东雅.Balb/c小鼠单纯疱疹病毒性脑炎模型的建立[J].蚌埠医学院学报,2013,38(2):131-133.
[9]孙朝晖,石玉玲,冼江,王捷,杨慧兰.HSV-2/333株在Vero细胞中持续感染及增殖特性研究[J].医学临床研究,2009,26(5):753-755.
[10]Wang Y,Wang Q,Zhu Q,Zhou R,Liu J,Peng T.Identification and characterization of acyclovir-resistant clinical HSV-1 isolates from children[J].J Clin Virol,2011,52:107-112.
[11]韩先杰,王宏伟,王金宝.猪繁殖与呼吸综合征病毒TCID50的测定[J].青岛农业大学学报(自然科学版),2005,22(2):132-134.
[12]殷震,刘景华主编.动物病毒学[M].北京:科学出版社.第2版,1997,329-330.
[13]邹毅,黄海鹭,徐军,钟南山.小鼠的原发性呼吸道合胞病毒感染[J].中华结核和呼吸杂志,2001,24(08):487-489.
[14]Dua Q,Gub Z,Lenevad I,Jianga H,Lia R,Dengb L,Yang Z.The antiviral activity of arbidol hydrochloride against herpes simplex virus type II(HSV-2)in a mouse model of vaginitis[J].Int Immunopharmacol,2019,68:58-67.
[15]Liu T,Gao H,Chen X,Lou G,Gu L,Yang M,Xia B,Yin H.TNFAIP8 as a predictor of metastasis and a novel prognostic biomarker in patients with epithelial ovarian cancer[J].Br J Cancer,2013,109:1685-1692.
[16]Inoue H,Mori T,Shibata S,Koshihara Y.Inhibitory effect of glycyrrhetinic acid derivatives on arachidonic acid-induced mouse ear oedema[J].J Pharm Pharmacol,1988,40(4):272-277.
[17]Inoue H,Mori T,Shibata S,Koshihara Y.Modulation by glycyrrhetinic acid derivatives of TPA-induced mouse ear oedema[J].Br J Pharmacol,1989,96(1):204-210.
[18]于立坚,马润娣,于廷曦.化妆品新原料土贝母总皂苷[P].中国专利,申请号:201510308579.2
[19]于立坚,马润娣,于廷曦.化妆品新原料土贝母总皂苷消除臭汗症臭味的用途[P].中国专利,申请号:201510308559.5