肝吸蟲의 經口感染과 代謝産物로 感作된 햄스터의 腹腔渗出細包와 血淸의 肝吸蟲 感染에 대한 免疫應答

崔東翊,安斗洪,權五永 慶北大學校 醫科大學 1990 慶北醫大誌 Vol.31 No.1

肝吸蟲 被襄幼蟲의 經口感染과 肝吸蟲 成蟲의 代射産物로 感作한 近交系 golden 햄스터의 腹腔渗出細胞와 血淸을 함께 recipient 햄스터의 腹腔內注入하였을 때 免疫이 移入되는지 實驗하였다. Donor 햄스터는 5마리씩 2群으로 나누어 第Ⅰ群에는 肝吸蟲 被襄幼蟲 20마리씩을 經口感染시켰다. 第Ⅱ群에는 肝吸蟲 成蟲의 代射産物과 Freund's incomplete adjuvant의 同量 混合物 0.1㎖ 발바닥에 2週間隔으로 2回 皮內注射하였다. 2週後에 痲醉下에 腹腔渗出細胞 5×10 exp(5)과 血淸 0.5㎖를 採取하여 함께 recipient 햄스터의 腹腔內注入하였다. Recipient 햄스터는 5마리씩 3群으로 나누어 第Ⅰ群 햄스터에는 第Ⅰ群 donor 햄스터의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.5㎖를, 第2群 햄스터에는 第Ⅱ群 donor 햄스터의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.5㎖를, 第3群 對照햄스터에는 非感作 腹腔渗出細胞 5×10 exp(5)과 血淸 0.5㎖를 함께 전햄스터에 肝吸蟲 被襄幼蟲 20마리씩 challenge 經口感染시켜 第15日부터 EpG를 計算하였으며 challenge후 제50일에 全햄스터를 屠殺하여 對照群 햄스터의 負荷蟲體數와 脾臟當 plaque 形成細胞數를 其準으로 하여 感作群 햄스터와 有意成을 檢定하여 免疫의 移入與否를 判定하였다. Challenge 感染後 全群의 recipient 햄스터에서 第16日에 formalin-ether 集卵法으로 肝吸蟲卵이 發見되었고 第17日에 Eggs per Gram의 계측이 가능하였다. EpG는 對照群 第3群 recipient 햄스터에 比하여 感作群인 第2群 recipient햄스터에서 第7日 및 第50日에 적었으나 相互間에 有意的 差를 認定할 수 없었다. 第2群 recipient 햄스터의 負荷蟲體數는 第3群 햄스터의 그 數에 비하여 有意的으로 적었으나 第1群 햄스터와의 사이에는 有意的 差가 없었다. 脾臟에서 plaque 形成細胞는 第2群 recipient햄스터에서만 少數檢出할 수 있었다. 以上의 實驗結果 第Ⅱ群 donor 햄스터의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.5㎖을 腹腔內注入한 recipient 햄스터에 免疫이 移入됨을 알았다. In order to determine the effect of peritoneal exudate cell (PEC) and serum on the transfer of immunity against Clonorchis sinensis, the donor golden hamsters were senstitized as following two methods. The donor hamsters were divided into two groups. One group (Group Ⅰ) was infected orally with the metacercariae of C. sinensis and the other group (Group Ⅱ) was sensitized with two injections of the admixture of the metabolic products of adult C. sinensis and Freund's incomplete adjuvant into footpad at two weeks interval. Two weeks after sensitization, the PEC serum were collected from the hamsters under deep anesthesia. Recipient hamsters were divided into three groups. The hamsters of Group Ⅰ were injected intraperitoneally(IP) with 5×10 exp(5) PEC and 0.5 ㎖ of serum from Group Ⅰ donor hamsters, and those of Group 2 were injected IP with 5×10 exp(5) PEC and 0.5㎖ of serum form Group Ⅱ donor hamsters. The control hamsters of Group 3 were injected IP with 5×10 exp(5) PEC and 0.5㎖ of serum from nonsensitized controls. Seven days after primary sensitization, the recipient hamsters were challenged orally with 20 metacercariae of C. sinensis and Eggs per Gram (EpG) of fecal samples were counted from 15 to 50 days after challenge. The recipient hamsters were killed 50 days after challenge, and the transfer of immunity to the recipient hamsters was determined by significant differences in worm burdens and plaque cells per spleen between sensitized and non-sensitized groups. The eggs of C. sinensis in the all groups appeared in 16 days by the formalin-ether sedimentation and EpG appeared in 17 days by Stoll's egg counting techniques after challenge. In 47 and 50 days after challenge, the EpG in the sensitiaed group were lesser than those in nonsensitized group. However, no significant difference was observed in EpG pattern between the sensitized and non-sensitized groups. There were some plaque forming cells in the Group 2 hamsters given immune PEC and serum. In the light of these results, it is found that the donor hamsters of Group Ⅱ transfered immunity to the recipient hamsters.

肝吸蟲代謝産物과 蟲體構成物로 感作한 햄스터에서 免疫의 移入

崔東翊,朴武吉 慶北大學校 醫科大學 1987 慶北醫大誌 Vol.28 No.3

肝吸蟲의 代謝産物과 蟲體構成物로 感作한 近交系 golden 햄스터의 腹腔渗出細胞, 脾臟細胞 및 血淸이 recipient햄스터에 免疫을 移入하는지 究明하였다. Donor 햄스터는 5마리씩 2群으로 나누어 第Ⅰ群에는 0.1㎖의 代謝産物과 Freund's incomplete adjuvant의 同量 混合物을, 第Ⅱ群에는 0.1㎖의 蟲體構成物과 adjuvant의 同量 混合物을 ㄹootpad에 2回 注射한 다음 2週後에 麻醉시켜 腹腔渗出細胞, 脾臟細胞 및 血淸을 採集하여 recipient햄스터의 腹腔內에 注入하였다. Recipient햄스터는 5마리씩 6群으로 나누어 第1群에는 第Ⅰ群 donor 햄스터의 腹腔渗出細胞 5×10 exp (5) 및 血淸 1㎖를, 第2群에는 第Ⅰ群의 脾臟細胞 2×10 exp (6) 및 血淸 1㎖를, 第3群에는 第Ⅱ群 donor 햄스터의 腹腔渗出細胞 5×10 exp (5) 및 血淸 1㎖를, 第4群에는 第Ⅱ群의 脾臟細胞 2×10 exp (6) 및 血淸 1㎖를, 第5群에는 非感作對照햄스터의 腹腔渗出細胞 5×10 exp (5) 및 血淸 1㎖를, 第6群에는 對照햄스터의 脾臟細胞 2×10 exp (6) 및 血淸 1㎖를 各各 腹腔內에 注入하여 第1次 感作하였다. 第1次 感作한 7日後 全햄스터에 肝吸蟲 被襄幼蟲 30마리를 經口 challenge感染시켜 15日부터 EpG의 計算을 하였고 challenge 感染後 50日에 모두 屠殺하여 對照群의 worm burden 및 脾臟當 plaque形成細胞數를 基準으로 하여 이들 細胞와 血淸의 注入으로 recipient햄스터에 免疫이 移入되는지 그 與否를 判定하였다. Challenge 感染後 第1, 第5 및 第6群 햄스터에서는 第15日째, 第2, 第3 및 第4群 햄스터에서는 第16日째에 肝吸蟲卵이 formalin-ether 集卵法으로 나타났으며 Eggs per Gram은 Stoll氏 計算法으로 第16日 및 第17日째부터 나타났고, 단도에서 肝吸蟲의 排出로 推定되는 EpG의 急激한 增加는 感作群인 第1, 第2 및 第4群에서는 第28日째, 第3群에서는 31日째에 나타났는데 比하여 非感作對照群인 第5 및 第6群에서는 40日 또는 43日째에 나타났다. 햄스터 단도내 worm burden은 感作群 햄스터(平均 13.2∼15.0마리)는 非感作對照群 햄스터(平均 18.4∼19.0마리)에 比하여 有意的으로 적었다. 脾臟에서 plaque形成細胞는 非感作對照群에서는 檢出할 수 없었고 感作群인 第1, 第2, 第3 및 第4群 햄스터에서는 少數 檢出할 수 있었다. 이 成績으로 미루어 보아 肝吸蟲의 代謝産物과 蟲體構成物로 感作한 햄스터의 腹腔渗出細胞 및 血淸과 脾臟細胞 및 血淸의 recipient햄스터의 腹腔內注入은 免疫이 移入됨을 나타내었다. This study was undertaken to evaluate the role of peritoneal exudate cells(PEC), spleen cells (SC) and serum in the transfer of immunity against Clonorchis sinensis in the isogenic golden hamsters. The donor hamsters (DH) were divided into two groups. One group was sensitized with twice injections of the admixture of the metabolic products of C. sinensis and Freund's incomplete adjuvant into footpads, and the other group was sensitized with the admixture of somatic constituents and the adjuvant. Two weeks after sensitization, the DH were killed by deep anesthesia and the PEC, SC and serum were collected. Recipient hamsters (RH) were divided into 6 groups. The hamsters of Group Ⅰ were injected intraperitoneally (IP) with 5×10 exp (5) PEC and 1 ㎖ of serum from Group Ⅰ DH, those of Group 2 were injected IP with 2×10 exp (6) SC and 1 ㎖ of serum from Group Ⅰ DH, those of Group 3 were injected IP with 5×10 exp (5) PEC and 1㎖ of serum from Group Ⅱ DH and those of Group 4 were injected IP with 2×10 exp (6) SC and 1 ㎖ of serum from Group Ⅱ DH. The hamsters of Group 5 were injected IP with 5×10 exp (5) PEC and 1 ㎖ of serum and those of Group 6 were adminstered IP with 2×10 exp (6) SC and 1 ㎖ of serum from non-sensitized controls. Seven days after primary sensitization, RH were challenged orally with 30 metacercariae and Eggs per Gram (EpG) of fecal samples were counted from 15 to 49 days after challenge. The RH were killed 50 days after challenge, and the transfer of immunity to the RH was estimated by significances in EpG, mean worm burdens, and plaque forming cells per spleen between sensitized and non-sensitized groups. The eggs of C. sinensis appeared in the 15th day after challenge in Groups 1,5,6 and the 16th day in Groups 2,3,4, by the formalin-ether sedimentation and EpG noted in the 16th and the 17th days by Stoll's egg counting techniques. The sudden increase of EpG, which assumed to be expelled the flukes into the intestinal tract, was encountered in the 28 days in Groups 1,2,4, and the 31st day in Group 3. Whereas, it appeared in the 40th day in Group 5 and the 43rd day in Group 6. The mean numbers of sensitized groups (RH of Groups 1,2,3,4) harbored fewer flukes than the non-sensitized groups and the differences in mean worm burdens were significant by the paired t-test. The plaque forming cells were found in the all sensitized groups, but no plaque forming cell was encountered in the nonsensitized groups. It is likely that the intraperitoneal injection of peritoneal exudate cells and serum or spleen cells and serum from the donor hamsters sensitized with the metabolic products and somatic constituents of C. sinensis caused the transfer of immunity to the recipient hamsters.

Potency of Cynomorium songaricum to prevent the regressing effects of short photoperiod in male golden hamsters

Ji Young Kwon,Hyun Ah Oh,Min Ho Jeong,Donchan Choi 한국발생생물학회 2013 한국발생생물학회 학술발표대회 Vol.2013 No.8

Golden hamsters are seasonal breeders whose reproductive activities are active during summer. Their sexual function is completely suppressed in winter. In oriental society, traditional medicines have been utilized in treating various complaints. One of them is Cynomorium songaricum (CS) whose extract has been used in treating male impotence and sexual dysfunction. We investigated the effects of aqueous CS extract on the process of spermatogenesis in golden hamsters. The animals were divided into 4 groups: long photoperiod (LP) control, short photoperiod (SP) control, and SP animals treated with low or high concentrations of CS. The animals were daily intubated with low (1.25 g/kg) or high (2.50 g/kg) concentrations of the CS aqueous extracts for 8 weeks. The control animals received the vehicle. The volume of testis was measured consecutively from the same animal at 4 week intervals. As results, the LP control animals showed large testes that indicate reproductively active function, but SP control animals displayed remarkably reduced testes that reflect inactive function. The outcomes of the reproductive activity from low concentrations of CS were not conspicuous. And the integral consequences of the reproductive activity from high concentrations of CS treatments were not significant either. But in tracing the individual animals treated with high CS extract, the effects were evidently splitted into dichotomy. In one subgroup small testes were displayed as in SP control animals, but in the other subgroup large testes were demonstrated as in LP control animals, which implies the absolute prevention of regressing activity by SP. These results suggest that the CS extract has a potency to prevent the regressing effects of SP and promotes the male fertility by strengthening the spermatogenesis in the golden hamsters.

A Study for the Expression of Melatonin Receptor Gene and Reproductive Indices in Golden Hamsters Exposed to Photoperiods

Park Donchan,Park Hyungjae,Sinae Lim,Park Changeun 한국발생생물학회 2002 발생과 생식 Vol.6 No.1

골든 햄스터의 생식활동은 광주기에 의해 조절된다. 그들의 생식능력은 여름에 왕성하고 겨울에는 퇴화한다. 송과선에서 분비되는 멜라토닌은 계절적 번식동물에서 생식활동을 중재한다. 멜라토닌 수용체가 최근에 사람을 포함하는 몇몇 동물에서 확인되었지만 골든 햄스터의 생식능력과 관련하여 알려진 바가 많지 않다. 역전사 PCR 방법을 사용하여 멜라토닌 수용체의 일부 유전자를 동정하였다(309 염기). 멜라토닌 수용체의 핵산 서열과 추론된 아미노산 서열을 보고된 다른 Reproductive activity of golden hamsters(Mesocricetus auratus) is regulated by the photoperiod. They are sexually active in summer and inactive in winter. Melatonin, a pineal hormone, has been known to mediate sexual activities in seasonal breeding animals. Melatonin receptor was recently identified in several animal species including hmm. But little has been known about it in relation to the reproductive activities of golden hamsters. By using reverse transcription polymerase chain reaction(RT-PCR) methods, a portion of the melatonin receptor gene(309 nucleotides) was identified in golden hamsters. The nucleotide sequence of the melatonin receptor and the amino acid sequence deduced were compared to those reported in other animals. Melatonin receptors were obviously detected in hypothalamus, pituitary containing pars tuberalis, blood, and spleen. Although the testicular weights and the levels of reproductive hormones were dramatically affected by photoperiods, the expression of melatonin receptor was not markedly changed by them. These results suggest that the action of melatonin in regulating reproduction might be mainly due to the affinity of melatonin receptor rather than the density fi melatonin receptor.

골든 햄스터의 생식내분비계에 미치는 멜라토닌의 영향

최돈찬,우대균,임시내 한국환경생물학회 2002 환경생물 : 환경생물학회지 Vol.20 No.3

광주기(하루 중 빛의 길이)는 골든 햄스터의 생식을 조절하는 주된 요인이다. 광주기 정보는 멜라토닌을 통하여 생식 내분비계로 전달된다. 따라서 멜라토닌이 생식에 미치는 효과를 여러 광주기에 노출시킨 햄스터에서 조사하였다. 단주기(하루 중 12시간 이하의 조명)에 노출시킨 동물들과 저녁에 멜라토닌을 주사한 동물들의 정소 무게는 현저하게 줄어들었으나, 장주기 (하루 중 12.5시간 이상의 조명)에 유지된 동물과 오전에 멜라토닌을 투여한 동물들의 정소 무게는 줄어들지 않았다. 퇴화된 정소를 조직학적으로 조사한 결과, 세정관 직경이 감소되었고,세정관내 세포수가 두드러지게 줄어들었다. 또한 생식 능력이 퇴화된 동물의 혈중 여포자극호르몬과 황체호르몬의 수준도 생식 능력을 보유하고 있는 동물에 비해 뚜렷하게 감소하였다. 멜라토닌 수용체가 역전사 polymerase chain reaction으로 등정되었고 조직특이성 또한 조사하였다 동정된 멜라토닌 수용체는 309 염기였으며,시상하부와 뇌하수체를 포함하는 다양한 장기에서 발현되었다. 생식을 조절하는 핵심 물질인 gona-dotropin releasing hormone (GnRH) 유전자의 발현 또한 동정되었다. 그러나 멜라토닌 처리와 광주기 처리는 GnRH유전자 발현에 영향을 미치지 않았다. 종합하면, 광주기의 효과는 멜리토닌을 경유하여 발휘되며,멜라토닌은 GnRH유전자의 발현보다는,생성된 GnRH의 분비에 영향을 미쳐 생식내분비계에 간접적으로 작용함을 알 수 있었다. Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin are investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod (SP, ??12hours day^-1) and injected with melatonin in the evening, but not in long photoperiod(LP, ??12.5 hours day^-1) and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

Effects of Dietary Supplement Containing Melatonin on Reproductive Activity in Male Golden Hamsters

최돈찬 한국발생생물학회 2019 발생과 생식 Vol.23 No.2

Melatonin is a pineal hormone that is synthesized and released at night under the light and dark cycles of a day. Its effects on the reproductive activities have well been established by the administration through various routes in photoperiodic animals. It was also identified in plants and named phytomelatonin. The capacity of the phytomelatonin was investigated in this investigation whether it affects the reproductive function in male golden hamster. As expected, animals housed in long photoperiod (long photoperiod, LP>12.5 hours of lights in a day) had large testes and animals kept in short photoperiod (SP≦12.5 hours of lights in a day) showed remarkably reduced testes. The dietary supplement with melatonin itself induced the complete involution of testes. Pistachios that were reported to contain a large amount of melatonin demonstrated no effects at all in male golden hamsters. These results suggest that dietary supplement containing melatonin-rich foodstuff used in this investigation may not be enough to affect the reproductive endocrine system in male golden hamsters

Effects of Dietary Supplement Containing Melatonin on Reproductive Activity in Male Golden Hamsters

Choi, Donchan The Korean Society of Developmental Biology 2019 발생과 생식 Vol.23 No.2

Melatonin is a pineal hormone that is synthesized and released at night under the light and dark cycles of a day. Its effects on the reproductive activities have well been established by the administration through various routes in photoperiodic animals. It was also identified in plants and named phytomelatonin. The capacity of the phytomelatonin was investigated in this investigation whether it affects the reproductive function in male golden hamster. As expected, animals housed in long photoperiod (long photoperiod, LP>12.5 hours of lights in a day) had large testes and animals kept in short photoperiod ($$SP{\leq_-}12.5$$ hours of lights in a day) showed remarkably reduced testes. The dietary supplement with melatonin itself induced the complete involution of testes. Pistachios that were reported to contain a large amount of melatonin demonstrated no effects at all in male golden hamsters. These results suggest that dietary supplement containing melatonin-rich foodstuff used in this investigation may not be enough to affect the reproductive endocrine system in male golden hamsters.

Continuous Melatonin Attenuates the Regressing Activities of Short Photoperiod in Male Golden Hamsters

Donchan Choi 한국발생생물학회 2013 발생과 생식 Vol.17 No.2

Golden hamsters reproduce in a limited time of a year. Their sexual activities are active in summer but inactive in winter during which day length does not exceed night time and environmental conditions are severe to them. The reproductive activities are determined by the length of light in a day (photoperiod). Melatonin is synthesized and secreted only at night time from the pineal gland. Duration of elevated melatonin is longer in winter than summer, resulting in gonadal regression. The present study aimed at the influences of continuous melatonin treatments impinging on the gonadal function in male golden hamsters. Animals received empty or melatonin-filled capsules for 10 weeks. They were divided into long photoperiod (LP) and short photoperiod (SP). All the animals maintained in LP (either empty or melatonin-filled capsules) showed large testes, implying that melatonin had no effects on testicular functions. Animals housed in SP displayed completely regressed testes. But animals kept in SP and implanted with melatonin capsules exhibited blockage of full regression by SP. These results suggest that constant release of melatonin prohibits the regressing influence of SP.

Continuous Melatonin Attenuates the Regressing Activities of Short Photoperiod in Male Golden Hamsters

최돈찬 한국발생생물학회 2013 발생과 생식 Vol.17 No.2

Golden hamsters reproduce in a limited time of a year. Their sexual activities are active in summer but inactive in winter during which day length does not exceed night time and environmental conditions are severe to them. The reproductive activities are determined by the length of light in a day (photoperiod). Melatonin is synthesized and secreted only at night time from the pineal gland. Duration of elevated melatonin is longer in winter than summer, resulting in gonadal regression. The present study aimed at the influences of continuous melatonin treatments impinging on the gonadal function in male golden hamsters. Animals received empty or melatonin-filled capsules for 10 weeks. They were divided into long photoperiod (LP) and short photoperiod (SP). All the animals maintained in LP (either empty or melatonin-filled capsules) showed large testes, implying that melatonin had no effects on testicular functions. Animals housed in SP displayed completely regressed testes. But animals kept in SP and implanted with melatonin capsules exhibited blockage of full regression by SP. These results suggest that constant release of melatonin prohibits the regressing influence of SP.

Continuous Melatonin Attenuates the Regressing Activities of Short Photoperiod in Male Golden Hamsters

Choi, Donchan The Korean Society of Developmental Biology 2013 발생과 생식 Vol.17 No.2

Golden hamsters reproduce in a limited time of a year. Their sexual activities are active in summer but inactive in winter during which day length does not exceed night time and environmental conditions are severe to them. The reproductive activities are determined by the length of light in a day (photoperiod). Melatonin is synthesized and secreted only at night time from the pineal gland. Duration of elevated melatonin is longer in winter than summer, resulting in gonadal regression. The present study aimed at the influences of continuous melatonin treatments impinging on the gonadal function in male golden hamsters. Animals received empty or melatonin-filled capsules for 10 weeks. They were divided into long photoperiod (LP) and short photoperiod (SP). All the animals maintained in LP (either empty or melatonin-filled capsules) showed large testes, implying that melatonin had no effects on testicular functions. Animals housed in SP displayed completely regressed testes. But animals kept in SP and implanted with melatonin capsules exhibited blockage of full regression by SP. These results suggest that constant release of melatonin prohibits the regressing influence of SP.