In vitro inhibitory effects of papain on platelet aggregation

Abstract

Abstract: AIM: To observe the in vitro inhibito-ry effects of papain on platelet aggregation by washed platelet model, and explore the possible antithrombotic mechanism of papain .METHODS: Human washed platelets were mixed with different doses of papain. The maximal ratio of platelet aggregation induced by 5-adenosine liphosphate-disodium salt (ADP) , arachi-donic acid (AA) , collagen and thrombin, respective-ly, was measured with platelet aggregation analyzer. The expressions of fibrinogen receptor(FIB-R) and p-selectin(CD62p) on external membrane of activated platelet were determinated with flow cytometry .The changes of factin cytoskeletal structure in activated platelet were analyzed by SDS-PAGE .Platelet rich plasma(PRP) of patients with primary hypertension (PH) or acute myocardiac infarction(AMI) was treated to determinate the levels of aggregation and FIB-R ex-pression.RESULTS: Papain significantly inhibited platelet aggregation in a dose-dependent manner and reduced the levels of platelet aggregation, there was re-markably negative correlation between platelet aggrega-tion level and papain dose(P<0.01) .Papain notably inhibited FIB-R expression and decreased the levels of FIB-R expression activated by ADP in a dose-depen-dent manner.Papain reduced P-selectin expression on external membrane and lessened f-actins of platelet triggered by ADP .Papain significantly inhibited the levels of aggregation and FIB-R expression of platelets activated by ADP in patients with PH and AMI(P<0.01) .CONCLUSION: Papain can inhibit the ex-pression of fibrinogen receptor on external membrane of activated platelet and polymerization or release reac-tion, so the thrombocytic aggregation are inhibited in a dose-dependent manner, and papain has the effect of anti-thrombogenesis.

Key words: papain, platelet aggregation, fibrinogen receptor, P-selectin, actin polymer

CLC Number:

R965.2

FEI Xian-ming, ZHOU Yong-lie, QI Jin-wen, WU Jian-guo, QIU Lian-nv, WANG Zhen-ni. In vitro inhibitory effects of papain on platelet aggregation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(8): 906-911.

References

[1] 张丽娜, 凌沛学, 娄红祥, 等.木瓜酶及其在医药中的应用[J] .药学实践, 2007, 25(6):361-363.
[2] 张勇, 石荣书, 钱民章.木瓜蛋白酶体内溶栓效果观察[J] .遵义医学院学报, 2007, 30(s1):1-3.
[3] 吴源, 郭宏宝, 王铁军, 等.几种中药组分对家兔体外血小板聚集作用的比较[J] .中国临床药理学与治疗学, 2007, 12(9):1047-1050.
[4] Giovanni D, Carlo P.Platelet activation and atherothrom-bosis[J] .N Engl J Med, 2007, 357(24):2482-2494.
[5] Offermanns S.Activation of platelet function through G protein-coupled receptors[J] .Circ Res, 2006, 99(12): 1293-1304.
[6] RandML, Leung R, Packham MA, et al.Platelet func-tion assays[J] .Transfus Apher Sci, 2003, 28(3):307-317.
[7] Nieswandt B, Watson SP.Platelet-collagen interaction:is GPVI the central receptor[J] ? Blood, 2003, 102(2):449-461.
[8] Wagner CL, MascelliMA, Neblock DS, et al.Analysisof GPIIb IIIa receptor number by quantification of 7E3 bind-ing to human platelets[J] .Blood , 1996, 88(3):907-914.
[9] Ma YQ, Qin J, Plow EF.Platelet integrin alpha(IIb) beta (3):activation mechanisms[J] .J Thromb Haemost, 2007, 5(7):1345-1352.
[10] Watanabe N, Bodin L, Pandey M, et al.Mechanisms and consequences of agonist -induced talin recruitment to plate-let integrin IIbβ3[J] .J Cell Biol, 2008, 181(7):1211-1222.
[11] Cerecedo D, Stock R, Gonzalez S, et al.Modification of actin, myosin and tubulin distribution during cytoplasmic granule movements associated with platelet adhesion[J] . Haematologica, 2002, 87(11):1165-1176.
[12] Yokoyama S, Ikeda H, Haramaki N, et al.Platelet P-se-lectin plays an important role in arterial thrombogenesis by forming large stable platelet -leukocyte aggregates[J] .J Am Coll Cardiol, 2005, 45(8):1280-1286.
[13] Zhu J, Carman CV, Kim M, et al.Requirement of alpha and beta subunit transmembrane helix separation for inte-grin outside-in signaling[J] .Blood, 2007, 110(7):2475-2483.
[14] Varughese GI, Patel JV, Tomson J, et al.Effects of blood pressure on the prothrombotic risk in 1235 patients with non-valvular atrial fibrillation[J] .Heart, 2007, 93(4): 495-499.
[15] LindenMD, Furman MI, Frelinger AL III, et al.Indices of platelet activation and the stability of coronary artery disease[J] .J Thromb Haemost, 2007, 5(4):761-765.