임신 중 환경오염물질 2-Bromopropane에 노출된 마우스의 모독성 평가

허정두(Jeong-Doo Heo),김충용(Chung-Yong Kim),강성철(Chen-Zhe Ziang),서정은(Jeong-Eun Suh),정문구(Moon-Koo Chung),김무강(Moo-Kang Kim),신동호(Dong-Ho Shin),김종춘(Jong-Choon Kim) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1

2-Bromopropane (2-BP), a halogenated propane analogue, is a substitute for chlorofluorocarbones (CFCs). The present study was carried out to investigate the potential adverse effects of 2-BP on pregnant dams after maternal exposure during the gestational days (GD) 6 through 17 in ICR mice. The test chemical was administered subcutaneously to pregnant mice at dose levels of 0, 500, 1000, and 1500 ㎎/㎏/day. All pregnant females were subjected to autopsy on GD 18. Treatment-related clinical signs, as evidenced by dose-dependent increases in the incidence and severity of rough fur, swelling, induration, crust formation, and ulceration at the injection sites, were observed at dose levels of above 1000 ㎎/㎏/day. Maternal body weight on GD 18 and maternal body weight gain for the intervals GD 6-17 (treatment period) were suppressed at a dose level of 1500 ㎎/㎏ in a dose-dependent manner, but not statistically significant. Hematological investigations revealed a dose-dependent decrease in the number of platelet at a dose level of 1500 ㎎/㎏. Fetal body weights of both sexes in the 1500 ㎎/㎏ group were also decreased in a dose-dependent manner. There were no adverse effects on mortality, food consumption, organ weights, and reproductive findings except for decreased fetal body weight in any treatment group. Based on these results, it was concluded that the 12-day repeated subcutaneous dose of 2-BP resulted in an increased incidence of clinical signs, suppressed maternal body weight, reduced platelet count and decreased fetal body weight at a dose level of 1500 ㎎/㎏/day in pregnant mice. In the present experimental conditions, the no-observed-adverse-effect level of 2-BP was considered to be 500 ㎎/㎏ for dams.

Cynomolgus Monkey의 발란티디움 감염증 사례

한수철(Su-Cheol Han),허정두(Jeong-Doo Heo),하창수(Chang-Su Ha),신호철(Ho-Chul Shin),김충용(Choong-Yong Kim) 한국실험동물학회 2003 Laboratory Animal Research Vol.19 No.3

The comparative toxicities of BPA, BPS, BPF, and TMBPF on the development, healthspan and aging

Moonjung Hyun(현문정),Sung min Han(한성민),Jeong-Doo Heo(허정두) 환경독성보건학회 2021 한국독성학회 심포지움 및 학술발표회 Vol.2021 No.5

Bisphenol A (BPA) is a widely used substance in daily life and industry. BPA is well known for its adverse health effects. Several alternative materials have been developed to replace it. Here, we comprehensively analyze the efficacy of BPA and three substitutes, including bisphenol S (BPS), bisphenol F (BPF), and tetramethyl bisphenol F (TMBPF), on organismal aging, healthspan, and energy homeostasis using an in vivo Caenorhabditis elegans (C. elegans) model animal and two cultured mammalian cell lines. All bisphenols resulted in decreased reproductive ability and lifespan and increased axonal beading, a sign of neuronal aging. BPA resulted in abnormalities in development and growth, such as slow growth rate, short body length, and disorganized vulva tissue morphology. Each substitute also showed detrimental effects on at least two or more of these events. Exposure to all bisphenols except TMBPF, proposed as a relatively safe BPA alternative, caused mitochondrial abnormalities such as decreased oxygen consumption and mitochondria membrane potential. In contrast, organismal ATP level was markedly increased by all bisphenols, including TMBPF. In mammalian cells, BPA increased ATP level and decreased ROS but did not affect mitochondrial permeability transition pore (mPTP). Only TMBPF exhibited a significant increase in ROS and mPTP opening. Notably, all of the bisphenols caused a severe decrease in cell viability. Studies using these in vivo animals and cell cultures suggest that the potentially harmful physiological effects of other BPA alternatives, including TMBPF, compared to BPA should be considered.

Embryo lethality and teratogenicity of 2-Bromopropane in the Sprague-Dawley rat

김종춘,오기석,신동호,김성호,김현영,윤효인,강성철,허정두,정문구,Kim, Jong-Choon,Oh, Ki-Seok,Shin, Dong-Ho,Kim, Sung-Ho,Kim, Hyeon-Yeong,Yun, Hyo-In,Jiang, Cheng-Zhe,Heo, Jeong-Doo,Chung, Moon-Koo The Korean Society of Veterinary Science 2003 大韓獸醫學會誌 Vol.43 No.4

The present study was undertaken to evaluate the potential adverse effects of 2-BP on pregnant dams and embryo-fetal development after maternal exposure during the gestational days (GD) 6 through 19 in Sprague-Dawley rats. The test chemical was administered subcutaneously to pregnant rats at dose levels of 0, 375, 750 and 1250 mg/kg/day. During the test period, clinical signs, mortality, body weights and food consumption were examined. All dams were subjected to caesarean section on GD 20 and their fetuses were examined for external, visceral and skeletal abnormalities. At above 750 mg/kg, toxic effects including signs of toxicity, suppressed body weight, decreased gravid uterine weight and reduced food intake were observed in pregnant dams. An increase in the fetal deaths, a decrease in the litter size, a reduction in the fetal body weight and an increase in the incidence of fetal morphological alterations were also found. There were no adverse effects on either pregnant dams or embryo-fetal development at a dose level of 375 mg/kg. These results suggest that a 14-day subcutaneous dose of 2-BP is embryolethal and teratogenic at above 750 mg/kg/day in pregnant rats. In the present experimental condition, the no-observed-adverse-effect level of 2-BP is considered to be 375 mg/kg/day for dams and embryo-fetuses, respectively.

Cynomolgus Monkey에 있어 혈액학 및 혈액생화학적 지표

김충용(Choong-Yong Kim),이현숙(Hyun-Sook Lee),한수철(Su-Cheol Han),허정두(Jeong-Doo Heo),하창수(Chang-Su Ha),권명상(Myung-Sang Kwon),정문구(Moon-Koo Chung) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1

To provide basic information on blood for safety assessment, physiological levels of hematology and serum chemistry were ascertained in nineteen male (average body weight, 2.86 ㎏) and sixteen female (average body weight, 2.73 ㎏) cynomolgus monkeys. Hematologic parameters included fourteen items including red blood cells (RBC) hemoglobin (Hb), hematocrit (HCT), white blood cells (WBC) and differential leukocyte counts (%WBC). Serum biochemical parameters included twenty items including alkaline phosphatase (ALP), creatinine (CREA), creatine phosphokinase (CPK), triglyceride (TG), inorganic phosphorous (IP), etc. Hematological levels showed great variation in platelet, reticulocyte and WBC, whereas showed less variation in RBC, Hb, and HCT. Serum biochemical levels showed great variation in aspartate aminotransferase (AST), alanine aminotransferase (ALT), ALP, Blood urea nitrogen (BUN), glucose (GLU), total cholesterol (TCHO), CPK, TG, total bilirubin (TBIL), phospholipid (PL) and inorganic phosphate (IP), whereas showed less variation in albumin/globulin ratio (A/G ratio), total protein (TP), albumin (ALB), sodium (Na), potassium (K), chloride (Cl). Hematological determined showed greater values of Hb, and HCT in males than in females, while serum biochemical assay showed greater values of ALP, CPK and IP in males than in females. The present study will be valuable in the proper interpretation and validation of general toxicology studies using cynomolgus monkeys.

인체유래 폐세포주에서 염화 벤잘코늄에 의한 독성 영향

김혜진(Hyejin Kim),현문정(Moonjung Hyun),이선민(Seonmin Lee),허정두(Jeong-Doo Heo) 환경독성보건학회 2021 한국독성학회 심포지움 및 학술발표회 Vol.2021 No.5

염화 벤잘코늄은 어린이들 상처의 살균 소독제로 수십년 동안 안전하게 사용되어왔다. 하지만 폐로 들어갈 경우 지극히 높은 독성을 보이며 2시간 이내에 사망한다고 알려져 있다. 최근, 코비드-19 pandemic으로 인해 손소독제 및 손세정제 사용이 증가하고 있는데 여전히 국내에서 시판되는 손세정제 1200여종 중 123종에서 염화 벤잘코늄이 함유되어 있다. 따라서, 손소독제 및 손세정제 사용이 증가하고 일상생활에 노출되고 있어 독성을 유발할 가능성이 농후하기 때문에 안전성 평가가 필요하다. 인체유래 폐세포주를 활용하여 세포 생존율과 미토콘드리아를 기반으로 하는 활성산소 및 막투과성을 확인하였다. 염화 벤잘코늄 혼합물 농도 의존적으로 세포 생존율이 감소하였으며, 활성산소는 증가하고 막투과성이 감소하는 현상을 확인하였다. 또한, 활성산소와 막투과성에 관련된다고 알려진 미토콘드리아 STAT3 단백질이 염화 벤잘코늄을 포함하는 손세정제와 염화 벤잘코늄 혼합물 노출에 의해 감소하는 현상을 확인하였다. 또한 STAT3를 knockdown 하였을 때 대조군에 비해 세포 생존율이 감소하며 활성산소가 증가하는 현상을 확인하였다. 본 연구를 통해 염화 벤잘코늄 노출에 대한 독성 영향을 미토콘드리아 STAT3 단백질 수준에서 평가할 수 있는 방법을 제시하고자 한다.

Eritadenine 함유 표고버섯균사체 고체배양 효소추출물(EA-LEMSCEE)의 spontaneous hypertension rat 혈압감소

문연규(Yeon-Kyu Moon),정재은(Jae Eun Jung),조복임(Bok-Im Cho),허정두(Jeong-Doo Heo),최정일(Jungil Choi),김정옥(Jeong Ok Kim),하영래(Yeong Lae Ha) 한국생명과학회 2018 생명과학회지 Vol.28 No.2

표고버섯으로부터 유래한 eritadenine (EA)은 혈중 low density lipoprotein (LDL), triglyceride (TG), phospholipid를 감소시키는 물질이다. 최근 저희 연구팀은 in vitro 연구에서 EA가 angiotensin converting enzyme (ACE)의 활성을 저해함을 밝혔다. 본 연구에서는 EA와 EA 함유한 표고버섯균사체 고체배양 효소추출물(EA-LEMCEE)이 본태성 고혈압 쥐 SHR의 혈압을 낮추었음을 보고한다. EA 및 EA-LEMSCEE는 인체 탯줄 혈관내피세포 EA.hy926 세포의 ACE 활성을 억제하였다. EA-LEMSCEE는 SHR의 수축기 및 이완기 혈압을 Control 처리구에 비해 유의성 있게 낮추어 처리 5주에는 Positive control인 captopril (CP) 처리와 비슷한 혈압감소 효과를 나타내었다. EA-LEMCE는 LEMSCEE에 비해 수축기 및 이완기 혈압을 감소시켰지만 유의성은 없었다. EA- LEMSCEE는 renin과 ACE 활성을 Control 처리에 비해 유의성 있게 억제하였고, angiotensin II (Ang II) 함량을 감소시켜, 처리 5주 후에 이러한 CP의 효과와 유사하였다. 또한 EA-LEMCEE는 LEMCEE에 비해 유의성 있게 renin 및 ACE 활성을 억제하였고 및 Ang II 함량을 감소시켰다. 이와 같은 결과는 EA 및 EA-LEMCEE는 본태성 고혈압 SHR의 renin과 ACE 활성을 억제하여 혈압을 낮추었고, EA 및 EA-LEMSCEE는 인체 고혈압을 낮출 수 있음을 의미한다. Eritadenine (EA), derived from Lentinus edodes (LE), reduced low-density lipoprotein (LDL), triglyceride (TG), and phospholipids in bloods, and fatty acid depositions in animals and humans. Previously, we reported that EA inhibited angiotensin-converting enzyme (ACE) activity in vitro. Now, we report that EA reduced blood pressures in spontaneous hypertension rats (SHR). EA-containing LE mycelial culture enzyme extract (EA-LEMSCEE) was prepared from LE mycelial solid cultures and the hot-water extract of LE fruit bodies. Both EA and EA-LEMSCEE inhibited ACE activity in immortalized human umbilical endothelial cells (EA.hy926). EA-LEMSCEE treatments (7.5 mg/kg, 22.5 mg/kg) significantly reduced systolic and diastolic blood pressure in SHR. At five weeks of treatment, EA-LEMSCEE treatment significantly reduced systolic and diastolic blood pressure, similar to the positive control (captopril, CP; 4 mg/kg) treatment. In addition, the LEMSCEE without EA decreased systolic and diastolic blood pressures compared to the control, but not significant. EA-LEMSCEE decreased renin and ACE activities, and angiotensin II (Ang II) contents in SHR compared to the control. After five weeks of treatment, the effect of EA-LEMCEE was similar to that of CP. These results indicate that EA and EA-LEMSCEE reduce blood pressure by inhibiting the renin and ACE activity of SHR. Furthermore, these results imply that EA or EA-LEMSCEE could be used as an antihypertension agent in humans.

RWPE-1 전립선세포에서 eritadenine을 함유한 신령버섯균사체 액체배양물의 항염증효과 및 항산화효과에 의한 전립선비대증 관련 biochemical marker 개선 효과

하영래(Yeong Lae Ha),문연규(Yun-Gu Moon),김나현(Na-Hyun Kim),허정두(Jeong Doo Heo),조민정(Min Jung Cho),김예라(Ye Ra Kim),김영숙(Young Suk Kim),김정옥(Jeong OK Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.10

신령버섯균사체 액체배양 추출물(Agaricus blazei mycelial liquid culture extract, ABMLCE)은 TM-3 mouse Leydig 고환세포에서 testosterone (TS)의 생성을 촉진하였다. 이번 연구에서는 eritadenine (EA, 15.3 ㎎/100 g)을 함유한 ABMLCE가 RWPE-1 전립선세포에서 전립선비대증(Benign prostatic hyperplasia, BPH) 관련 핵심 효소인 5α-reductase 2 (5α-R2) 활성과 dihydrotestosterone (DHT) 함량을 감소시켰음을 보고한다. RWPE-1 전립선 세포는 24-well plate에서 ABMLCE (0~50 ppm), EA (0~10 ppm,), finasteride (FS 10 μM: positive Control)를 함유한 Keratinocyte serum-free medium (K-SFM)에서 24시간 배양하였다. 시료처리 세포배양액에 대해 5α-R2, superoxide dismutase (SOD), catalase (CAT), cyclooxygenase-2 (COX-2) 효소활성, TS, DHT, tumor necrosis factor-α(TNF-α)와 interleukin-1β (IL-1β)를 assay kit를 사용하여 분석하였다. 5α-R2 활성과 DHT 함량은 ABMLCE의 농도 의존적으로 유의성(p<0.05) 있게 감소되었고, SOD와 CAT 효소활성은 유의성(p<0.05) 있게 증가되었다. 반면에 COX-2 활성과 TNF-α 및 IL-1β의 함량은 감소되었다(p<0.05). 마찬가지로 EA의 효과도 ABMLCE 효과와 유사하였다. ABMLCE 50 ppm과 EA 10 ppm의 5α-R2 및 DHT 감소효과는 FS 10 μM 효과와 유사하였다. 이들 결과는 ABMLCE와 EA는 항염증효과와 항산화효과를 통하여 BPH 유발 핵심 인자인 5α-R2 효소활성과 DHT 함량을 감소시켰다. 또한 이 결과는 EA를 함유하는 ABMLCE는 인체 BPH 개선 소재로 활용할 수 있음을 의미한다. Agaricus blazei mycelial liquid culture extract (ABMLCE) promoted the production of testosterone (TS) in TM-3 mouse Leydig testis cells. Now, we report that ABMLCE containing eritadenine (EA) as a minor constituent (15.3 mg/100 g) reduced 5α-reductase 2 (5α-R2) enzyme activity and dihydrotestosterone (DHT) content which are key constituents for the benign prostatic hyperplasia (BPH) inductions. RWPE-1 prostate cells were grown in a Keratinocyte serum-free medium (K-SFM) containing ABMLCE (0~50 ppm), EA (0~10 ppm,), and finasteride (FS 10 μM: a positive control) in a 24-well plate for 24 hr. Supernatants collected from cell-cultured media were used for the assay of 5α-R2, superoxide dismutase (SOD), catalase (CAT) and cyclooxygenase-2 (COX-2) enzyme activities, and for TS, DHT, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) contents by their assay kits. The 5α-R2 activity and DHT content were proportionally reduced (p<0.05) to concentrations of ABMLCE. The SOD and CAT enzyme activities were significantly (p<0.05) elevated concomitant with ABMLCE concentrations, while COX-2, TNF-α and IL-1β showed reverse results (p<0.05). Similarly, the effects of EA were similar to those of ABMLCE. Efficacies of ABMLCE 50 ppm and EA 10 ppm in 5α-R2 and DHT reduction were similar to those of 10 μM FS. These results suggest that ABMLCE and EA reduced 5α-R2 and DHT through their anti-inflammatory and anti-oxidative actions. This implies that ABMLCE containing EA could be a beneficial material in the cure of BPH in humans.

연구논문 : 실험용 게잡이 원숭이의 pair-housing 효과 2. 혈액학적 지표의 변화

김지은 ( Jee Eun Kim ),한수철 ( Su Cheol Han ),이필수 ( Pill Soo Lee ),허정두 ( Jeong Doo Heo ),김충용 ( Choong Yong Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.2

The effect of the social enrichment program using mingle cages was investigated in hematological parameters of male cynomolgus monkeys. It is generally believed that the social enrichment program using mingle cages may offer the socialization and the prevention of isolation syndrome such as abnormal behavior. The total of 48 males and 42 females were housed in pairs homosexually with mingle cages reformed from individual cages for 9 weeks and we observed the clinical signs, body weight changes and hematological parameters. In the previous study, Han et al.,2007 observed clinical signs and body weight changes. 93.8 % of male cage mates and 100% of female cage mates were made successful pair-housing formation except 3 cases of male cage mates with serious injuries from fighting. And body weight gains were not changed between individual cage-housing and pair-housing for 9 weeks. However WBC count of male monkeys in individual cage was decreased compared with pair housing group after 8 and 21 weeks from pair housing. Significant increase in neutrophils and decrease in lymphocytes were observed in pair housing group. Thus, this finding indicate that housing condition may change hematological parameters. The present finding may serve as useful basic data for social enrichment program using mingle cage.

실험용 게잡이 원숭이의 pair-housing 효과 2. 혈액학적 지표의 변화

김지은 ( Jee Eun Kim ),한수철 ( Su Cheol Han ),이필수 ( Pill Soo Lee ),허정두 ( Jeong Doo Heo ),김충용 ( Choong Yong Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.2

The effect of the social enrichment program using mingle cages was investigated in hematological parameters of male cynomolgus monkeys. It is generally believed that the social enrichment program using mingle cages may offer the socialization and the prevention of isolation syndrome such as abnormal behavior. The total of 48 males and 42 females were housed in pairs homosexually with mingle cages reformed from individual cages for 9 weeks and we observed the clinical signs, body weight changes and hematological parameters. In the previous study, Han et al.,2007 observed clinical signs and body weight changes. 93.8 % of male cage mates and 100% of female cage mates were made successful pair-housing formation except 3 cases of male cage mates with serious injuries from fighting. And body weight gains were not changed between individual cage-housing and pair-housing for 9 weeks. However WBC count of male monkeys in individual cage was decreased compared with pair housing group after 8 and 21 weeks from pair housing. Significant increase in neutrophils and decrease in lymphocytes were observed in pair housing group. Thus, this finding indicate that housing condition may change hematological parameters. The present finding may serve as useful basic data for social enrichment program using mingle cage.