臺灣博碩士論文加值系統

Ivermectin已被廣泛應用於哺乳動物的體內外寄生蟲之防治，亦有研究指出ivermectin具有抑制腫瘤細胞生長而導致其細胞凋亡 (apoptosis) 或是死亡之特性，而其調控機制可能是藉由抑制目標細胞細胞膜上的P-glycoprotein或是γ-aminobutyric acid (GABA) receptor/chloride channel。動物在性成熟之後，睪丸組織即進行週期性的細胞分裂與分化；而在雌性動物的卵巢組織中，卵子受到雌性激素影響進而發育和排卵。因而探討ivermectin藥物是否會對睪丸組織的造精作用或是卵巢功能造成影響，在繁殖學是十分重要的課題。本研究以小鼠 (Mice) 為動物模式來探討ivermectin對哺乳動物生殖系統之影響，對雄鼠以直接睪丸內注射ivermectin 13 mg/kg，每週1次且分別投予1至6週，每組4隻；另外亦利用不同劑量 (0, 0.33, 3.3及33 mg/kg/week) 連續6週皮下注射，每組6隻，以觀察是否對其造精功能造成損害。另外直接睪丸注射投予ivermectin 13 mg/kg，每週一次且2週、並等待9週觀察其生殖系統損害是否具有回復性。結果顯示直接睪丸內注射ivermectin與高劑量皮下給予均會造成生精小管結構異常 (可見細胞壞死、變性或是細胞排列鬆散)，而計算其造精指數發現其造精指數均低於3.5 (低於3.5即為不孕)，顯示其會對造精作用造成顯著的影響。而回復性試驗方面，可發現生精小管結構異常部份 (生精小管凝固性壞死，生精小管內的spermatogonia及spermatocyte細胞變性、凋亡及壞死等) 皆可見有回復之情形。綜合上述之結果，當在小鼠以直接睪丸內注射或是高劑量 (3.3及33 mg/kg/s.c./week) 皮下給予ivermectin時，可以造成雄鼠造精功能的抑制，且其抑制作用是可回復的。

Ivermectin has been widely used on parasites control in mammals. Interestingly, some studies suggest that the ivermectin could inhibit tumor cell growth and further lead to apoptosis or death, through inhibition the P-glycoprotein or γ-aminobutyric acid (GABA) receptor / chloride channel of target cell membrane. In sexually mature animals, the spermatogenesis in testis and oocyte development in ovary are periodic processes. Hence, the effect of ivermectin in mammal reproduction functions is needed to be elucidated. The aim of this study is to evaluate the effect of ivermectin on reproductive system in mice. In male mice, experiment 1 was conducted a consecutive by direct intratesticular injection ivermectin 13 mg/kg/week for 1 - 6 weeks in different groups. Experiment 2 was conducted by subcutaneous injection with different doses (0, 0.33, 3.3 and 33 mg/kg/week) for 6 weeks in different groups. Experiment 3 was conducted by direct testicular injection of ivermectin 13 mg/kg/week for 1 week and 2 weeks, and further to evaluate the recovery process of spermatogenesis at 9th week after injection. The result of this study, ivermectin caused spermatogenetic tubule structural abnormalities (necrosis or degeneration), and significantly decreased the spermatogenesis index and the index value was lower than 3.5 (< 3.5 indicates infertility) when direct intratesticular injection or high doses of subcutaneous injection. In the recovery experiment in the male mice, the results showed that abnormal structural in spermatogenetic tubule could partly recover. According to the above results, injection of ivermectin could damage the male reproductive system function (coagulation necrosis within the seminiferous tubule, degeneration, apoptosis and necrosis of spermatogonia and spermatocyte), but the inhibitory effect could be recovered.

目錄……………………………………………………………………Ⅰ
表次……………………………………………………………………Ⅲ
圖次……………………………………………………………………Ⅴ
中文摘要………………………………………………………………Ⅷ
英文摘要………………………………………………………………Ⅹ
前言……………………………………………………………………XII

第一章　文獻探討………………………………………………………1
第一節 　Ivermectin之藥理特性…………………………………1
第二節 　Ivermectin對寄生蟲之防治……………………………2
第三節 　Ivermectin在小鼠體內分佈之情況……………………4
第四節 　Ivermectin對腫瘤多重抗藥性之抑制作用……………5
第五節 　Ivermectin對腫瘤的抑制作用…………………………6
第六節 　Ivermectin對生殖系統之影響…………………………7
第七節 　睪丸的造精作用…………………………………………9
第八節 　睪丸的造精指數………………………………………11
第九節 　第九節 TUNEL labeling原理………………………12
第十節 　研究動機………………………………………………13


第二章　材料與方法…………………………………………………14
第一節 　實驗藥品及實驗動物…………………………………14
第二節 　實驗方法………………………………………………15


第三章　結果…………………………………………………………25
第一節 　Ivermectin對小鼠雄性生殖系統毒性試驗……………25
第二節 　礦物油對小鼠雄性生殖系統毒性試驗………………33
第三節 　Ivermectin對小鼠雄性生殖系統毒性回復試驗………38
第四節 　Ivermectin皮下投予對小鼠雄性生殖系統之影響……44
第五節 　TUNEL labeling………………………………………52


第四章　討論…………………………………………………………57


第五章　參考文獻……………………………………………………74

參考文獻

1. 林桂賢。抗性腺釋放內泌素一型疫苗於犬隻繁殖能力之研究。國立中興大學獸醫學碩士論文。2003
2. 郭菁揚。應用indenopyridine hydrochloride在雄犬與大白鼠不可逆性抑制造精作用之研究。國立中興大學獸醫學碩士論文。2004
3. Allen DG, Dowling PM, Smith DA, Pasloske K, Woods JP. Hand book of veterinary drugs, 3th ed. Lippincott Williams & Wilkins, 2005.
4. Al-Azzam SI, Fleckenstein L, Cheng KJ, Dzimianski MT, McCall JW. Comparison of the pharmacokinetics of moxidectin and ivermectin after oral administration to beagle dogs. Biopharm Drug Dispos 28: 431-438, 2007.
5. Amann RP. Endocrine changes associated with onset of spermatogenesis in Holstein bulls. J Dairy Sci 66: 6-22, 1983.
6. Badi E, Luconi M, Bonaccorsi L, Muratori M, Forti G. Intracelluar events and signaling pathways involved in sperm acquisition of fertilizing capacity and acrosome reaction. Front Biosci 5: 110-123, 2000.
7. Burg RW, Miller BM, Baker EE, Birnbau M J, Currie SA, Hartmanr R, Kong YL, Monaghanrl L, Olson G, Putter I, Tunac JB, Wallick H, StapleyeE, Oiwa R, Omura S. Avermectins, new family of potent anthelmintic agents: producing organism and fermentation. Antimicrob Agents Chemother 15: 361-367, 1979.
8. Börkü MK, Atalay O, Kibar M, Cam Y, Atasever A. Ivermectin is an effective treatment for bovine cutaneous papillomatosis. Res Vet Sci 83: 360-363, 2007.
9. Calogero AE, Burrelo N, Ferrara E, Hall J, Fishel S, D’Agata R. γ-aminobutyric acid (GABA) A and B receptors mediated the stimulatory effects of GABA on human sperm acrosome reaction: intraction with progesterone. Fertil Steril 71: 930-936, 1999.
10. Chang CL, Fung HP, Lin YF, Kuo CY, Chien CW. Indenopyridine hydrochloride induced testicular spermatogenesis failure with high seminal alkaline phosphatase levels in male dog. Biol Pharm Bull 25: 1097-1100, 2002.
11. Chang LZ, Sun X, Shen S, Lei C, Liu H. Impurity effects on the crystallization of avermectin B1a. J Cryst Growth 291: 448-454, 2006.
12. Clermont Y. Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal. Physiol Rev 52: 198-236, 1972.
13. DrinyaevVA, Mosin VA, Kruglyak EB, Novik TS, Sterlina TS, Ermakova NV, Kublik LN, Levitman MK, Shaposhnikova VV, Korystov YN. Antitumor effect of avermectins. Eur J Pharmacol 501: 19-23, 2004.
14. El-Nahas AF, El-Ashmawy IM. Effect of ivermectin on male fertility and its interaction with P-glycoprotein inhibitor (verapamil) in rats. Environ Toxicol Pharmacol 26: 206-211, 2008.
15. Elbetieha A, Da’as SI. Assessment of antifertility activities of abamectin pesticide in male rats. Ecotoxicol Environ Saf 55: 307-313, 2003.
16. Geyer J, Gavrilova O, Petzinger E. Brain penetration of ivermectin and selamectin in mdr1a,b P-glycoprotein- and bcrp- deficient knockout mice. J Vet Pharmacol Ther 32: 87-96, 2009.
17. Gill JH, Kerr CA, Shoop WL, Lacey E. Evidence of multiple mechanisms of avermectin resistance in Haemonchus contortus-comparison of selection protocols. Int J Parasitol 28: 783-789, 1998.
18. Gokbulut C, Bilgili A, Hanedan B, Aksit D, Aksoy AM, Turgut C. Sex-related plasma disposition of ivermectin following pour-on administration in goats. Vet Parasitol 162: 342-345, 2009.
19. González CA, Sahagún PAM, José Diez Liébana M, Martínez NF, Vega MS, Vieitez JJ. The pharmacokinetics and metabolism of ivermectin in domestic animal species. Vet J 179: 25-37, 2009.
20. Hanna REB, Cromie L, Taylor SM, Couper A. The effect of a parenteral ivermectin/ closantel injection on the growth and reproductive development of early immature Fasciola hepatica in cattle. Vet Parasitol 142: 78-90, 2006.
21. Hoerauf A, Mand S, Volkmann L, Büttner M, Marfo-Debrekyei Y, Taylor M, Adjei O, Büttner DW. Doxycycline in the treatment of human onchocerciasis: kinetics of Wolbachia endobacteria reduction and of inhibition of embryogenesis in female Onchocerca worms. Microbes Infect 5: 261-273, 2003.
22. Kenny PJ, Vernau KM, Puschner B, Maggs DJ. Retinopathy associated with ivermectin toxicosis in two dogs. JAVMA 233: 279-284, 2008.
23. Knab J, Darge K, Bűttner DW. Immunohistological studies on macrophages in lymph nodes of onchocerciasis patients after treatment with ivermectin. Trop Med Int Health 2: 1156-1169, 1997.
24. Korystov YN, Ermakova NV, Kublik LN. Levitman, Shaposhnikova VV, Mosin VA, Drinyaev VA, Kruglyak EB, Novik TS, Sterlina TS. Avermectins inhibit multidrug resistance of tumor cells. Eur J Pharmacol 493: 57-64, 2004.
25. Mealey KL. Adverse drug reactions in herding-breed dogs: the role of P-glycoprotein. Compend Contin Educ Pract Vet 28: 23-33, 2006.
26. Mercier P, Branco FA, Sapper MF, White CR. Evaluation of the safety of ivermectin-praziquantel administered orally to pregnant mares. AJVR 64: 1221-1224, 2003.
27. Montasser AA, Gadelhak GG, Tariq S. Impact of ivermectin on the ultrastructure of the testis of Argas (Persicargas) persicus (Ixodoidea: Argasidae). Exp Appl Acarol 36: 119-129, 2005.
28. Parkinson GW, Hike KE. Bicycle helmet assessment during well visits reveals severe shortcomings in condition and fit. Pediatrics 112: 320-323, 2003.
29. Paul AJ, Hutchens DE, Firkins LD, Keehan CM. Effects of dermal application of 10.0% imidacloprid-0.08% ivermectin in ivermectin- sensitive Collies. AJVR 65: 277-278, 2004.
30. Paul AJ, Tranquilli WJ, Hutchens DE. Safety of moxidectin in avermectin-sensitive Collies. AJVR 61: 482-483, 2000.
31. Plumb DC. Plumb’s veterinary drug handbook, 6th edi. Blackwell Publish 763-770, 2008.
32. Rao UR, Chandrashekar R, Subrahmanyam D. Effect of ivermectin on serum dependent cellular interactions to Dipetalonema viteae microfilariae. Trop Med Parasitol 38: 123-127, 1987.
33. Sabeur K, Edwars DP, Meizel S. Human sperm plasma membrane progesterone receptor(s) and the acrosome reaction. Biol Reprod 54: 993-1001, 1996.
34. Sarasola P, Jernigan AD, Walker DK, Castledine J, Smith DG, Rowan TG. Pharmacokinetics of selamectin following intravenous, oral and topical administration in cats and dogs. J Vet Pharmacol Ther 25: 265-272, 2002.
35. Shoop WL, Morzik H, Fisher MH. Structure and activity of avermectins and milbemycins in animal health. Vet Parasitol 59: 139-156, 1995.
36. Uhlír J. Effect of ivermectin on the development of serum antibody activity in rabbits infested with Psoroptes cuniculi (Acari: Psoroptidae). Folia Parasitol 38: 79-82, 1991.
37. Van der Heyden S, Gheuens E, DeBruijn E, Van Oosterom A, Maes R. P-glycoprotein: clinical significance and methods of analysis. Crit Rev Clin Lab Sci 32: 221-264, 1995.
38. Villanueva I, Diez MJ, García JJ, Fernández MN, Sahagún AM, Sierra A, Sierra M. Effect of first-pass hepatic metabolism on the disposition of levamisole after intravenous administration in rabbits. Am J Vet Res 64: 1283-1287, 2003.
39. Whitsett JM, Noden PF, Cherry J, Lawton AD. Effect of transitional photoperiods on testicular development and puberty in male deer mice (Peromyscus maniculatus). J Reprod Fertil 72: 277-286, 1984.
40. Wistrom CA, Meizel S. Evidence suggesting involvement of a unique human sperm steroid receptor/Cl-channel complex in the progesterone-initiated acrosome reaction. Dev Biol 159: 679-690, 1993.
41. Wu JT, Chiang KC, Cheng FP. Expression of progesterone receptor(s) during capacitation and incidence of acrosome reaction induced by progesterone and zona proteins in boar spermatozoa. Anim Reprod Sci 93: 34-45, 2006.