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Praziquantel에 의한 간흡충(Clonorchis sinensis) 미세구조의 변화에 관한 연구

Other Titles
 Studies on the ultrastructural changes of clonorchis sinensis treated by praziquantel 
Authors
 차인환 
Issue Date
1982
Description
의학과/석사
Abstract
[한글] 우리나라 주요 하천을 중심으로 전국적으로 분포되어 있는 간흡충(Clonorchis sinensis)의 구충에 많은 약제가 연구되어 오고 있으나 구충효과와 그 부작용에 많은 문제점들이 제기되고 있다. 최근에 Borgers등(1975 a,b)은 benzimidazole 유도체인 mebendazole을 조충유충에, Soh 및 Min(1977), Soh등(1977)은 flubendazole을, Soh등(1979)은 praziquantel을 간흡충 감 염동물과 인체 간흡충증에 투여하여 구충효과가 있는 것으로 보고하고 있다. 그러나 아직까지 그 구충기전에 관하여 확실히 알려진 바가 없다. Jeong(1968), Kim(1968), Inatomi(1968), Kang(1968), Soh및 Min(1977), Jeong(1979)과 Fujino 및 Ishii(1979)는 간흡충의 미세구조에 대한 연구관찰을 보고한 바 있고, Moon등(1980)과 Kim등(1982)은 praziquantel을 간홉충에 감염된 실험동물에 투여시 간흡충의 미세구조의 변화를 관찰한 바 있으나 인체감염 간흡충의 미세구조의 관찰과 praziquantel의 구충기전에 관한 연구는 아직까지 보고된 바 없다. 본 실험연구는 간흡충증 환자에 있어서 praziquantel을 투여하여 시기별로 얻은 충체를 전자현미경을 이용하여 본 약제에 의한 인체기생 간흡충의 미세구조의 단계적 변화를 규명하고저 시행하였다. 본 실험에서는 간흡충 성충을 얻기 위하여 수술처치를 요하는 간흡충 감염환자에 개복술을 시행하여 환자의 총수담관에 T-tube를 삽관한 뒤 Praziquantel 50㎎/㎏을 1일 2회 분복, 2일간 투여하고 투약 후 24시간, 48시간, 72시간 후에 T-tube를 통해 배출된 간흡 충 성충을 수집하였다. 수집된 성충의 일부는 formalin-alcohol-acetic acid-glycerin(FAAG)용액에 고정하고 acetocarmine염색을 하여 광학현미경으로 관찰하였다. 일부는 3% glutaraldehyde용액으로 고정한 후 간흡충의 구흡반부, 정소부위를 분리하여 1%e osmium tet roxide로 재고정하고 통상적인 방법에 따라 uranyl acetate와 lead citrate로 염색하여 투과전자현미경으로 관찰하였다. 전자현미경적 관찰상 정상 간흡충의 외피층은 핵이 없는 합포체(syncytium)로 되어 있으며, 그 내부에 mitochondria와 2종의 분비과립이 산재하여 있으며, 외피층은 상피세포의 연장인 protoplasmic microtubule로 연결되어 있었다. 상피세포내에는 잘 발달된 RER, Golgi체, mitochondria 및 분비과립들이 관찰되었다. 정상 정소는 그 내강에 정조세포, 정모세포, 정세포 및 정자들이 조밀하게 들어 있었으며, 분화과정중의 생식세포들도 관 찰되었다. 정자의 꼬리부분에는 2개의 축사(axofilament)와 한쌍의 microtubularset가 있음이 관찰되었으며, 두부에서는 acrosome을 관찰할 수 없었다. 이러한 소견들은 실험적으로 동물에 감염시킨 간흡충의 미세구조와 동일하였다. Praziquantel 투여군에서 약물에 의한 조직의 퇴행성변화는 약물투여 24시간 후부터 나타나기 시작하여 그 정도가 시간이 지남에 따라 심화되었음을 관찰할 수 있었다. 즉, 외피층의 mitochondria의 변성, 분비과립의 공포화, 기저막의 융합 및 외피층 자체의 공포 화가 관찰되었고 상피세포내 mitochondria의 cristae의 소실, 분비과립의 변성 및 RER의 확장을 관찰할 수 있었다. 정소에서는 외피층의 퇴행성변화보다 현저하고 민감한 퇴행성 변화를 보였으나, 이미 생성된 정자는 특별한 형태학적 변화를 관찰할 수 없었다. 이상의 결과로 미루어 praziquantel은 간흡충의 단백질대사 및 합성의 장애를 초래하여 간흡충을 사멸케 하는 것으로 생각된다. Studies on the Ultrastructural Changes of Clonorchis sinensis Treated by Praziquantel. In Hwan Cha, M.D. Department of Medical Science The Graduate School, Yonsei University (Directed by Professor Keun-Tae Lee, M.D.) There are many problems in the efficacy and side effects of many drugs against Clonorshis sinensis, which is widely distributed mainly along the big rivers in Korea. Recently praziquantel(Soh et al, 1979) as a derivative of isochinoline-pyrazine was reported as a promising drug in the eradication of Clonorshis sinensis experimentally and clinically However its mechanism of action on the parasite has not been clearly demonstrated in the view of pharmacodynamics & ultrastru-ctural changes and that other studies have been carried out on the experimental animals. Therefore this study was undertaken to observe ultrastructural changes of Clonorshis sinensis infected in the human. Clonorchiasis patient who needed operative treatment was selected and he was undergone exploratory laparotomy with T-tube insertion into the common bile duct. Divided dose of 50㎎/㎏/day of praziquantel was administered for 2 days. Adult worms were collected from the bile juice which was drained from the T-tube on the 1st, 2nd and 3rd day after praziquantel administration. Under the electron microscopic observation the liver fluke in the untreated man showed normal integument which was composed of anucleated syncytium where mitochondria, round and rod-shpaed secretory granules were scattered and matrix was connected, through the protoplasmic microtubules, with epidermal cells. Well-developed rough endoplasmic reticulums, Golgi-complexes, mitochondria and numerous secretory granules in the epidermal cells were observed. In the testis which was filled with many sperms and cells of various stages in spermatogenesis, the sperm had one pair of microtubular set in the part of tail, but the acrosome was not found in the part of head. In the praziquantel treated group, the drug acted generally on the all organs, and induced remarkable degenerative changes. In the cuticle, there happened flattening of protoplasmic membrane, degenerative changes of mitochondria, vacuolization of secretory granules, fusion of the basement membrane, and appearance of homogenous low electron-density area in the nucleus and cytoplasm of the epidermal cells. In the testis more marked and more sensitive degenerative changes were noted than in the cuticle, but previously-formed sperms did not show any specific remarkable morphologic changes. Conclusively the results suggest that praziquantel interferes with normal metabolism and synthesis of protein to give rise to degerative changes in all organs of the worm.
[영문] There are many problems in the efficacy and side effects of many drugs against Clonorshis sinensis, which is widely distributed mainly along the big rivers in Korea. Recently praziquantel(Soh et al, 1979) as a derivative of isochinoline-pyrazine was reported as a promising drug in the eradication of Clonorshis sinensis experimentally and clinically However its mechanism of action on the parasite has not been clearly demonstrated in the view of pharmacodynamics & ultrastru-ctural changes and that other studies have been carried out on the experimental animals. Therefore this study was undertaken to observe ultrastructural changes of Clonorshis sinensis infected in the human. Clonorchiasis patient who needed operative treatment was selected and he was undergone exploratory laparotomy with T-tube insertion into the common bile duct. Divided dose of 50㎎/㎏/day of praziquantel was administered for 2 days. Adult worms were collected from the bile juice which was drained from the T-tube on the 1st, 2nd and 3rd day after praziquantel administration. Under the electron microscopic observation the liver fluke in the untreated man showed normal integument which was composed of anucleated syncytium where mitochondria, round and rod-shpaed secretory granules were scattered and matrix was connected, through the protoplasmic microtubules, with epidermal cells. Well-developed rough endoplasmic reticulums, Golgi-complexes, mitochondria and numerous secretory granules in the epidermal cells were observed. In the testis which was filled with many sperms and cells of various stages in spermatogenesis, the sperm had one pair of microtubular set in the part of tail, but the acrosome was not found in the part of head. In the praziquantel treated group, the drug acted generally on the all organs, and induced remarkable degenerative changes. In the cuticle, there happened flattening of protoplasmic membrane, degenerative changes of mitochondria, vacuolization of secretory granules, fusion of the basement membrane, and appearance of homogenous low electron-density area in the nucleus and cytoplasm of the epidermal cells. In the testis more marked and more sensitive degenerative changes were noted than in the cuticle, but previously-formed sperms did not show any specific remarkable morphologic changes. Conclusively the results suggest that praziquantel interferes with normal metabolism and synthesis of protein to give rise to degerative changes in all organs of the worm.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/117124
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2. 학위논문 > 1. College of Medicine (의과대학) > 석사
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