石柑子水提物对尖吻蝮蛇毒活性抑制作用的初步研究
|
摘要
【目的】寻找用于治疗尖吻蝮(Agkistrodon acutus)咬伤的中草药石柑子(Pothos chinensis)的药效实验证据。【方法】将制备所得石柑子水提物与尖吻蝮蛇毒混合孵育40min,检测混合液中蛇毒的磷脂酶A2活性、透明质酸酶活性、蛋白水解酶活性、出血活性、凝血活性和致死活性。【结果】石柑子水提物对尖吻蝮蛇毒的磷脂酶A2活性、透明质酸酶活性、蛋白水解酶活性、出血活性、凝血活性和致死活性都有明显抑制作用。【结论】研究结果初步证实了石柑子对尖吻蝮咬伤具有治疗效果。
[Purposes]To look for the experimental evidences for the efficacy of P.chinensis,a Chinese herbal medicine,used to treat the envenomings snake bitten by A.acutus.[Methods]After the aqueous extracts were prepared from P.chinensis,and they were incubated with A.acutus venom for 40 min,the activities of PLA2,hyaluronidase,protease and cagulationin the snake venom of the mixtures were examined in vitro assays,as well as hemorrhage activity and lethality activity in the snake venom of the mixtures were detected in vivo assays.[Findings]The aqueous extract of P.chinensis had obvious inhibition against the activities of PLA2,hyaluronidase,protease,hemorrhagic,coagulation and lethality of A.acutus venom.[Conclusions]Preliminary results revealed that P.chinensis has some treatment effects on A.acutus snakebites.
[Purposes]To look for the experimental evidences for the efficacy of P.chinensis,a Chinese herbal medicine,used to treat the envenomings snake bitten by A.acutus.[Methods]After the aqueous extracts were prepared from P.chinensis,and they were incubated with A.acutus venom for 40 min,the activities of PLA2,hyaluronidase,protease and cagulationin the snake venom of the mixtures were examined in vitro assays,as well as hemorrhage activity and lethality activity in the snake venom of the mixtures were detected in vivo assays.[Findings]The aqueous extract of P.chinensis had obvious inhibition against the activities of PLA2,hyaluronidase,protease,hemorrhagic,coagulation and lethality of A.acutus venom.[Conclusions]Preliminary results revealed that P.chinensis has some treatment effects on A.acutus snakebites.
引文
[1]WARRELL D A.Snake bite[J].Lancet,2010,375(9708):77-88.
[2]SANTHOSH M.S,HEMSHEKHAR M,SUNITHA K,et al.Snake venom induced local toxicities:plant secondary metabolites as an auxiliary therapy[J].Mini-Reviews in Medicinal Chemistry,2013,13(1):106-123.
[3]GUTIRREZ J M,LEN G,LOMONTE B,et al.Antivenoms for snakebite envenomings[J].Inflammation&Allergy Drug Targets,2011,10(5):369-380.
[4]GIRISH K S,KEMPARAJU K.Overlooked issues of snakebite management:time for strategic approach[J].Current Topics in Medicinal Chemistry,2011,11(20):2494-2508.
[5]GOMES A,DASR,SARKHEL S,et al.Herbs and herbal constituents active against snake bite[J].Indian Journal of Experimental Biology,2010,48(9):865-78.
[6]MAKHIJA I K,KHAMAR D.Anti-snake venom properties of medicinal plants[J].Der Pharmacia Lettre,2010,2(5):399-411.
[7]GOSWAMI P K,SAMANT M,SRIVASTAVA R S.Snake venom,anti-snake venom&potential of snake venom[J].International Journal of Pharmacy and Pharmaceutical Sciences,2014,6(5):4-7.
[8]SAMY R P,THWIN M M,GOPALAKRISHNAKONE P.Ethnobotanical survey of folk plants for the treatment of snakebites in southern part of Tamilnadu,India[J].Journal of Ethnopharmacology,2008,115(2):302-312.
[9]AHMAD A,ALKHARFY K M,WANI T A,et al.Application of Box-Behnken design for ultrasonic-assisted extraction of polysaccharides from Paeonia emodi[J].International Journal of Biological Macromolecules,2015,72:990-997.
[10]GONZLEZ-CENTENO M R,KNOERZER K,SABAREZ H,et al.Effect of acoustic frequency and power density on the aqueous ultrasonic-assisted extraction of grape pomace(Vitis vinifera L.):a response surface approach[J].Ultrasonics Sonochemistry,2014,21(6):2176.
[11]ROJAS G,GUTIRREZ J M,GENJ A,et al.Neutralization of toxic and enzyme activities of 4venoms from snakes of Guatemala and Honduras by the polyvalent antivenin produced in Costa Rica[J].Revista De Biologia Tropical,1987,35(1):59-67.
[12]FERRANTE N D I.Turbidimetric measurement of acid mucopolysaccharides and hyaluronidase activity[J].Journal of Biological Chemistry,1956,220(1):303-306.
[13]THEAKSTON R D,REID H A.Development of simple standard assay procedures for the characterization of snake venom[J].Bulletin of the World Health Organization,1983,61(6):949-956.
[14]LAING G D,THEAKSTON G,LEITE R P,et al.Comparison of the potency of three Brazilian Bothrops antivenoms using in vivo rodent and in vitro assays.BIASG(Butantan Institute Antivenom Study Group)[J].Toxicon Official Journal of the International Society on Toxinology,1992,30(10):1219-1225.
[15]RODRIGUES V M,SOARES A M,ANDRIOESCARSOS H,et al.Pathological alterations induced by neuwiedase,a metalloproteinase isolated from Bothrops neuwiedi snake venom[J].Biochimie,2001,83(6):471-479.
[16]PAUL K,MANJULA J,DEEPA E P,et al.Anti-Echis carinatus venom antibodies from chicken egg yolk:isolation,purification and neutralization efficacy[J].Toxicon,2007,50(7):893-900.
[17]LISTED N.Progress in the characterization of venoms and standardization of antivenoms[J].Who Offset Publication,1981,58(58):1-44.
[18]KINI R M.Excitement ahead:structure,function and mechanism of snake venom phospholipase A2enzymes[J].Toxicon,2003,42(8):827-840.
[19]MATSUI T,FUJIMURA Y,TITANI K.Snake venom proteases affecting hemostasis and thrombosis[J].Biochimica et Biophysica Acta,2000,1477(1/2):146-156.
[20]PORE S M,RAMANAND S J,PATIL P T,et al.A retrospective study of use of polyvalent anti-snake venom and risk factors for mortality from snake bite in a tertiary care setting[J].Indian Journal of Pharmacology,2015,47(3):270.
[21]GOMES A,SAHA A,CHATTERJEE I,et al.Viper and cobra venom neutralization by beta-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less.(Asteraceae)[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,2007,14(9):637-643.
[22]BORGES M H,SOARES A M,RODRIGUES V M,et al.Effects of aqueous extract of Casearia sylvestris(Flacourtiaceae)on actions of snake and bee venoms and on activity of phospholipases A2[J].Comparative Biochemistry&Physiology B Comparative Biochemistry,2000,127(1):21-30.
[23]BORGES M H,SOARES A M,RODRIGUES V M,et al.Neutralization of proteases from Bothrops,snake venoms by the aqueous extract fromCasearia sylvestris(Flacourtiaceae)[J].Toxicon,2001,39(12):1863-1869.
[24]BIONDO R,PEREIRA A M S,MARCUSSI S,et al.Inhibition of enzymatic and pharmacological activities of some snake venoms and toxins by Mandevilla velutina(Apocynaceae)aqueous extract[J].Biochimie,2003,85(10):1017-1025.
[25]IZIDORO L F M,RODRIGUES V M,RODRIGUES R S,et al.Neutralization of some hematological and hemostatic alterations induced by neuwiedase,a metalloproteinase isolated from Bothrops neuwiedi pauloensis,snake venom,by the aqueous extract from Casearia mariquitensis(Flacourtiaceae)[J].Biochimie,2003,85(7):669-675.
[26]MAIORANO V A,MARCUSSI S,DAHER M A,et al.Antiophidian properties of the aqueous extract of Mikania glomerata[J].Journal of Ethnopharmacology,2005,102(3):364-370.
[27]DA S J,COPPEDE J S,FERNANDES V C,et al.Antihemorrhagic,antinucleolytic and other antiophidian propertiesof the aqueous extract fromPentaclethra macroloba[J].Journal of Ethnopharmacology,2005,100(1/2):145-152.
[28]OLIVEIRA C Z,MAIORANO V A,MARCUSSI S,et al.Anticoagulant and antifibrinogenolytic properties of the aqueous extract from Bauhinia forficata against snake venoms[J].Journal of Ethnopharmacology,2005,98(1/2):213-216.
[29]USHANANDINI S,NAGARAJU S,HARISH K K,et al.The anti-snake venom properties of Tamarindus indica(Leguminosae)seed extract[J].Phytotherapy Research:PTR,2006,20(10):851-858.
[30]GIRISH K S,SHASHIDHARAMURTHY R S,NAGARAJU S,et al.Isolation and characterization of hyaluronidase a"spreading factor"from Indian cobra(Naja naja)venom[J].Biochimie,2004,86(3):193-202.
[31]TU A T,HENDON R R.Characterization of lizard venom hyaluronidase and evidence for its action as a spreading factor[J].Comparative Biochemistry&Physiology B Comparative Biochemistry,1983,76(2):377-383.
[32]GIRISH K S,KEMPARAJU K.Inhibition of Naja naja venom hyaluronidase by plant-derived bioactive components and polysaccharides[J].Biochemistry Biokhimiia,2005,70(8):948-952.
[33]PESSINI A C,TAKAO T T,CAVALHEIRO E C,et al.A hyaluronidase from Tityus serrulatus scorpion venom:isolation,characterization and inhibition by flavonoids[J].Toxicon Official Journal of the International Society on Toxinology,2001,39(10):1495-1504.
[2]SANTHOSH M.S,HEMSHEKHAR M,SUNITHA K,et al.Snake venom induced local toxicities:plant secondary metabolites as an auxiliary therapy[J].Mini-Reviews in Medicinal Chemistry,2013,13(1):106-123.
[3]GUTIRREZ J M,LEN G,LOMONTE B,et al.Antivenoms for snakebite envenomings[J].Inflammation&Allergy Drug Targets,2011,10(5):369-380.
[4]GIRISH K S,KEMPARAJU K.Overlooked issues of snakebite management:time for strategic approach[J].Current Topics in Medicinal Chemistry,2011,11(20):2494-2508.
[5]GOMES A,DASR,SARKHEL S,et al.Herbs and herbal constituents active against snake bite[J].Indian Journal of Experimental Biology,2010,48(9):865-78.
[6]MAKHIJA I K,KHAMAR D.Anti-snake venom properties of medicinal plants[J].Der Pharmacia Lettre,2010,2(5):399-411.
[7]GOSWAMI P K,SAMANT M,SRIVASTAVA R S.Snake venom,anti-snake venom&potential of snake venom[J].International Journal of Pharmacy and Pharmaceutical Sciences,2014,6(5):4-7.
[8]SAMY R P,THWIN M M,GOPALAKRISHNAKONE P.Ethnobotanical survey of folk plants for the treatment of snakebites in southern part of Tamilnadu,India[J].Journal of Ethnopharmacology,2008,115(2):302-312.
[9]AHMAD A,ALKHARFY K M,WANI T A,et al.Application of Box-Behnken design for ultrasonic-assisted extraction of polysaccharides from Paeonia emodi[J].International Journal of Biological Macromolecules,2015,72:990-997.
[10]GONZLEZ-CENTENO M R,KNOERZER K,SABAREZ H,et al.Effect of acoustic frequency and power density on the aqueous ultrasonic-assisted extraction of grape pomace(Vitis vinifera L.):a response surface approach[J].Ultrasonics Sonochemistry,2014,21(6):2176.
[11]ROJAS G,GUTIRREZ J M,GENJ A,et al.Neutralization of toxic and enzyme activities of 4venoms from snakes of Guatemala and Honduras by the polyvalent antivenin produced in Costa Rica[J].Revista De Biologia Tropical,1987,35(1):59-67.
[12]FERRANTE N D I.Turbidimetric measurement of acid mucopolysaccharides and hyaluronidase activity[J].Journal of Biological Chemistry,1956,220(1):303-306.
[13]THEAKSTON R D,REID H A.Development of simple standard assay procedures for the characterization of snake venom[J].Bulletin of the World Health Organization,1983,61(6):949-956.
[14]LAING G D,THEAKSTON G,LEITE R P,et al.Comparison of the potency of three Brazilian Bothrops antivenoms using in vivo rodent and in vitro assays.BIASG(Butantan Institute Antivenom Study Group)[J].Toxicon Official Journal of the International Society on Toxinology,1992,30(10):1219-1225.
[15]RODRIGUES V M,SOARES A M,ANDRIOESCARSOS H,et al.Pathological alterations induced by neuwiedase,a metalloproteinase isolated from Bothrops neuwiedi snake venom[J].Biochimie,2001,83(6):471-479.
[16]PAUL K,MANJULA J,DEEPA E P,et al.Anti-Echis carinatus venom antibodies from chicken egg yolk:isolation,purification and neutralization efficacy[J].Toxicon,2007,50(7):893-900.
[17]LISTED N.Progress in the characterization of venoms and standardization of antivenoms[J].Who Offset Publication,1981,58(58):1-44.
[18]KINI R M.Excitement ahead:structure,function and mechanism of snake venom phospholipase A2enzymes[J].Toxicon,2003,42(8):827-840.
[19]MATSUI T,FUJIMURA Y,TITANI K.Snake venom proteases affecting hemostasis and thrombosis[J].Biochimica et Biophysica Acta,2000,1477(1/2):146-156.
[20]PORE S M,RAMANAND S J,PATIL P T,et al.A retrospective study of use of polyvalent anti-snake venom and risk factors for mortality from snake bite in a tertiary care setting[J].Indian Journal of Pharmacology,2015,47(3):270.
[21]GOMES A,SAHA A,CHATTERJEE I,et al.Viper and cobra venom neutralization by beta-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less.(Asteraceae)[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,2007,14(9):637-643.
[22]BORGES M H,SOARES A M,RODRIGUES V M,et al.Effects of aqueous extract of Casearia sylvestris(Flacourtiaceae)on actions of snake and bee venoms and on activity of phospholipases A2[J].Comparative Biochemistry&Physiology B Comparative Biochemistry,2000,127(1):21-30.
[23]BORGES M H,SOARES A M,RODRIGUES V M,et al.Neutralization of proteases from Bothrops,snake venoms by the aqueous extract fromCasearia sylvestris(Flacourtiaceae)[J].Toxicon,2001,39(12):1863-1869.
[24]BIONDO R,PEREIRA A M S,MARCUSSI S,et al.Inhibition of enzymatic and pharmacological activities of some snake venoms and toxins by Mandevilla velutina(Apocynaceae)aqueous extract[J].Biochimie,2003,85(10):1017-1025.
[25]IZIDORO L F M,RODRIGUES V M,RODRIGUES R S,et al.Neutralization of some hematological and hemostatic alterations induced by neuwiedase,a metalloproteinase isolated from Bothrops neuwiedi pauloensis,snake venom,by the aqueous extract from Casearia mariquitensis(Flacourtiaceae)[J].Biochimie,2003,85(7):669-675.
[26]MAIORANO V A,MARCUSSI S,DAHER M A,et al.Antiophidian properties of the aqueous extract of Mikania glomerata[J].Journal of Ethnopharmacology,2005,102(3):364-370.
[27]DA S J,COPPEDE J S,FERNANDES V C,et al.Antihemorrhagic,antinucleolytic and other antiophidian propertiesof the aqueous extract fromPentaclethra macroloba[J].Journal of Ethnopharmacology,2005,100(1/2):145-152.
[28]OLIVEIRA C Z,MAIORANO V A,MARCUSSI S,et al.Anticoagulant and antifibrinogenolytic properties of the aqueous extract from Bauhinia forficata against snake venoms[J].Journal of Ethnopharmacology,2005,98(1/2):213-216.
[29]USHANANDINI S,NAGARAJU S,HARISH K K,et al.The anti-snake venom properties of Tamarindus indica(Leguminosae)seed extract[J].Phytotherapy Research:PTR,2006,20(10):851-858.
[30]GIRISH K S,SHASHIDHARAMURTHY R S,NAGARAJU S,et al.Isolation and characterization of hyaluronidase a"spreading factor"from Indian cobra(Naja naja)venom[J].Biochimie,2004,86(3):193-202.
[31]TU A T,HENDON R R.Characterization of lizard venom hyaluronidase and evidence for its action as a spreading factor[J].Comparative Biochemistry&Physiology B Comparative Biochemistry,1983,76(2):377-383.
[32]GIRISH K S,KEMPARAJU K.Inhibition of Naja naja venom hyaluronidase by plant-derived bioactive components and polysaccharides[J].Biochemistry Biokhimiia,2005,70(8):948-952.
[33]PESSINI A C,TAKAO T T,CAVALHEIRO E C,et al.A hyaluronidase from Tityus serrulatus scorpion venom:isolation,characterization and inhibition by flavonoids[J].Toxicon Official Journal of the International Society on Toxinology,2001,39(10):1495-1504.