摘要
目的 研究异丙肌苷抗多房棘球蚴的效果和可能的机制。方法和结果 研究分三个部份，实验一异丙肌苷的药效学实验。多房棘球蚴接种小鼠30天后，随机分为六组分别口服异丙肌苷0.75，1.5，3g／kg，异丙肌苷1.5g／kg+阿苯达唑40mg／kg，阿苯达唑40mg／kg和空白对照。用药45天后取血、剖检，用药组多房棘球蚴在体积和重量上均较对照组减少，囊重抑制率分别是14.2％，36.8％，39.9％，64.8％，58.9％。对各组囊重进行统计学分析，1.5，3g／kg，异丙肌苷组和联合用药组与对照组有显著差异，观察虫体组织结构，发现有损伤和破坏，原头蚴消失。实验二异丙肌苷抗多房棘球蚴的体外实验。将多房棘球蚴组织块和原头蚴放在异丙肌苷和肌苷浓度分别为1mM，2mM，4mM的RPMI1640培养基中培养9天，发现原头蚴死亡率与阴性对照没有差异。虫体组织亦未发现有明显损伤。实验三用ELISA法检测了实验一各组小鼠血清中的IL-2，IL-4和IFN-γ，发现异丙肌苷各组小鼠的IL-2，IFN-γ，与对照组比较均有升高的趋势，IL-4有降低的趋势，但未能测出统计学差异。结论 1、异丙肌苷有一定的抗多房棘球蚴的作用，与阿苯达唑合用可取得较好的效果。2、异丙肌苷在体外没有迅速直接的杀虫作用。3、异丙肌苷在多房棘球蚴感染中有一定的免疫调控作用，但这种作用较弱。
Objective To investigate the effects and mechanisms of isoprinosine on echinococcus multilocularis metacestodes.Methods and Results The experiments were divided into three parts. In the first study, mice infected for 30 days with echinococcus multilocularis metacestodes were divided into six groups received a daily oral dose of isoprinosine 0.75,1.5,3g/kg , isoprinosine 1.5g/kg +albendazole 40mg/kg,
albendazole40mg/kg of body weight, and control. After 45day treatment, a reduction in the size and number of cysts with macroscopic modifications was observed. The inhibitory rates in different groups were 14.2%, 36.8%, 39.9%, 64.8% and 58.9% respectively. Comparing weight of cysts of groups, isoprinosine 1.5,3g/kg, isoprinosine 1.5g/kg +albendazole 40mg/kg, were significantly lower than control. The structural alterations included damage or destruction of the protoscoleces and partial destruction of the cyst wall. In the second study, protoscoleces/vesicles cultivation in vitro was used to evaluate the effect of isoprinosine and its main component inosine against protoscoleces/vesicles of Echinococcus. Isoprinosine and inosine were dissolved in RPMI1640. Concentration is ImM, 2mM, 4mM individually. The cultivation was maintained 9 days. As a result, we didn't find that isoprinosine and inosine had efficacy on protoscoleces/vesicles in vitro. In the third study we detected the titer of IL-2, IL-4 and IFN- γ
in serum of mice used in first study by ELISA. We found IL-2, IFN-γ in the groups of isoprinosine higher than that of control, IL-4 lower than control. But in statistic we didn't find differences. Conclusion Isoprinosine has
moderate efficiency of anti-Alveolar echinococcosis, especially compound with albendazole. Isoprinosine hasn't quick action to against Echinococcus Multilocularis metacestodes in vitro. Isoprinosine has moderate immunomodulation function in infection of echinococcus multilocularis metacestodes.
引文
1. Wilson. J.F and Raush. R. l Alveolar hydatid dicseases. A review of clinical features of 33 cases[J].AM J Trop Med Hyg, 1980,29:1340.
2. Ammann. R.w, Hirsbrunner. R, Cotting. J, et. al. Recurrence rate after discontinuation of long-term mebendazole therapy in Echinococcosis multilocularis[J].Am J Trop Med Hyg, 1990, 43:506.
3. Jiang Cipeng. Liver alveolar echinococcosis in the northwest report of 15 patients and collective analysis of 90 cases[J]. Chin Med J, 1981,94:771.
4. Morris DL, Dykes PW, Dickson B, et al. Albendazole in hydatid disease[J]. Br Med J (Clin Res Ed), 1983, Jan 8,286(6359): 103-104.
5. Saimot AG, Meulemans A, Cremieux AC, et al. Albendazole as apotential treatment for human hydatidosis[J]. Lancet, 1983, Sep 17,2(8351):652-656.
6. Vanparijs O. chemothrapy of experimental Echinococcus multiocularis in jirds[J]. Parasitology Res 1990, 76: 238-240.
7. WHO informal working group on echinococcosis. Guidelines for treatment of cystic and alceolar echinococcosis in humans[J]. Bulletin of the World Health Organization, 1996, 74(3): 231-242.
8. Taylor DH, Morris DI, Richard KS, et. al. Echinococcus multilocularis in vivo results of therapy with albendazole and praziquantel[J]. Trans R soc Trop Med Hyg 1988, 82, 611-615.
9. Schantz PM, Brandt FH, Dickinson CM, et. al Effects of albendazole on Echinococcus multilocularis infection in the
Mongolian jirds[J]. J infectious Dis 1990,162:1403-1407.
10. Liu YH, Wang XG, Chen YT. Computerized tomography of liver in alveolar echinococcosis treated with albendazole[J]. Zhonghua Nei Ke Za Zhi, 1993, Nov, 32(11):733-735.
11．王述恒，安国顺．免疫载药脂质体治疗小鼠泡球蚴[J]．北京医科大学学报 1997，29 (2)：109-111．
12．邱加闽，Schanitzpeter，王谦，等．奥芬达唑(Oxfendazole)治疗泡球蚴病效果初步观察[J]．实用寄生虫杂志，1999，7(3)：116-119．
13. Sarciron ME, Walchshofer N, Walbaum S, et al. Increases in the effects of albendazole on Echinococcus multilocularis metacestodes by the dipeptide methyl ester (Phe-Phe-OMe) [J].Am J Trop Med Hyg, 1997, Feb;56(2):226-230.
14. M.E Sarciron, I. Delabre, S. Walbaum, et, al Effects of multiple doses of Isoprinosine on Echinococcus multilocularis metacestodes[J]. Antimicrobial Agents and chemothepapy, jan 1992,36:191-194.
15. M.E Sarciron, S. Walbaum, et, al. Echinococcus Granulosus: isoprinosine Treatment of the metacestode stage[J] Am J Trop Med Hyg 1993,49:658-665.
16．唐维高，邓元伟，彭崇莹．抗病毒-免疫调节剂异丙肌苷的合成[J]．华西药学 1987，2：95-96．
17．曹可芬．免疫调节剂的药效学研究[J]．国外医学合成药生化药制剂分册 1988，3：136-138．
18．黄少慧．新型免疫调节剂-异丙肌苷[J]．新药与临床 1989，2：70-73．
19. Singh. H, Herndon. D.N, Effect of isoprinosine on lymphocyte proliferation and natural killer cell activity following thermal injury[J].Immunopharmacol-Immunotoxicol. 1989; 11(4): 631-644
20. Ghram A, Reddy PG, Blecha F, Minocha H C. Effects of bovine respiratory disease viruses and isoprinosine on bovine leukocyte function in vitro[J]. Vet-Microbiol. 1989 Aug; 20(4): 307-314.
21. Pedersen B K, Tvede. N, Diamant. M et. al. Effects of isoprinosine treatment of HIV-positive patients on blood mononuclear cell subsets, NK- and T-cell function, tumour necrosis factor, and interleukins 1, 2, and 6[J]. Scand-J-Immunol. 1990 Dec; 32(6): 641-649.
22. Tekgul H, Tutuncuoglu S, Kutukculer N, aizdarer G, et.al. Lymphocyte subsets and inflammatory mediators in patients with subacute sclerosing panencephalitis[J]. J-Child-Neurol. 1999 Jul; 14(7): 418-421.
23．史大中．包虫感染的免疫学新进展[J]．中国寄生虫病防治杂志1999，3：231-232．
24．史大中．包虫病治疗的新进展[J]国外医学寄生虫病分册 2001，5：231-232．
25. Gao. jianning, Zhai. zili. Advances in experimental studies on albendazole and other drugs against hydatid disease[J].中国寄生虫学与寄生虫病杂志 1991, 3:178-183.
26. Delabre I. The susceptibility of the Mongolian gerbil and the OF1 mouse strain to Echinocaccus multilocularisultrastructrual aspects of the cysts [J]. Int J Parasitology, 1987,17(3):773.
27．王静珍，孙厚英．泡球蚴病动物模型的研究[J]．地方病通报 1988，3(1)：37．
28．李维新，裴明，张国才．多房棘球蚴在不同中间宿主体内引起的病理变化[J]．地方病通报 1987，17(4)：36-39．
29．刘瑾厚，李鸿江，徐秦，等．小白鼠泡状棘球蚴病化疗效果形态学指标的探索[J]．新疆医学院学报 1995，18(4)：1-4．
30．刘章锁，杨文光，温浩，等．阿苯达唑脂质体对泡状棘球蚴作用的病理形态学观察[J]．新疆医学院学报 2000，23(4)：291-293．
31．盛迎伦，郑庆璇，郭风英，等．阿苯达唑和吡喹酮治疗沙鼠泡球蚴病的病理学观察[J]．中国寄生虫学与寄生虫病杂志 1994，12(3)178-181．
32．高劲松，刘约翰，王小根，等．阿苯达唑治疗沙鼠泡球蚴感染的实验观察[J]．地方病通报 1998，13(1)：11—13．
33. Casaso N F Rodrigez C. Ultrastructural study of in vitro larvaldeveiopnent of Echinococcus granulosus protoscoleces[J]. Int Journal for parasitology. 1989, 19(1) 21-28.
34．肖树华，尤纪青，柴君杰，等．抗包虫药物的实验研究—吡喹酮体外抗细粒棘球蚴原头节的作用[J]．地方病通报1987，2(2)43-49．
35. Lawton P, Walchshofer N, Sarciron M E. In vitro effects of isoprinosine and a dipeptide methyl ester on Echinococcus multilocularis protoscoleces[J]. Journal of Helminthology, 2001 75(3):251-257.
36．张京元，刘波，李燕宾，等．泡球蚴病和棘球蚴病患者血清IgG、IgA和IgM水平的比较[J]．中国寄生虫学与寄生虫病杂志1990，8(1)：38-40．
37．李富荣，曹和洵，蒋次鹏，等．循环抗体和循环抗原检测对泡球蚴病的疗效评价[J]．中国寄生虫病防治杂志1994，7(3)：194—196．
38. Wen H, Bresson-Hadni S, Vuitton DA, et al. Analysis of IgG subclass in the serum antibody responses of alveolar echinococcosis patients after surgical treatment and chemotherapy as an aid to assess E.g. outcome[J]. Trans R Soc Trop Med Hyg, 1995,89:692-697.
39. Rigano R, Profumo E, Teggi A, et al. Production of IL-5 and IL-6 by PBMC from patients with E, granulosus[J]. Clin Exp Immunol, 1996,105:456-459.
40. Rigano R Profumo E, Felice G, et al. invitro production of cytokines by PBMC from hydatid patients[J]. Clin Exp Immunol, 1995,99:433-439.
41. Emery L, Liance M, Deriaud E, et al. Characterization of T-cell immune responses of E. multilocularis-infected C_(57)B/6J mice[J], parasit IMMunol, 1996,18:463-472.
42. Sturm D, Menzel J, Gosttein B, et al. IL-5 is the predominant cytonine produced by PBMC in alveolar Echinococcosis[J]. Infect Immund, 1995,63:1688-1679.
43. Vuitton DA, Godot V, HarragaS, et al. Role of IL-10 in human alveolar echinococcosis[J]. J Allergy Clin Immunol, 1997, 9:S283.
44. Hadden JW.T-cell adjuvants[J].Int J Immunopharmacol 1994 Sep;16(9):703-710.
45. Cillari E, Dieli M, Lo Campo P, Sireci G, et al. Protective effect of isoprinosine in genetically susceptible BALB/c mice infected with Leishmania major[J].Immunology 1991 Sep;74(1):25-30.
46. Milano S, Dieli M, Millott S, et al. Effect of isoprinosine on IL-2, IFN-gamma and IL-4 production in vivo and in vitro[J].Int J Immunopharmacol 1991;13(7):1013-1018.
47. Pedersen BK, Tvede N, Diamant M, et al. Effects of isoprinosine treatment of HIV-positive patients on blood mononuclear cell subsets, NK- and T-cell function, tumour necrosis factor, and interleukins 1, 2, and 6[J].Scand J Immunol 1990 Dec; 32 (6): 641-649.
48. Barasoain I, Rejas MT, Ojeda G, et al. In vivo effect of isoprinosine on interleukin-2 production, lymphocyte mitogenesis and NK activity in normal and cyclophosphamide immunosuppressed mice[J], Int J Immunopharmacol 1986; 8(5):
509-515.
49. Binderup L. Effects of isoprinosine in animal models of depressed T cell function[J]. Int. J. Immunnol. 1985,137.
50．王松，许晏，朱明，等．继发性细粒棘球蚴病小鼠的细胞因子动态研究[J]．地方病通报2002，17(4)：8-11．
51. Pfister T, Schhad V, Frank W. IFN-γ in experiment echinococcosis [J].J interferon Res, 1989, 9(3) 136-138.
52. Schimid M, Samonigg R, Stoger H, et al. Use of IFN-γand mebendazole to stop the progression of alveolar hydatid disease[J].Ciln infect DiS, 1995, 20(6):1543-1546.
53．王大坤，细胞因子在寄生虫病中的治疗应用[J]．国外医学寄生虫分册，1992，19(6)：243-246．