체외 배양에 있어서 5% 산소 환경과 superoxide dismutase ( SOD ) 가 착상전 생쥐 배아 발달에 미치는 영향

양현원(Hyun Won Yang),이치형(Chi Hyeong Lee),오기석(Kie Suk Oh),유희석(Hee Sug Ryu),송승규(Seung Kyu Song),박동욱(Dong Wook Park) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.3

N/A Objective: In the human body the embryo initially gmws in the fallopian tube which is maintained in an 3-8% O2 concentration environment, and various substances such as growth factors and antioxidants present in tbe tubal fluid assists in maintaining a healthy environment for embryo development. But in IVF programs embryo cultures are conducted in incubators with 21.9% O2 and 5% CO2 condition, and such high oxygen concentrations have been reported to increase the production of oxygen free radicals within the embryo and is detrimental to the growth and development of the embryo. The objective of this study, therefore, is to determine the culture conditions which will decrease oxygen free radical production and thereby minimize the injury to the embryo. Methods: Six to eight week old ICR strain mice embryos were cultured in 5% or 21.9% O2 conditions and in culture media to which inaement concentrations of superoxide dismutase (SOD) had been and the H2O2 concentration within the embryo, embryo developmental rate, and degree of fragmentation of the embryos was investigated. Results: The control gmup embryos which were cultured in 21.9% O2 condition without addition of SOD showed developmental arrest at the 2-cell stage or fragmentation, while those cultured in 21.9% O2 condition with addition of SOD showed development to the blastocyst stage with deaeased fragmentation. In particular, the blastulation and fragmentation rates were the lowest in the group to which 500 IU/ml of SOD was added, but in the 5% O2 enviranment group many embryos reached the blastocyst stage and with no difference in frapnentation with or without addition of SOD. The HO relative intensity (120.5+-20.2) within the embryos cultured in 21.9% O2 environment without SOD was significantly higher than that (56.8+-10.8) of group with SOD (p<0.05). As showing that in the 5% O2 environment group without SOD it was 43.8+-7.8 and in the group with SOD it was 37.3+-5.4, the H2O2 concentration within embryos cultured in 5% 02 condition was significantly lower those that of 21,9% 02 environment regardless of SOD addition (p<0.05). Conclusion: The optimal oxygen concentration in incubator for mice embryo cultures is that which is similar to the 5% 0 concentration in vivo. When 20% 02 incubators are routinely used, the addition of SOD to the culture media will decrease the H2O2 concentration within the embryos with subsequent improvement in development. The optimal concentration which should be used is thought to be 500 IU/ml. It is suggested that the use of the above method in human IVF-ET programs will lead to improved embryo quality and enhanced pregnancy rates.

인간 자궁내막에서 Cyclooxygenase-1과 -2의 주기적 발현 양상

박동욱,양현원,권혁찬,황경주,유정현,이치형,김세광,조동제,오기석,Park, Dong-Wook,Yang, Hyun-Won,Kwon, Hyuek-Chan,Hwang, Kyung-Joo,Yoo, Jung-Hyun,Lee, Chi-Hyeong,Kim, Sei-Kwang,Cho, Dong-Jea,Oh, Kie-Suk 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1

Cyclooxygenase (COX) is an enzyme involved in the conversion of arachidonic acid to prostaglandins (PGs), and exists in two forms, COX-1 and COX-2. COX has been reported to be involved in early implantation by secretion of PGs which causes permeability of vessels and reaction of decidual cells around the implantation site. Recently, in mice and sheep studies, COX-1 and COX-2 expression in the endometrium has been reported to be different according to implantation and stages of the estrous cycle, but expression of COX-1 and COX-2 in human endometrium during the menstrual cycle has not yet been established. The purpose of this study was to observe the variances of COX-1 and COX-2 expression by immunohistochemical staining in endometrial samples obtained from human hysterectomy specimens and biopsies of women of reproductive age according to different stages of the menstrual cycle. Also, we attempted to observe COX-1 and COX-2 expression in the epithelial and stromal cells of the endometrium obtained during the mid-secretory phase, which were cultured separately. COX-2 showed a cyclic pattern of expression according to the different stages of the menstrual cycle and was strongly expressed particularly at the mid-secretory phase which corresponds to the time of implantation. However, COX-1 tended to be increased in the early proliferative, and mid- and late secretory phases, but was also expressed in the whole menstrual cycle showing no particular pattern. In the separately cultured cells COX-1 was expressed in epithilial cells and COX-2 in the stromal cells. The above results suggest that since COX-2 is expressed at the same time as implantation and cultured cells display a specific secretory pattern, COX-2 has inductive endocrine enzyme properties and has an important effect on endometrial cells during implantation. Also, COX-2 expression in endometrial cells may be utilized as a useful marker of endometrial maturation.

홍국발효홍삼의 이화학적 특성 및 생리활성

박진철(Jin Chul Park),차재영(Jae Young Cha),이치형(Lee Chi Hyeong),도은수(Eun Soo Doh),강인호(In Ho Kang),조영수(Young Su Cho) 한국생명과학회 2009 생명과학회지 Vol.19 No.11

홍국발효홍삼의 알코올성 간 질환 개선용 건강기능식품 개발을 위한 기초 자료를 얻기 위하여 일반 이화학적 특성과 항산화 활성 정도를 측정 하였다. 홍삼 및 홍국발효홍삼의 추출물 수율은 각각 수용성 5.83% 및 6.58%, 70% 에탄올 0.98% 및 0.62%, 70% 메탄올 3.04% 및 1.27%로 수용성 추출에서 가장 높았다. 홍삼 및 홍국발효홍삼 1% 추출물 중의 총 당도는 각각 0.23% 및 0.17%로 홍삼에서 약간 높았으며, 가용성 고형분도 각각 0.6 및 0.5 Brix로 홍삼에서 약간 높았다. 총 폴리페놀 화합물 함량은 홍삼보다 홍국발효홍삼 추출물에서 높았으며, 70% 에탄올 추출물에서 가장 높았다. 미네랄 함량은 K가 22,386 및 16,936 ppm으로 가장 많이 함유되어 있었고, 다음으로 Ca이 각각 3,693 및 2,310 ppm으로 함유되어 있었고, Mg는 각각 2,647 및 2,703 ppm, Na는 각각 103 및 109 ppm, Fe는 각각 115 및 42 ppm, Zn은 각각 57 및 62.3 ppm이 함유되어 있었다. 항산화 실험에서 총 폴리페놀 함량, DPPH radical 소거능 및 불포화 지방산 linoleic acid를 이용한 TBA법에 의한 항산화 활성은 홍삼 추출물 보다는 홍국발효홍삼에서 높은 것으로 나타났다. This study investigated the biological activities and chemical characteristics of Monascus-fermented Korean red ginseng (MFRG). The comparative activities of water, ethanol, and methanol extracts from MFRGE and Korean red ginseng (RG) were tested in vitro of antioxidative models of linoleic acid peroxidation by thiocyanate and thiobarbituric acid (TBA) methods and DPPH (α,α'-diphenyl-β-picrylhydrazyl) radical scavenging activities. In addition, measurements of their bioactive total phenolic compounds and minerals, and extract yield, were obtained. The extract yield of each solvent extracted from MFRG and RG was aqueous by 6.58% and 5.83%, ethanol by 0.62% and 0.98%, and methanol by 1.27% and 3.04%, respectively. Total phenolic compounds were higher in all solvents extracted from MFRG than those from RG. Major mineral contents (ppm) of MFRG and RG were K by 16,936 and 22,386, Ca by 2,310 and 3,693, Mg by 2,703 and 2,647, respectively. The DPPH radical scavenging activities were higher in all solvents extracted from MFRG than those from RG, however, all these extracts exhibited a relatively low level of radical-scavenging activity compared to the butylated hydroxytoluene (BHT). In antioxidative activities determined by TBA method using linoleic acid peroxidation, 70% methanol extract from MFRG and RG showed the highest antioxidative activity at a concentration of 0.1%. These results may provide the basic data to understand the biological activities of bio-active materials derived from MFRG.

생쥐 배아의 체외배양시 저농도 산소 환경이 배아 발달에 미치는 영향

유정현(Jong Hyun Woo),황경주(Kyung Joo Hwang),양현원(Hyun Won Yang),이치형(Chi Hyeong Lee),양정인(Jeong In Yang),권혁찬(Hyuck Chan Kwan),오기석(Kie Suk Oh) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.12

N/A Objective: It is known that mouse embryos before implantation develop in a low oxygen environment of 3- 8% concentration and with antioxidant materials such as vitamins, antioxidant enzymes, ferrous binding proteins, and albumin in follicular and tubal fluids. However, the 20% oxygen culture condition with chemically defined media might be produce an abundance of ROS, and leads to developmental delay or developmental block in vitro. In this study, we attempt to elucidate the relationship between intracellular H2O2 production and embryo development in different oxygen culture conditions of mouse embryos. Methods: Prenuclear embryos from C57BL/CBA Fl hybrid and ICR mouse were cultured in incubators which provided 5% carbon dioxide, 20% oxygen and 5% carbon dioxide, 5% oxygen. Measurement of H2O2 level in a embryo was performed with DCHFDA(2,7 -dichlorodihydroflourescein diacetate)and analyzed with Quanti-cell 700, and the number of blastomeres was counted with DAPI( 4, 6'-diamidino-2-phenylindole). Results: Oxygen concentration of the culture medias was significantly higher in the 20% oxygen environment compared to that of 5% oxygen environment. Culture of mice embryos in high oxygen condition leads to high HO concentrations at 2 cell stage and developmental delay or 2-cell block regardless of the strain. But in a 5% oxygen environment, which is similar to in-vivo conditions HO production was suppressed continuously through out culture and development of embryos was definitely improved. Conclusion: These results suggest that there is a difference in the production of ROS or protective mechanism according to the mouse strains and stage of development, and it is thought that in-vitro culture in 5% oxygen environment provides stable in vivo equilibrium but in a 20% oxygen environment there is production of ROS which overcome the protective mechanism which leads to cellular damage and embryo developmental delay.

알코올 급여 흰쥐의 혈청 지질 및 조직 과산화지질 농도에 미치는 홍국발효홍삼분말의 영향

차재영(Jae Young Cha),안희영(Hee Young Ahn),엄경은(Kyung Eun Eom),박보경(Bo Kyung Park),전방실(Bang Sil Jun),박진철(Jin Chul Park),이치형(Chi Hyeong Lee),조영수(Young Su Cho) 한국생명과학회 2009 생명과학회지 Vol.19 No.7

홍국균을 홍삼 분말에 접종하여 발효시킨 홍국발효홍삼을 이용하여 알코올 투여 흰쥐의 혈중 지질 농도 및 조직 내 산화스트레스에 미치는 영향에 대하여 검토하였다. 혈중 총 지질 및 유리지방산 농도는 알코올 대조군(AC)에서 증가가 홍국발효홍삼 투여군(AMFRG)에서 감소경향을 보였으며, 중성지질 농도는 양성 대조군(AS) 및 홍삼 투여군(ARG)에서 유의적으로 감소하였다. 혈청 총 콜레스테롤 농도는 정상 대조군(NC)에 비해 알코올 대조군(AC)에서 유의적으로 증가하였고, 홍국 발효홍삼 투여군(AMFRG)에서 알코올 대조군(AC) 보다는 낮았지만 통계상의 유의적인 차이는 없었다. 혈중 HDL-cholesterol 농도는 정상 대조군(NC)에 비해 알코올 투여군 모두에서 증가하는 경향을 보였는데, 특히 홍국발효홍삼 투여군(AMFRG)에서 유의적으로 증가하였다. 동맥경화지수(AI)는 정상 대조군(NC)에 비해 알코올 대조군(AC)에서 증가한 반면 양성 대조군(AS) 및 홍삼 투여군(ARG)에서 감소경향을 보였고, 홍국발효홍삼 투여군(AMFRG)에서 유의적으로 감소하였다. 간장, 심장, 비장, 고환에서 생체막 과산화지질 생성 정도는 정상 대조군(NC)에 비해 알코올 대조군(AC)에서 유의적으로 증가하였고, 이러한 증가는 홍국발효홍삼 투여군(AMFRG)에서 유의적으로 감소하였다. 과산화지질 생성 촉진작용이 있는 비헴철 함량도 알코올 대조군(AC)의 간 조직에서 증가한 반면 홍국발효홍삼 투여군(AMFRG)에서 유의적으로 감소하였고, 간 조직 내 내인성 항산화 물질인 glutathione 농도는 알코올 대조군(AC)에서 유의적으로 감소하였고, 홍국발효홍삼 투여군(AMFRG)에서 유의적으로 증가하여 상호간의 높은 상관관계를 나타내었다. 이상의 실험결과 홍국발효홍삼은 홍삼과 홍국의 상호작용에 의한 시너지 효과로 알코올 투여 동물실험에서 혈중 지질 개선효과는 물론 조직 내 항산화 활성증가로 산화스트레스를 경감시키는 효능이 있는 것으로 사료되었다. The effects of Monascus-fermented Korean red ginseng (MFRG) on the contents of serum lipids and tissues lipid peroxidation was investigated in alcohol feeding rats (AC group). Serum contents of total lipid and free fatty acid in alcohol feeding rats were significantly increased, but these increases tended to decrease in the AMFRG group. Serum triglyceride content was also significantly decreased in the AMFRG group compared to other groups. Serum content of total-cholesterol was significantly increased in AC group compared to normal control (NC) group, whereas there was no significant difference between the AC and AMFRG groups. Content of HDL-cholesterol in serum was slightly increased in the AC group compared to the NC group, but this increase in the AC group was more significantly increased in the AMFRG group. At the same time, atherogenic index (AI) was also significantly decreased in the AMFRG group compared to the AC group. Contents of thiobarbituric acid reactive substances (TBARS) in the liver, heart, spleen and testes were significantly increased in the AC group compared to the NC group, but these increases were significantly less in the AMFRG group. Contents of liver nonheme ion was increased in the AC group and was significantly decreased in the AMFRG group, which suggested that lipid peroxidation contents are inversely correlated with liver nonheme ion content. Hepatic glutathione concentration was significantly decreased in the AC group, but this content was significantly increased in the AMFRG group and it showed the antioxidant abilities of glutathione. These results suggested that Monascus-fermented Korea red ginseng has anti-atherogenic index (AI) effects as well as antioxidative activities through reduced tissue oxidative stress in alcohol feeding rats.

홍국발효홍삼 분말 급여에 의한 흰쥐의 혈청 지질 농도 및 항산화 활성에 미치는 영향

차재영(Jae Young Cha),박진철(Jin Chul Park),안희영(Hee Young Ahn),엄경은(Kyung Eun Eom),박보경(Bo Kyung Park),전방실(Bang Sil Jun),이치형(Chi Hyeong Lee),조영수(Young Su Cho) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.9

Monascus purpureus KCCM 12002 홍국균으로 발효시킨 홍국발효홍삼 분말을 식이 중에 1% 수준으로 첨가하여 4주간 흰쥐에 급여한 후 혈청 임상생화학적 특성과 조직 내항산화 활성 및 간 조직에 미치는 영향을 검토하였다. 체중 증가량, 식이섭취량 및 음료섭취량은 각 실험 군 간에 유의적인 차이는 없었다. 간 조직 총 중량 및 상대 중량은 다른 실험 군에 비해 홍국발효홍삼 분말 투여 군에서만 유의적으로 증가하였다. 간 손상 임상지표인 AST 및 γ-GTP 활성은 대조군에 비해 홍국발효홍삼 분말 투여 군에서 현저하게 저하하였으나, ALT 활성에는 영향을 미치지 못하였다. 혈중 총 지질 및 중성지질 농도는 대조군에 비해 silymarin 투여 군에서 유의적으로 감소하였고, 홍삼 분말 투여 군에서는 감소경향을 보인 반면 홍국발효홍삼 분말 투여 군에서는 유의적인 증가를 나타내었다. 혈청 총 콜레스테롤 농도는 대조군에 비해 silymarin, 홍삼 분말 및 홍국발효홍삼 분말투여 군들에서 모두 저하경향을 보였다. 혈중 HDL-콜레스테롤 농도는 대조군에 비해 홍국발효홍삼 분말 투여 군에서만 유의적으로 증가하였다. 따라서 총 콜레스테롤 농도에 대한 HDL-콜레스테롤 농도의 비로 나타내는 동맥경화지수(AI)는 대조군에 비해 Silymarin 및 홍삼 분말 투여 군에서 유의적으로 감소하였고, 홍국발효홍삼 분말 투여 군에서도 현저하게 감소하였다. 간 조직의 과산화지질(TBARS)을 측정한 결과, 대조군에 비해 홍국발효홍삼 분말 투여 군에서는 유의적으로 감소하였으나, silymarin 및 홍삼 분말 투여 군에서는 감소경향을 보였다. 특히 홍국발효홍삼은 간 조직내에서 높은 과산화지질 억제작용을 나타내어 조직 내 항산화 물질로 잘 알려진 glutathione 농도는 대조군에 비해 silymarin 및 홍국발효홍삼 분말 투여 군에서 유의적으로 증가하였으며, 홍삼 분말 투여 군에서만 증가경향을 보였다. 간 조직의 조직학적 검사를 실시한 결과 대조군에서는 혈관을 중심으로 간 소엽 구조가 잘 유지되었으며, 간세포들은 풍부한 호산성 세포질과 둥근 핵을 가지고 있었고 간세포판은 대부분 한 층으로 잘 유지되어 있었으며, silymarin, 홍삼분말 및 홍국발효홍삼 분말 투여 군에서도 대조군과 같은 현상을 보였다. 이상의 실험결과 홍국발효홍삼은 항동맥경화 작용뿐만 아니라 간 조직 내 항산화물질 증가에 의한 산화스트레스를 경감시킴으로써 향후 간 독성 개선 효능을 가지는 건강기능식품 개발의 기초 소재로 활용될 가능성이 높은 것으로 판단되어진다. Monascus-fermented Korean red ginseng (MFRG), fermented by Monascus purpureus KCCM 12002, may be an ideal candidate for the hepatoprotectic, hypolipidemic and antioxidative activities. Effect of MFRG powder on these parameters in rats was investigated. Body weight gain, food intake, and water consumption were not significantly different among the groups. Total and relative weights of liver were significantly higher in MFRG group than that in other groups. The activities of AST and γ-GTP were highly lowered in MFRG group compared to control group. Contents of serum total lipid and triglyceride were significantly lowered in silymarin group and were significantly increased in MFRG group compared to control group, but tended to be lowered in RG group. Serum content of total cholesterol tended to be lowered in silymarin, RG, and MFRG groups compared to control group. HDL-cholesterol contents was only significantly increased in MFRG group compared to control group. At the same time, atherogenic index (AI) was also significantly lowered in silymarin, RG and MFRG groups compared to control group, and this effect was more pronounced in MFRG group. Content of thiobarbituric acid reactive substances (TBARS) in the liver was significantly lowered in MFRG group and tended to lowered in silymarin and RG groups compared to control group. The hepatic glutathione concentration was significantly higher in silymarin and MFRG groups. Hepatic morphology in all experimental groups revealed clear-cut hepatic lobules with the uniform pattern of the polyhedral hepatocytes radiating towards the periphery from the central vein. These results suggested that MFRG may have anti-atherogenic index (AI) and antioxidative activity in normal dietary feeding rats.

임신과 동반된 중증 근무력증 1례

주연종,이치형,양정인,김행수,오기석,박문성 大韓周産醫學會 1997 Perinatology Vol.8 No.1