택사(Alismatis Rhizoma) 추출물의 급성 경구투여 독성 연구

석지현,노항식,정자영,하헌용,Seok, Ji-Hyun,Roh, Hang-Sik,Jeong, Ja-Young,Ha, Hun-Yong 대한한방안이비인후피부과학회 2013 한방안이비인후피부과학회지 Vol.26 No.4

Objectives : This study was carried out to evaluate the acute oral toxicity of Alismatis Rhizoma in Sprague-Dawley(SD) rats. Methods : male and female rats were administered orally with Alismatis Rhizoma water extract of 1,000 mg/kg (low dosage group), 2,000 mg/kg(middle dosage group) and 4,000 mg/kg(high dosage group). We daily observed number of deaths, clinical signs and gross findings for 7 days. After 7 days, we measured body and organs weight. Also we analyzed hematological changes. Results : No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology and serum biochemistry. In addition no significant changes of gross body and individual organs weight. Conclusions : These results suggest that water soluble extract of Alismatis Rhizoma has not acute oral toxicity and oral $LD_{50}$ value was over 4,000 mg/kg in SD rats.

백선피(Dictamnus dasycarpus Turcz) 추출물의 급성 경구투여 독성 연구

석지현,노항식,정자영,하헌용,Seok, Ji-Hyun,Roh, Hang-Sik,Jeong, Ja-Young,Ha, Hun-Yong 대한한방안이비인후피부과학회 2014 한방안이비인후피부과학회지 Vol.27 No.1

Objectives : This study was carried out to evaluate the acute oral toxicity of Dictamnus dasycarpus Turcz in Sprague-Dawley(SD) rats. Methods : Male and female rats were administered orally with Dictamnus dasycarpus Turcz water extract of 1,000 mg/kg(low dosage group), 2,000 mg/kg(middle dosage group) and 4,000 mg/kg(high dosage group). We daily observed number of deaths, clinical signs and gross findings for 7 days. After 7 days, we measured body and organs weight. Also we analyzed hematological changes. Results : No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology. But we found out subtle changes in body weight and individual organ weight of the female group. In addition specific changes were observed in serum biochemical value of female group. Conclusions : These results suggest that water soluble extract of Dictamnus dasycarpus Turcz has not acute oral toxicity and oral $LD_{50}$ value was over 4,000 mg/kg in SD rats. Also Dictamnus dasycarpus Turcz is expected to be sensitive with respect to the female.

임신랫드에 투여한 Butyl Benzyl Phthalate가 차산자에 미치는 영향

석지현(Ji Hyun Seok),황성희(Sung Hee Hwang),권은아(Eun A Kwon),김대용(Dae Yong Kim),김판기(Pan Gyi Kim) 한국환경보건학회 2000 한국환경보건학회지 Vol.26 No.1

Butyl benzyl phthalate(BBP) is a member of phthalic acid esters(PAEs) and is used extensively as a plasticizer in floor tiles, adhesives, synthetic leathers and hairspray. BBP is reported to be developmentally toxic, but it was to be nonregulated chemicals by Ministry of Environment in Korea. This study was investigated on the effect of BBP treatment on during gestational period of the dams and their fetuses. Sprague-Dawley rats were given BBP by oral administration at 0, 10, 100, 1000 mg/kg on days 1 to 21 of gestational period. One half of them was sacrificed at 20 days of gestation, and the rest was sacrificed at post parturition 21 days. Approximately one half of live fetuses in the gestational 20 days were randomly selected, fixed in 95% ethanol, stained alizarin red S and examined for skeletal malformations. At maternal findings, there were some significant changes in organ weight compared with control groups, especially in absolute liver weight and absolute kidneys weights by BBP but it was not dose dependent. As the BBP dose was increased; the uterus and ovary weight of treated dams were decreased in absolute and relative organ weight, except of 1000 mg/kg group. The post-implantation loss per litter was significantly increased at 1000 mg/kg treated group. As the BBP dose increased, the more dams showed estrus cycle changes such as estrus, proestrus earlier thaμ the control. In BBP 1000 mg/kg treated group, almost all pregnant rats had miscarried. At fetus findings, the body weights were decreased by administration of BBP. In female(F1) rats, there were significant changes in absolute and relative liver weight in BBP 10 mg/kg treated group. In genital organs, there were significant changes in absolute uterus weight in BBP 10 mg/kg treated group and there were inclined to increase of uterus weight in BBP 100 mg/kg treated group. In contrast, more high-treated group was inclined to decrease of ovary weight. In male(Fl) rats, there were significant changes in absolute and relative prostate weight in BBP 10 and 100 mg/kg treated group. Also, there were significant changes in absolute and relative testis weight in BBP 10 and 100 mg/kg treated group. The incidences of external malformations in fetuses were not examined. But skeletal variations were observed in all group. From these results, it could be concluded that the effects of pregngnt rats and their fetuses(F1) treated with BBP during day 1 to 20 of gestational period were estrogenic.

골담초 효모 발효 추출물의 피부 미백 효능에 관한 연구

석지현 ( Ji Hyun Seok ),이선영 ( Sun Young Lee ),채은정 ( Eun Jung Chae ),최신욱 ( Shin Wook Choi ) 대한화장품학회 2010 대한화장품학회지 Vol.36 No.3

본 연구는 미생물에 의한 골담초(Caragana sinica Rehder) 발효 추출물의 항산화 효과 및 피부 미백 효능에 관한 연구로 골담초에 S. cerevisiae KCTC 7913를 첨가하여 발효를 통해 얻어진 발효 추출물을 B16F10 멜라닌 세포에 처리한 결과, 농도의존적으로 멜라닌 생성 억제 효능이 증가한 것으로 확인하였으며, 추출물 내 유효 성분인 resveratrol의 함량이 발효 전의 추출물에 비해 증가함을 확인하였다. 멜라닌 생성 과정의 주요 단백질인 tyrosinase의 활성 억제 또한 골담초 추출물에 비해 골담초 발효 추출물의 효능이 우수함을 확인하였으며, 골담초 발효 추출물의 피부 자극 또한 일어나지 않는 것으로 확인되어 피부에 안전한 화장품 원료로서의 사용이 가능할 것으로 사료된다. In this study, we evaluated antioxidative effects and skin whitening effects of extract of Caragana sinica fermented by S. cerevisiae KCTC 7913. At first, Caragana sinica was fermented via inoculation of Saccharomyces cerevisiae KCTC 7913 and then extracted for fermented C. sinica. It has shown that more increased of melanogenesis inhibition activity in a dose-dependent manner on B16F10 melanoma cells and elevated the amount of resveratrol contents by HPLC analysis than non-fermented. Furthermore, the extract of fermented C. sinica was inhibitory effects against tyrosinase, a key enzyme of melanogenesis pathway, more than non-fermented C. sinica extract. And it did not show the skin irritation. Therefore, fermented C. sinica extracts might be used as safe cosmetic ingredients.

용담화 추출물의 미백 활성 연구

이현상 ( Hyun Sang Lee ),석지현 ( Ji Hyun Seok ),( Cheung Wai Ting ),김윤정 ( Yun Jeong Kim ) 대한화장품학회 2018 대한화장품학회지 Vol.44 No.1

본 연구에서는 항산화 및 미백 소재의 개발을 위해 다양한 추출법을 이용하여 용담화 추출물을 제조하고 항산화 효과 및 멜라닌 생합성 저해능을 평가하였다. 1,1-diphenyl-2-picrylhydrazyl (DPPH) 라디칼 소거활성을 평가한 결과 용담화 추출물의 항산화 활성은 모두 농도 의존적으로 증가하였다. 또한 용담화 추출물은 tyrosinase 활성을 저해시켰으며, B16F10 세포에서 멜라닌 생성을 감소시키는 효과를 보였다. 멜라닌 생합성에 관여하는 유전자의 발현에 미치는 영향을 알아보기 위해 tyrosinase-related protein (TRP)-1 및 2 mRNA 발현을 확인한 결과 용담화 추출물에 의해 TRP-1 및 2의 발현이 감소되는 것을 확인할 수 있었다. 이러한 결과로 보아 용담화 추출물은 항산화 및 미백 기능성 화장품 소재로서의 응용이 가능할 것으로 사료된다. In order to find new functional materials for the cosmetics application, we investigated the anti-oxidant and anti-melanogenic properties of Gentiana scabra extracts (GSE), which were prepared by the various extraction methods. Results showed that GSE had high DPPH radical scavenging activity in a dose-dependent manner. Also, GSE inhibited the prodution of melanin in B16F10 melanoma cell as well as tyrosinase activity. We also found that GSE inhibited mRNA expression of tyrosinase-related protein (TRP)-1 and 2. In conclusion we suggest that GSE is applicable to cosmetics as a potential ingredient for their anti-oxidant and whitening effects.

두날리엘라 살리나 추출물의 피부 열노화 억제 효과

주지혜 ( Ji-hye Joo ),석지현 ( Ji Hyun Seok ),홍인기 ( In-kee Hong ),김남경 ( Nam Kyoung Kim ),최은미 ( Eunmi Choi ) 대한화장품학회 2016 대한화장품학회지 Vol.42 No.1

자외선과 유사하게 열에 의한 콜라겐 분해와 비정상적인 탄력섬유의 축적을 증가시키는 현상을 열노화라 칭한다. 두날리엘라 살리나는 녹조류로 베타카로틴을 많이 함유하고 있어 건강식품으로 많이 이용되고 있으나 열에 의해 유도된 피부노화에서의 효능은 알려진 바 없다. 본 연구에서는 두날리엘라 살리나 에탄올 추출물의 항-열노화 효능을 확인하였다. 열을 가한 피부섬유아세포를 이용하여 MMP-1과 type I procollagen 발현을 ELISA를 이용해 확인하였다. 두날리엘라 살리나 추출물이 열에 의해 증가된 MMP-1 단백질 발현량을 감소시키며, type I procollagen 단백질 발현량은 증가시킨다는 사실을 확인하였다. 추가적으로, 두날리엘라 살리나 추출물에 의해 콜라겐 합성 과정에 관여하는 것으로 알려진 HSP47 mRNA의 발현이 증가함을 확인하였다. 또한, 두날리엘라 살리나 추출물이 염증매개인자(TGF-β, IL-12 등)의 발현을 감소시킴을 확인하였다. 다음으로 두날리엘라 살리나 추출물이 탄력섬유의 구성성분인 tropoelastin과 fibrillin-1 단백질 발현과 MMP-12 발현 조절을 통해 열에 의해 유도된 일광탄력섬유증을 조절하는 효능을 확인하였다. 이 결과를 통해 두날리엘라 살리나 추출물이 열에 의해 유도된 피부열노화를 효과적으로 예방함을 확인하였다. Just like UV radiation, heat increases collagen degradation and accumulation of abnormal elastin fiber and this is termed thermal skin aging. Dunaliella salina (DS), a green alga, is known for its beta-carotene accumulation, having various applications in the health and nutritional products. However, the effects of DS on heat-induced skin aging remain unexplored. In this study, we performed anti-thermal aging tests of the ethanol extract of DS (DSE). We measured the cellular levels of type I procollagen and MMP-1 using ELISA in human dermal fibroblast cells after heat shock. DSE reduced the expression of MMP-1 protein and increased the expression of type I procollagen. In addition, DSE upregulated the mRNA expression of HSP47 reduced by heat shock, which is involved in collagen synthesis. Also, DSE reduced the expression of inflammation mediator (TGF-β, IL-12, etc). We demonstrate that DSE regulates the heat-induced solar elastosis through the regulation of tropoelastin and fibrillin-1, two major proteins of elastic fibers, and MMP-12 expression. These results suggest that DSE may be effective for preventing thermally induced skin aging.

택사(Alismatis Rhizoma) 추출물의 반복 경구투여 독성 연구

노항식,석지현,정자영,이종권,김태성,최혜경,하헌용,Roh, Hang-Sik,Seok, Ji-Hyun,Jeong, Ja-Young,Lee, Jong-Kwon,Kim, Tae-Sung,Choi, Hye-Kyung,Ha, Hun-Yong 대한한방안이비인후피부과학회 2014 한방안이비인후피부과학회지 Vol.27 No.1

Objectives : This study was carried out to evaluate the repeated dose oral toxicity of Alismatis Rhizoma in Sprague-Dawley(SD) rats. Methods : Male and female rats were administered orally with Alismatis Rhizoma water extract of 500 mg/kg(low dosage group), 1,000 mg/kg(middle dosage group) and 2,000 mg/kg(high dosage group). We daily observed number of deaths, clinical signs and gross findings for 14 days(twice a day). After 14 days, we measured body and organs weight. Also we analyzed hematological changes. Results : No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology and serum biochemistry. In addition no significant changes of gross body and individual organs weight. Conclusions : These results suggest that water soluble extract of Alismatis Rhizoma has not repeated dose oral toxicity and oral LD50 value was over 2,000 mg/kg in SD rats. As a result, we can determine Alismatis Rhizoma is a relatively safe substance.

아데노바이러스 유전자치료벡터의 생식독성 연구

이규식,곽승준,김순선,이이다,석지현,채수영,정수연,김승희,이승훈,박귀례,Rhee, Gyu-Seek,Kwack, Seung-Jun,Kim, Soon-Sun,Lee, Rhee-Da,Seok, Ji-Hyun,Chae, Soo-Young,Chung, Soo-Youn,Kim, Seung-Hee,Lee, Seung-Hoon,Park, Kui-Lea 한국미생물학회 2007 미생물학회지 Vol.43 No.3

유전자치료W터의 주입시 생식세포를 통한 다음 세대로의 전달 가능성은 안전성 측면에서 관심을 중대시키고 있다. 특히 전립선암이나 난소암의 치료시 바이러스를 생식기관에 인접한 부위에 주입하여야 하므로 그 가능성이 높다. 따라서 본 연구에서는 유전자치료에 많이 이용되는 아데노바이러스를 매개로하여 tumor suppressor 유전자인 p53을 발현하는 아데노바이러스 벡터를 제조하여 이를 투여시 생식장기를 포함한 주요장기조직에의 분포와 germ cell을 통한 차세대로의 전달 가능성 등의 생식독성을 조사하였다. In vivo biodistribution study를 위하여 $Ad-CMV-{\beta}-gal$흑은 Ad-CMV-p53를 마우스 암 수의 복강에 주사한 후 생식장기를 포함한 주요 장기에서 아데노바이러스 유래 DNA검출 및 RNA발현 여부를PCR과 RT-PCR로 각각 확인하였다. 그 결과 간 및 비장과 같은 일반 장기에서도 주입한 외부유전자의 DNA가 검출되거나RNA가 발현되었을 뿐만 아니라, 정낭, 전립선, 부고환, 난소 및 자궁 등의 생식장기에서도 주입한 외부유전자가 검출되거나 발현되는 것으로 나타났다. Real-time PCR을 이용하여 각 장기에서의 투여된 아데노바이러스 벡터는 시간 의존적으로 감소되는 것을 정량하였다. Ad-CMV-p53를 암 수 마우스의 난소와 고환에 각각 직접 주사하여 교배시킨 후 그 후세대의 DNA를 분리하여 주입한 아데노바이러스 유래의 DNA를 검색한 결과, 어떠한 차세대에서도 주입한 아데노바이러스 유래의 DNA가 검출되지 않았다. 한편 생식장기에서의 PCR및 RT-PCR signal유래 vector의 위치를 확인하기 위해 매우 감도가 높은 in-situ PCR로 조사한 결과 고환의 경우 간질조직으로의 전달은 일어나나 정세관 내에는 아데노바이러스 벡터가 전달되지 않으며, 난소에서도 아데노바이러스벡터는 난포내의 난자에 전달되지 않고 기질조직에 존재하는 것으로 확인되었다. 결론적으로 복제 능력 이 결여된 아데노바이러스를 매개로 한 유전자치료제는 생식 장기에서 검출되더라도 다음 세대로 전달될 가능성은 대단히 낮음을 제시한다. The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

β-hexosaminidase 분비 억제 및 각질형성세포 분화에 대한 두충(Eucommia ulmoides Oliver) 추출물의 효과

홍인기 ( In Kee Hong ),김은지 ( Eun Ji Kim ),석지현 ( Ji Hyun Seok ),김보현 ( Bo Hyeon Kim ),장진동 ( Jin Dong Jang ),조기정 ( Gi Jung Joe ),최신욱 ( Shin Wook Choi ) 대한화장품학회 2014 대한화장품학회지 Vol.40 No.1

본 연구에서는 두충 추출물이 RBL-2H3 세포의 β-hexosaminidase의 분비 억제와 HaCaT keratinocytes 피부장벽의 회복과 관련한 filaggrin, transglutaminase-1 (TGase-1), cornified cell envelope (CE)의 발현에 미치는 영향에 관하여 연구하였다. β-hexosaminidase 방출 억제 능은 13% 효능을 확인하였고, 피부장벽기능의 회복과 관련된 인자들은 발현과 활성의 정도가 매우 우수한 것을 확인하였다. 각질형성세포의 분화를 판단할 수 있는 CE 측정에서는 두충추출물이 양성대조군보다 더 좋은 효능을 나타내기도 하였다. 따라서 두충 추출물은 β-hexosaminidase 분비 억제에 효과가 있으며, 손상된 피부장벽강화에 영향을 미치는 각질형성세포의 분화 촉진에 효과가 있음을 확인하였다. In this study, Eucommia ulmoides Oliver extracts was studied in order to see any effects on the β-hexosaminidase release suppression of RBL-2H3 cells and on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) related to the recovery of HaCaT keratinocyte skin barrier. Results showed that Eucommia ulmoides Oliver extracts reduced β-hexosaminidase release in RBL-2H3 cells and increased the effects of Eucommia ulmoides Oliver extract on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) in HaCaT keratinocytes. Taken together, these results suggested that Eucommia ulmoides Oliver extract may be applicable for keratinocyte differentiation.

원지(Root of Polygala teunifolia Willd)추출물의 급성 경구투여 독성 연구

노항식 ( Hang Sik Roh ),정자영 ( Ja Young Jeong ),석지현 ( Ji Hyun Seok ),하헌용 ( Hun Yong Ha ) 대한한방부인과학회 2013 大韓韓方婦人科學會誌 Vol.26 No.4

In this study, it was carried out to evaluate the acute oral toxicity of Root of Polygala teunifolia Willd. in Sprague-Dawley (SD) rats. Methods: Male and female rats were administered orally with Root of Polygala teunifolia Willd. water extract of 1,000 mg/kg (low dosage group), 2,000 mg/kg (middle dosage group) and 4,000 mg/kg (high dosage group). We daily observed number of deaths, clinical signs and gross findings for 7 days. After 7 days, we measured body and organs weight. Also we analyzed hematological changes. Results: No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology and serum biochemistry. But we found out histopathological changes in liver fat tissues of female. In addition, there were no significant changes of gross body and individual organs weight. Conclusions: These results suggest that water soluble extract of Root of Polygala teunifolia Willd. has not acute oral toxicity and oral LD50 value was over 4,000 mg/kg in SD rats.