음양곽(Epimedicum Koreanum nakai)은 예부터 생식기 장애치료를 위해 사용되어 왔다. 본 연구에서는 2-브로모프로판(2-bromopropane; 2-BP)으로 유발된 생식기 장애에서 음양곽 물추출물의 생식기장애 치료효과를 평가하였다. 2-BP를 체중kg당 1,355 mg씩 28일간 피하로 투여하여 생식기 장애를 유발하였으며, 음양곽 추출물은 용량별(10, 100, 500 mg/kg)로 4주간 경구투여 하였다. 그 결과 일일정자생성(daily sperm production)은 대조군에 비하여 증가하였으나 용량의존적 감소를 나타내었으며, 정세관과 부고환관에서는 조직학적 유의한 변화를 나타내었다. 이러한 결과는 음양곽물추출이 2-BP로 유발된 생식기장애의 치료에 도움이 될 것으로 생각된다.
음양곽(Epimedicum Koreanum nakai)은 예부터 생식기 장애치료를 위해 사용되어 왔다. 본 연구에서는 2-브로모프로판(2-bromopropane; 2-BP)으로 유발된 생식기 장애에서 음양곽 물추출물의 생식기장애 치료효과를 평가하였다. 2-BP를 체중kg당 1,355 mg씩 28일간 피하로 투여하여 생식기 장애를 유발하였으며, 음양곽 추출물은 용량별(10, 100, 500 mg/kg)로 4주간 경구투여 하였다. 그 결과 일일정자생성(daily sperm production)은 대조군에 비하여 증가하였으나 용량의존적 감소를 나타내었으며, 정세관과 부고환관에서는 조직학적 유의한 변화를 나타내었다. 이러한 결과는 음양곽물추출이 2-BP로 유발된 생식기장애의 치료에 도움이 될 것으로 생각된다.
Epimedium koreanum nakai (EKN) has been used for treatment of reproductive disorders. In this study, we evaluated the effects of EKN water extract on the 2-bromopropane (2-BP) induced reproductive dysfunction. The daily sperm production (DSP), sperm counts and histological changes of reproductive or...
Epimedium koreanum nakai (EKN) has been used for treatment of reproductive disorders. In this study, we evaluated the effects of EKN water extract on the 2-bromopropane (2-BP) induced reproductive dysfunction. The daily sperm production (DSP), sperm counts and histological changes of reproductive organs were investigated after 4 weeks of EKN administration in 2-BP induced reproductive damaged rats. Although the weights of epididymis and seminal vesicle were increased dose-dependently, they were lower than control group. DSP of EKN-dosing groups were increased compared to control group. In EKN-dosing groups, the number of degenerative seminiferous tubules and the number of epididymal tubules showing epithelial cell vacuolation and decreased spermatozoa in the lumen were significantly (p < 0.01) decreased compared to control group dose-dependently. Significant (P < 0.05 or P < 0.01) increases of the number of sertoli cells, spermatogonia, patchytene spermatocytes, round spermatids and elongated spermatids were observed in EKN-dosing groups compared control group. Consequently, based on the results, EKN water extract could treat 2-BP induced reproductive damages dramatically.
Epimedium koreanum nakai (EKN) has been used for treatment of reproductive disorders. In this study, we evaluated the effects of EKN water extract on the 2-bromopropane (2-BP) induced reproductive dysfunction. The daily sperm production (DSP), sperm counts and histological changes of reproductive organs were investigated after 4 weeks of EKN administration in 2-BP induced reproductive damaged rats. Although the weights of epididymis and seminal vesicle were increased dose-dependently, they were lower than control group. DSP of EKN-dosing groups were increased compared to control group. In EKN-dosing groups, the number of degenerative seminiferous tubules and the number of epididymal tubules showing epithelial cell vacuolation and decreased spermatozoa in the lumen were significantly (p < 0.01) decreased compared to control group dose-dependently. Significant (P < 0.05 or P < 0.01) increases of the number of sertoli cells, spermatogonia, patchytene spermatocytes, round spermatids and elongated spermatids were observed in EKN-dosing groups compared control group. Consequently, based on the results, EKN water extract could treat 2-BP induced reproductive damages dramatically.
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제안 방법
All rats were injected 2-BP at dose of 1,355 mg/kg subcutaneously once a day for 28 days. In the day after last injection of 2-BP, the rats were divided into 3 EKN-dosing groups by body weights: EKN-10 (10 mg/kg EKN-dosing group), EKN-100 (100 mg/kg EKN-dosing group), EKN-500 (500 mg/kg EKNdosing group) and control group in which each of 8 consisted. Then the test article was administered to the 3-EKN-dosing groups per os using a gastric gavage once a day for 28 days while control group was given with DW in the same manner instead of the test article.
The number of seminiferous tubules showing atrophic changes or containing vacuolated sertoli cells in testis, the number of epididymal tubules showing oligospermatozoa or aspermatozoa, and the number of epididymal tubules showing epithelial cell vacuolation were calculated as the number per 100 tubules, respectively. The histomorphometry was conducted using automated image analysis (DMI-300 Image Processing; DMI, Korea) under magnification 200 of microscopy (Nikon, Japan) at 5 fields (n = 5), respectively.
In the day after last injection of 2-BP, the rats were divided into 3 EKN-dosing groups by body weights: EKN-10 (10 mg/kg EKN-dosing group), EKN-100 (100 mg/kg EKN-dosing group), EKN-500 (500 mg/kg EKNdosing group) and control group in which each of 8 consisted. Then the test article was administered to the 3-EKN-dosing groups per os using a gastric gavage once a day for 28 days while control group was given with DW in the same manner instead of the test article.
Therefore, we performed this study to investigate the effects of EKN water extract on reproductive failure induced by 2-BP in Sprague-Dawley rats.
대상 데이터
Thirty-two 8 week-old male Sprague-Dawley rats (Crj/Bgi: CD(SD)IGS: 236 ± 21 g body weight) were obtained from Oriental Bio (Seoul, Korea) and used after acclimatization for 1 week.
성능/효과
After administration of EKN extract for 28 days in reproductive damaged rats induced by 2-BP, the absolute spleen weight in EKN-500 group was significantly (P < 0.05) decreased compared to control group but no significant change was observed in relative weight.
Based on the results, it is concluded that EKN water extract dramatically treats the 2-BP induced-reproductive damages from 10 mg/kg of dose levels, the lowest levels. Although 10 mg/kg of EKN extract also inhibited the decreases of spermatogenesis, the effective dosage of EKN inhibiting both reproductive damages and decreases of spermatogenesis by 2- BP was considered as 100 mg/kg in rats because significant inhibition of spermatogenesis decreases was restricted to some stages of some cell types in 10 mg/kg dosing group.
In EKN-10 and EKN-100 groups, not only the both absolute epididymis weights were significantly (P < 0.05) decreased but the both absolute and relative seminal vesicle weights were significantly (P < 0.01 or P < 0.05) decreased compared to control group.
In the previous studies (17,36), the decreases of DSP and sperm number in 2-BP treated rats resulted from apoptosis. In this study, DSPs of EKN-dosing groups were decreased with dose-dependent pattern while this result came into conflict with histomorphological results (low damage of spermatozoa). It was supposed that DSP ability had relation to sperm count, that is, plenty of sperms stimulated negative feedback as low DSP.
In this study, significant (p < 0.01 or p < 0.05) increases of the numbers of sertoli cells, spermatogonia, preleptotene spermatocytes, leptotene spermatocytes, zygote spermatocytes, pachytene spermatocytes, round and elongated spermatids were observed in EKN-100 and EKN-500 at all stages, compared to control group.
In this study, DSPs of EKN-dosing groups were decreased with dose-dependent pattern while this result came into conflict with histomorphological results (low damage of spermatozoa). It was supposed that DSP ability had relation to sperm count, that is, plenty of sperms stimulated negative feedback as low DSP. Sperm counts of EKN-dosing groups were increased dose-dependently.
Significant (p < 0.01) increases of the number of sertoli cells, spermatogonia, preleptotene spermatocytes, leptotene spermatocytes, zygote spermatocytes, pachytene spermatocytes, round and elongated spermatids were observed in EKN-100 and EKN-500 at all stages and significant (P < 0.05 or P < 0.01) increase of those were observed in EKN-10 at all stages except stage XII compared to control group (Table 4, fig 2).
The number of degenerative seminiferous tubules and the number of epididymal tubules showing epithelial cell vacuolation and decreased spermatozoa in the lumen in EKN-dosing groups were significantly (p < 0.01) decreased compared to control group dose-dependently (Table 3, fig 1).
The platelet and white blood cell (WBC) of EKN-dosing groups were significantly (P < 0.01 or P < 0.05) decreased, respectively, and mean corpuscular volume (MCV) of EKN-500 group was decreased significantly (P < 0.01) compared to control group.
후속연구
In this study, any significant change of reproductive organ was not observed in the low dose (EKN-10) group. Therefore further investigation about it might be needed. Over the 500 mg/kg of EKN extract has anti-impotent effect against 2-BP induced-genital damage by increasing the weight of reproductive organ.
참고문헌 (40)
Blazak WF, Treinen KA, Junievicz PE. Application of testicular sperm head counts in the assessment of male reproductive toxicity. In: chapin RE, Heindel JJ, editors. Methods in toxicology: male reproductive toxicology. San Diego: Academic press. 1993: 86-94.
Chen KM, Ge BF, Ma HP, Liu XY, Bai MH, Wang Y. Icariin, a flavonoid from the herb Eimedium enhances the osteogenic differentiation of rat primary bone marrow stromal cells. Pharmazie 2005; 60: 939-942.
Chen KM, Ge BF, Liu XY, Ma PH, Lu MB, Bai MH, Wang Y. Icariin inhibits the osteoclast formation induced by RANKL and macrophage-colony stimulating factor in mouse bone marrow culture. Pharmazie 2007; 62: 388-391.
Creasy DM. Evaluation of testicular toxicity in safety evaluation studies: the appropriate use of spermatogenic staging. Toxicol Pathol 1997; 25: 119-131.
Huang J, Yuan L, Wang X, Zhang TL, Wang K. Icaritin and its glycosides enhance osteoblastic, but suppress osteoclastic, differentiation and activity in vitro. Life Sci 2007; 81: 832-840.
Hou M, Chrysis D, Nurmio M, Parvinen M, Eksborg S, Soder O, Jahnukainen K. Doxorubicin induces apoptosis in germ line stem cells in the immature rat testis and amifostine cannot protect against this cytotoxicity. Cancer Res 2005; 65: 9999-10005.
Ichihara G, Asaeda N, Kumazawa T, Tagawa Y, Kamijima M, Yu X, Konda H, Nakajima T, Kitoh J, Yu IJ, Moon YH, Hisanaga N, Takeuchi Y. Testicular and hematopoietic toxicity of 2-bromopropane, a substitute for ozone layer-depleting chlorofluorocarbons. J Occup Health 1997; 39: 57-63.
Jahnukainen K, Chrysis D, Hou M, Parvinen M, Eksborg S, Soder O. Increased apoptosis occurring during the first wave of spermatogenesis is stage-specific and primarily affects midpachytene spermatocytes in the rat testis. Biol Reprod 2004; 70: 290-296.
Jeong TC, Lee ES, Chae W, Koh WS, Kang BH, Han SS. Immunotoxic effects of 2-bromopropane in male Spraguedawley rats: a 28-day exposure study. J Toxicol Environl Health A 2002; 65: 383-394.
Kang KS, Li GX, Che JH, Lee YS. Impairment of male rat reproductive function in F1 offspring from dams exposed to 2- bromopropane during gestation and lactation. Reprod Toxicol 2002; 16: 151-159.
Kim HJ, Kang BN, Cho SW, Son HY, Jeong NS, Park SJ, Kim SH, Kim SR, Kim TH, An MY, Ryu SY. Effect of Benzo[a]pyrene, 2-bromopropane, phenol and 2,3,7,8-tetrachlorodibenzo- p-dioxin on proinflammatory cytokines gene expression by mice spleen cells. J Vet Sci 2002; 3: 247-254.
Kim HT, Kim JW, Lim MK, Jin TW, Yeo SG, Jang KH, Oh TH, Lee KW. Cytotoxic effect of Artemisia capillaris extracts on the cancer cells on in vitro. J Vet Clin 2007; 24: 367-371.
Kim Y, Jung K, Hwang T, Jung G, Kim H, Park J, Kim J, Park J, Park D, Park S, Choi K, Moon Y. Hematopoietic and reproductive hazards of Korean electronic workers exposed to solvents containing 2-bromopropane. Scand J Work Environ Health 1996; 22: 387-391.
Li GX, Kang KS, Lee YS. 2-bromopropane induced germ cell apoptosis during spermatogenesis in male rat. J Vet Med Sci 2000; 63: 373-382.
Liu WJ, Xin ZC, Xin H, Yuan YM, Tian L, Guo YL. Effects of icariin on erectile function and expression of nitric oxide synthase isoforms in castrated rats. Asian J Androl 2005; 7: 381-388.
Liu ZQ. Icariin: A special antioxidant to protect linoleic acid against free-radical-induced peroxidation in micelles. J Phys Chem A 2006; 110: 6372-6378.
Makarova MN, Pozharitskaya ON, Shikov AN, Tesakova SV, Makarov VG, Tikhonov VP. Effect of lipid-based suspension of Epimedium koreanum Nakai extract on sexual behavior in rats. J Ethnopharmacol 2007; 114: 412-416.
Matsui H, Toyoda K, Kawanashi T. Advantages of simplified quantitative morphometry using stage grouping analysis of spermatogenic cycle for evaluation of the testicular toxicity of ethylene-1, 2-dimethanesulfonate in rats. J Toxicol Pathol 1996; 9: 285-292.
Omura M, Romero Y, Zhao M, Inoue N. Histopathological evidence that spermatogonia are the target cells of 2- Bromopropane. Toxicology Letters 1999; 104: 19-26.
Pang Y, Zhang WY, Xia X, Kong LD. Effects of icariin on hypothalamic-pituitary-adrenal axis action and cytokine levels in stressed Sprague-Dawley rats. Biol Pharm Bull 2006; 29: 2399-2403.
Sha M, Cao A, Yang S, Xue Y. Determination of icariin in epimedium koreaum nakai by high performance liquid chromatography. Se Pu 1997; 15: 166-167.
Shin KH, Kang SS, Chung SG, Cho EH. Determination of icariin in Epimedii herba by high performance liquid chromatography. Kor J Pharmacogn 1989; 20: 21-24.
Son HY, Kim YB, Kang BH, Gho SW, Ha CS, Roh JK. Effects of 2-bromopropane on spermatogenesis in the Sprague- Dawley rats. Reprod Toxicol 1999; 13: 179-187.
Sun PY, Chen YJ, Wen Y, Pei YP, Liu ZH, Yao XS, Takeda T, Ogihara Y. Structure determination of korepimedoside A and korepimedoside B from Epimedium koreanum nakai. Yao Xue Xue Bao 1996; 31: 602-606.
Tian L, Xin ZC, Yuan YM, Fu J, Liu WJ, Wang LL. Effects of icariin on intracavernosal pressure and systematic arterial blood pressure of rat. Zhonghua Yi Xue Za Zhi 2004; 84: 142-145.
Wu X, Faqi AS, Yang J, Ding X, Jiang X, Chahoud I. Male reproductive toxicity and beta-luteinizing hormone gene expression in sexually mature and immature rats exposed to 2-bromopropane. Hum Exp Toxicol 1999; 18: 683-690.
Yamada T, Inoue T, Sato A, Yamagishi K, Sato M. Effects of short-term administration of alph-chlorohydrin on reproductive toxicity parameters in male Sprague-Dawley rats. J Toxicol Sci 1995; 20: 195-205.
Yu IJ, Chung YH, Lim CH, Maeng SH, Lee JY, Kim HY, Lee SJ, Kim CH, Kim TG, Lim CH, Park JS, Moon YH. Reproductive toxicity of 2-bromopropane in Sprague Dawley rats. Scand J Work Environ Health 1997; 23: 281-288.
Yu IJ, Kim HY, Lim CH, Lee YM, Moon YH. The occupational exposure level (OEL) for 2-bromopropane: the first OEL established by Korea. Appl Occup Environ Hyg 1999; 14: 356-358.
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