Phytochemical Constituents from the Rhizomes of Osmunda japonica Thunb and Their Anti-oxidant Activity

우경완,정자균,이현주,김태묵,김민석,정호경,안병관,함성호,전병훈,조현우 한국생약학회 2017 Natural Product Sciences Vol.23 No.3

Eleven compounds (1-11) were isolated from the rhizomes of Osmunda japonica, and their structures were elucidated based on 1H, 13C-NMR and LC-IT-TOF MS data. Of these compounds, all compounds (1 - 11) have been previously reported, although five (6 - 9, 11) have not previously been isolated from this plant. The antioxidant activities of isolated compounds (1 - 11) were measured by DPPH and ABTS assays, and compound 10 showed the high antioxidant activity.

한라돌쩌귀로부터 분리된 Dopaol β-D-glucoside의 신장독성 보호효과

노종현,정자균,정호경,장지훈,정다은,이기호,김아현,성태경,박호,조현우 한국약용작물학회 2017 한국약용작물학회지 Vol.25 No.4

Background: Cisplatin is one of the most extensively used chemotherapeutic agents for the treatment of cancer, including bladder, and ovarian cancers. However, it has been shown to induce nephrotoxicity, despite being an outstanding anti-cancer drug. In this study, we investigated the protective effect of dopaol β-D-glucoside (dopaol) on cisplatin-induced nephrotoxicity. Methods and Results: To confirm the protective effect of dopaol on cisplatin-induced nephrotoxicity, HK-2 cells were treated with 20 μM cisplatin and 80 μM dopaol. Cisplatin increased apoptosis, caspase-3 activity and mitochondrial dysfunction; however pretreatment with 80 μM dopaol successfully attenuated apoptosis, caspase-3 activity and mitochondrial dysfunction. To evaluate the protective effect dopaol on cisplatin-induced nephrotoxicity in vivo, we used an animal model (balb/c mice, 20 ㎎/㎏, i.p. once/day for 3 day). The results were similar to those obtained using HK-2 cells; renal tubular damage and neutrophilia induced by cisplatin reduced following dopaol injection (10 ㎎/㎏, i.p. once/day for 3 day). Conclusions: These results indicate that dopaol treatment reduced cisplatin-induced nephrotoxicity in vitro and in vivo, and can be used to treat cisplatin-induced nephrotoxicity. However, further studies are required to determine the toxicity high dose dopaol and the signal pathways involved in its mechanism of action in animal models.

노랑붓꽃에서 분리된 Iridin의 독소루비신 유도 HK-2 세포 괴사에 대한역할 및 암세포에 대한 작용

노종현,이기호,정호경,이무진,장지훈,심미옥,정자균,정다은,조현우 한국자원식물학회 2018 한국자원식물학회지 Vol.31 No.2

Doxorubicin is a anti-cancer drugs that interferes with the growth and spread of cancer cells in human body. Doxorubicin is used to treat different types of cancers that affect the ovary, thyoid and lungs, but induced side effect such as nephrotoxicity and cardiotoxicity. Thus, we investigated that the effect of iridin on doxorubicin-induced necrosis in HK-2 cells, a human proximal tubule cell. To confirm effect of iridin on doxorubicin-induced necrosis, HK-2 cells are treated with 10 μM doxorubicin and 80 μM iridin. 80 μM iridin reduced 10 μM doxorubicin-induced necrosis, the mitochondrial over activation and caspase-3 activation. However, iridin reduces anti-cancer effect of doxorubicin such as PARP1 and caspase-3 activation, checkpoint proteins (CDK4 and CDK6) in NCI-H1129 cells (Human non-small cell lung cancer cell). In HCT-116 cells (Human colorectan cancer cell), iridin do not increased protein expression of CDK4 and CDK6 decreased by doxorubicin. Results indicate that treatment of iridin was diminished doxorubicin-induced necrosis in HK-2 cells. However, iridin was decreased anti-cancer effect of doxorubicin on NCI-H1229, but not HCT-116. Thus, further experiment are required to iridin treatment on various cancer cells and animal models because effect of iridin different cell type. Key words - Doxorubicin, HK-2, Iridin, Iris koreana Nakai, Necrosis, Nephrotoxicity 노랑붓꽃에서 분리된 iridin의 doxorubicin으로 유도된 신장세포괴사 모델에 대한 보호 효과 및 암세포에 대한 작용을 알아보기위해 연구를 수행하였다. Iridin 단일 처리로는 신장근위세뇨관 세포주에 대해 독성을 나타내지 않았으며, 80 μM의 농도에서 10 μM doxorubicin 처리에 의한 세포사멸을 94.6 ± 2.6% 까지 회복시켰다. 또한 80 μM iridin 처리는 10 μM doxorubicin 처리에 의해 증가된 cleaved PARP1과 cleaved caspase-3를 포함하는 세포사멸 신호전달을 차단하였을 뿐만 아니라 DNA fragmentation, necrotic cell death 및 mitochondrial dysfunction을개선시켰다. 마지막으로 암세포에서 iridin의 효과를 확인해본 결과, 폐암세포주인 NCI-H1229 세포에서 doxorubicin의 항암효과를 억제하는 경향이 나타났지만 대장암 세포주인HCT-116 세포주에서는 암세포에 대한 성장억제를 방해하지 않는 것으로 확인되었다. 따라서 폐암세포에서 doxorubicin과iridin의 병용처리는 힘들다고 판단되고, In vivo 수준에서 신장독성 및 대장암 관련 실험을 통해 iridin의 역할을 추가적으로확인해야한다고 생각된다.

Loperamide로 유도된 변비모델에서 Lactobacillus casei에 의해 발효된 볶은 결명자 물 추출물의 효과

노종현,정호경,이무진,장지훈,심미옥,정자균,이기호,안병관,조정희,장민철,용주현,조현우 한국약용작물학회 2016 한국약용작물학회지 Vol.24 No.6

Background: Constipation is one of the most common functional gastrointestinal disorder. The present study examined the ability of water extract of fermented (FRC) and non-fermented (NFRC) roasted Cassia tora to improve intestinal function and reduce constipation in a rat constipation model.Methods and Results:Different concentration of FRC and NFRC were orally administered loperamide (5 ㎎/㎏; LOP) reduced the number, weight, and water content of feces, as well as intestinal transit motility. However, 24 h-(24 hour fermented roasted-Cassia tora) 300 ㎎/㎏ FRC administration increased the number, weight, and water concent of feces, compared to that seen in the LOP group, and also improve intestinal transit mitility and, the thickness of distal colon and mucous fluid.Conclusions:The results of the present study indicated that LOP-induced constipation was improved by treatment with FRC. Therefore FRC could be used to develop functional foods or natural medicine for constipation. However, further study is needed to clarify how fermentation improves the medicinal properties of roasted C. tora.

조팝나무 뿌리 열수 추출물이 RAW264.7 세포에서 미치는 항산화 및 항염증 활성

심미옥,이현주,장지훈,이효은,정호경,김태묵,노종현,정자균,정다은,조현우 한국자원식물학회 2017 한국자원식물학회지 Vol.30 No.4

본 연구에서는 LC IT TOF MS를 이용하여 조팝나무 뿌리의열수 추출물과 메탄올 추출물을 분석하였다. 그 결과, 열수 추출물에서 caffeic acid와 p-coumaric acid가 주성분으로 검출되었으나, 메탄올 추출물에서는 열수 추출물에서 검출되지 않은 저극성 물질들이 다수 검출되었다. 조팝나무 메탄올 추출물의 항산화 및 항염증에 대한 연구는 보고된바가 있으나, 열수추출물에 대한 연구는 아직 미비한 실정이다. 따라서, 본 연구에서는 조팝나무 뿌리 열수 추출물(SSN)의 항산화활성 및 항염증활성을 탐색하고자 하였다. 먼저 항염증 활성을 탐색하기 위해lipopolysaccharide (LPS)에 의해 활성화된 대식세포로부터 분비되는 NO함량을 측정하였다. SSN은 LPS로 유도한 염증반응에서 세포 독성 없이 NO의 생성을 농도 의존적으로 억제하였다. 다음으로, 항산화활성을 측정하기 위해 DPPH, ABTS 라디칼 소거능과 SOD 유사활성을 측정한 결과 SSN은 강한 유리라디칼저해능을 나타냈으며, 이는 SSN이 폴리페놀(56.7 ㎎/g)과 플라보노이드(15.1 ㎎/g)와 같은 생리활성 물질을 함유하고 있어 강한 항산화능을 가지는 것으로 사료된다. 또한, 대식세포에서H2O2로 유도한 세포독성을 완화시켰으며, 세포내 ROS 생성을억제함에 따라 천연물 유래 항산화제로서의 가치를 확인하였다. 이상의 결과를 종합해 볼 때 SSN이 항산화와 항염증 효과와관련된 건강보조식품 및 기능성화장품 소재로의 활용이 가능할것으로 사료된다. Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai (SSN) has been used for the anti-inflammation in traditional folk medicine. To compare water and methanol extracts of SSN, we analyzed major components using LC IT TOF MS. The major components of hot water extract were identified as caffeic acid and p-coumaric acid, but methanol extract was not well established. However, methanol extract was detected with less polarity compounds compared to hot water extract. Next, we investigated the inhibitory effects of SSN water extract on the lipopolysaccharide (LPS)-induced inflammatory response or H2O2-induced oxidative stress in Raw 264.7 macrophage cells. SSN strongly suppressed the production of nitric oxide in LPS-induced inflammatory response without cytotoxcity. The SSN possessed free radical scavenging activities such as DPPH (IC50=320.2 ㎍/㎖), ABTS (IC50=124.0 ㎍/㎖), and superoxide anion radical (IC50=122.6 ㎍/㎖). The total phenol and flavonoid content of SSN was 56.7 ㎎/g, and 15.1 ㎎/g, respectively. Furthermore, SSN decreased the H2O2-induced cytotoxicity by enhancing the cell viability, and SSN significantly reduced the intracellular reactive oxygen species (ROS) level. Therefore, SSN may be recommended as an effective strategy to prevent and/or treat various inflammation and ROS-induced diseases.

인동덩굴로부터 분리된 Cynaroside이 Doxorubicin으로 유도된 인간 근위세뇨관 HK-2 세포의 괴사에 미치는 저해 효과

노종현,정호경,이무진,장지훈,심미옥,정자균,정다은,안병관,조현우,Nho, Jong Hyun,Jung, Ho Kyung,Lee, Mu Jin,Jang, Ji Hun,Sim, Mi Ok,Jung, Ja Kyun,Jung, Da Eun,An, Byeong Kwan,Cho, Hyun Woo 한국약용작물학회 2017 한국약용작물학회지 Vol.25 No.5

Background: Cynaroside is a flavone, a flavonoid-like compound, known by different names (luteoloside and cinaroside). It is commonly found in Lonicera japonica Thunb., Chrysanthemum moriflium, and Angelica keiskei. The process of cell death has been classified as necrosis and apoptosis. Necrosis refers to unregulated cell death induced by a chemotherapeutic agent. Doxorubicin is an anthracycline anti-cancer drug used to treat acute leukemia, cancer, and lymphoma. However, it induces nephrotoxicity including tubular damage. Therefore, we investigated the protective effect of cynaroside against doxorubicin-induced necrosis in HK-2 cells. Methods and Results: To confirm the beneficial effect of cynaroside on doxorubicin-induced necrosis, HK-2 cells, a human proximal tubule epithelial cell line were treated with $10{\mu}M$ doxorubicin and $80{\mu}M$ cynaroside. Doxorubicin treatment resulted in increased DNA fragmentation, caspase-3 activity and mitochondria hyperactivation during cell necrosis. However, pretreatment with $80{\mu}M$ cynaroside attenuated DNA fragmentation, caspase-3 activity and mitochondria hyperactivation induced by $10{\mu}M$ doxorubicin in HK-2 cells. Conclusions: These results indicated that pretreatment with cynaroside ameliorated doxorubicin-induced necrosis in HK-2 cells. Therefore, cynaroside be used as a therapeutic agent for improving doxorubicin-induced nephrotoxicity. However, further studies are required to evaluated the toxicity of cynaroside treatment in animals and to determine its protective effect against doxorubicin-induced nephrotoxicity in an animal model.

선복화 에탄올 추출물의 Nitric Oxide 생성, 산화스트레스 및 대장암 세포 억제효과

노종현,정다은,정호경,이무진,장지훈,심미옥,정자균,조현우 한국약용작물학회 2018 한국약용작물학회지 Vol.26 No.1

Background: Inula japonica Thunb. is a plant belonging to the family compositae. Inulae flos (flower of I. britannica var. chinensis Regal.) is the dried flower of I. japonica Thunb. and contains various flavonoids (patulitrin, nepitrin and kaempferol), which have been utilized in traditional oriental medicine to treat nausea, phlegm, and coughs. However, ethanol extract of I. britannica (IJE) has not been previously studied for its use in cancer treatment, and its effects on oxidative stress, or inflammation. Thus, the present study investigated the anti-oxidant, anti-inflammatory, and anti-colorectal cancer effects of IJE using RAW264.7 and HCT- 116 cells, which are human colorectal cancer cell line. Methods and Results: IJE contained flavonoids (80.95 ± 5.3 ㎎/g) and polyphenols (310.53 ± 10.6 ㎎/g). Moreover, it reduced lipopolysaccharide (LPS)-induced nitric oxide (NO) production and H2O2-induced oxidative stress by decreasing reactive oxygen species (ROS) levels. Additionally, the 500 ㎍/㎖ IJE treatment increased caspase-3 activity and apoptotic cell death in HCT-116 cells. Conclusions: These results demonstrate that the anti-cancer effect of IJE against human colorectal cancer cells involves caspase-3 activation and apoptotic cell death. IJE also inhibited LPS-induced NO production, and H2O2-induced oxidative stress in RAW264.7 cells. However, further studies are required to explore how IJE treatment regulates signal transduction in NO and ROS production.