• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1177-1182
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• 2012

Protein kinase C (PKC) has been implicated in carcinogenesis and displays variable expression profiles during cancer progression. Studies of dietary phytochemicals on cancer signalling pathway regulation have been conducted to search for potent signalling regulatory agents. The present study was designed to evaluate any suppressive effect of maslinic acid on PKC expression in human B-lymphoblastoid cells (Raji cells), and to identify the PKC isoforms expressed. Effects of maslinic acid on PKC activity were determined using a PepTag$^{(R)}$ assay for non-radioactive detection of PKC. The highest expression in Raji cells was obtained at 20 nM PMA induced for 6 hours. Suppressive effects of maslinic acid were compared with those of four PKC inhibitors (H-7, rottlerin, sphingosine, staurosporine) and two triterpenes (oleanolic acid and ursolic acid). The $IC_{50}$ values achieved for maslinic acid, staurosporine, H-7, sphingosine, rottlerin, ursolic acid and oleanolic acid were 11.52, 0.011, 0.767, 2.45, 5.46, 27.93 and $39.29\;{\mu}M$, respectively. Four PKC isoforms, PKC ${\beta}I$, ${\beta}II$, ${\delta}$, and ${\zeta}$, were identified in Raji cells via western blotting. Maslinic acid suppressed the expression of PKC ${\beta}I$, ${\delta}$, and ${\zeta}$ in a concentration-dependent manner. These preliminary results suggest promising suppressive effects of maslinic acid on PKC activity in Raji cells. Maslinic acid could be a potent cancer chemopreventive agent that may be involved in regulating many downstream signalling pathways that are activated through PKC receptors.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.116-120
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• 2004

The medicinal properties of Ganoderma lucidum have been recognized in China for many centuries. Active pharmaceutical components include triterpenes. To elucidate the molecular regulation of triterpene biosynthesis in this mushroom, a 57-base pair DNA fragment encoding the fourth conserved domain SQ-4 (SMGLFLQKTNIIRDYNEDL) of squalene synthase was synthesized and cloned into the expression vector pET-32a(＋). The recombinant fusion protein induced by IPTG (isopropyl-$\beta$-D-thiogalactopyranoside) was overexpressed in the Escherichia coli. Using the purified recombinant fusion protein of 20.9 kDa, a specific polyclonal antibody was obtained from immunized rabbit. Expression of squalene synthase at different development stages of Ganoderma lucidum was analyzed.

• Natural Product Sciences
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• 제21권3호
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• pp.176-184
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• 2015

In our search for natural soluble epoxide hydrolase (sEH) inhibitors from plants, an extract of the dried whole plants of Euphorbia supina Rafin was found to significantly inhibit sEH activity in vitro. Phytochemical investigation of E. supina resulted in isolation of 17 compounds (1 - 17), including triterpenes (1 - 4), phenolic compounds (5 - 8), and flavonoid derivatives (9 - 17). The structures of the isolated compounds were established mainly by extensive analysis of the 1D and 2D NMR, and MS data. All of the isolated compounds were evaluated for their sEH inhibitory activity. Among the isolated phenolic compounds, 8 was identified as a significant inhibitor of sEH, with an IC₅₀ value of 15.4 ± 1.3 μM. Additionally, a kinetic analysis of isolated compounds (2, 5, 8 - 11, 13, and 17) indicated that the inhibitory effects of flavonoid derivatives 10 and 11 were of mixed-type, with inhibitory constants (K_i) ranging from 3.6 ± 0.8 to 21.8 ± 1.0 μM, whereas compounds 2, 5, 8, 9, 13, and 17 were non-competitive inhibitors with inhibition K_i values ranging from 3.3 ± 0.2 to 39.5 ± 0.0 μM.

• Natural Product Sciences
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• 제22권3호
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• pp.154-161
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• 2016

Phytochemical investigation on the leaves of Pileostegia viburnoides Hook.f.et Thoms led to the isolation of twenty-five compounds, and their structures were identified as n-dotriacontane (1), taraxeryl acetate (2), friedelin (3), epifriedelinol (4), canophyllal (5), stigmast-4-en-3-one (6), stigmasterol (7), (24R)-5A-stigmastane-3,6-dione (8), ursolic acid (9), pomolic acid (10), umbelliferone (11), 4-epifriedelin (12), n-octatriacontanol (13), ${\beta}$-amyrin (14), ${\alpha}$-amyrin (15), taraxerol (16), nonadecanol (17), friedelane (18), arachic acid (19), protocatechuic acid (20), n-pentatriacontanol (21), hexadecanoic acid (22), vincosamide (23), daucosterol (24), and skimming (25), respectively. To our best knowledge, compounds 1, 2, 12, 13, 17 - 19 and 21-23 were new within Saxifragaceae family. Compounds 15, 16, and 20 were produced from this genus for the first time. Compounds 4, 14 and 25 were first obtained from species P. viburnoides and compounds 3, 5 - 11, and 24 were achieved from the leaves of P. viburnoides for the first time. Furthermore, the anti-neuroinflammatory activity of these isolates was evaluated.

• 한국약용작물학회지
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• 제24권6호
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• pp.437-443
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• 2016

Background: Previous phytochemical studies of whole Vernonia cinerea L. plants have identified sesquiterpene lactones, sterols, and triterpenes, which possess anticancer, antifeedant, and antimalarial activities. However, there are no reports of other types of bioactive metabolites. Therefore, the present study aimed to identify phenolic compounds with anti-inflammatory activity in the aerial parts of the plant. Methods and Results: Compounds were isolated from the aerial parts of V. cinerea using a silica and C-18 gel columns and semi-preparative HPLC instrument, and the structures of the compounds were determined using one- and two- dimension nuclear magnetic resonance spectroscopy and mass spectroscopy. The chloroform soluble extracts and isolated compounds were evaluated for their anti-inflammatory potential based on their ability to inhibit nitric oxide production and $TNF-{\alpha}$ induced $NF-{\kappa}B$ activity. Conclusions: Phytochemical study of the aerial parts of V. cinerea led to the isolation of six phenolic compounds. Compound 1 was a major metabolite, and to the best of our knowledge, compounds 2 - 6 were isolated from V.cinerea for the first time. Among the isolates, compounds 1 and 3 exhibited $TNF-{\alpha}$-induced $NF-{\kappa}B$ activity with $IC_{50}$ values of 7.5 and 11.5 M, respectively, and the inhibitory activity of phenyl propanoid compound 3 on $TNF-{\alpha}$-induced $NF-{\kappa}B$ was evaluated for the first time.

• Natural Product Sciences
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• 제22권1호
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• pp.60-63
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• 2016

The study is a pioneering effort to determine the mineral, nutritional, and phytochemical composition and phenolic content and to determine the free radical scavenging activity of Gigantochloa levis (Blanco) Merr, a native bamboo species (locally known as "bolo") in the Philippines. Proximate analysis showed that air-dried G. levis leaves contain 15.8% ash, 22.6% crude protein, 1.2% crude fat, 29.3% crude fiber, and 19.7% total sugar. Phytochemical tests indicated the presence of diterpenes, triterpenes, saponins, phenols, tannins, and flavonoids in both the ethanolic and aqueous leaf extracts, while phytosterols were only detected in the ethanolic extract. Folin-Ciocalteu assay determined the total phenolic content in gallic acid equivalents (GAE) to be $85.86{\pm}3.71$ and $32.32{\pm}1.01mg\;GAE/100g$ dried sample for the ethanolic and aqueous extracts, respectively. The total phenolic content in quercetin equivalents (QE) was $74.44{\pm}3.11$ and $29.43{\pm}0.85mg\;QE/100g$ dried sample for the ethanolic and aqueous extracts, respectively. The radical scavenging activity of the different solvent fractions containing varying concentrations of the extract was determined using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay. The ethyl acetate and 1-butanol fractions were found to have the highest radical scavenging activity. Mineral analysis via Energy Dispersive X-Ray Spectrometry (EDS) of the ash of G. levis leaves showed that Si is the major component, followed by K and Mg. These results point to the potential of G. levis leaves as a source of minerals and bioactive compounds with medicinal value.

• Mycobiology
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• 제36권3호
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• pp.173-177
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• 2008

The crude ethanol extracts (stem and fruits), their fractions and two triterpenes, $\beta$-Amyrin and 12-Oleanene 3$\beta$, 21$\beta$-diol, isolated as a mixture from the chloroform soluble fraction of an ethanolic extract of Duranta repens stem, were evaluated for antibacterial, antifungal activities by the disc diffusion method and cytotoxicity by brine shrimp lethality bioassay. The structures of the two compounds were confirmed by IR, $^1H$-NMR, $^{13}C$-NMR and LC-MS spectral data. The chloroform soluble fraction of stem and ethanol extract of fruits possess potent antishigellosis activity and also exhibited moderate activity against some pathogenic bacteria and fungi but the isolated compound 1 (mixture of $\beta$-Amyrin and 12-Oleanene 3$\beta$, 21$\beta$-diol) showed mild to moderate inhibitory activity to microbial growth. The minimum inhibitory concentrations (MICs) of the extracts (stem and fruits), their fractions and compound 1 were found to be in the range of 32$\sim$128 ${\mu}g/ml$. The chloroform soluble fractions of stem and ethanol extract of fruit showed significant cytotoxicity with $LC_{50}$ value of 0.94 ${\mu}g/ml$ and 0.49 ${\mu}g/ml$, respectively against brine shrimp larvae.

• 한국임상수의학회지
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• 제38권1호
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• pp.27-31
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• 2021

Shea butter (Vitellaria paradoxa) is a fat extracted from shea tree nuts and contains relatively high levels of non-glycerides. Triterpenes, the main non-glyceride component, exhibit a variety of biological activities such as antitumor, antibacterial, and anti-inflammatory. Shea butter extract (SBE) has been used to treat various skin problems such as burns, eczema, and rash in human medicine, but little is known about the activity of SBE on canine skin. This study evaluated the cytotoxicity and anti-inflammatory effect of SBE in canine keratinocytes. Cytotoxicity of lipopolysaccharide (LPS, 5-50 ng/mL) and SBE (50-200 ㎍/mL) was evaluated using the CCK-8 assay. Non-cytotoxic concentrations of LPS and SBE were administered to canine cell cultures to evaluate anti-inflammatory effects. To evaluate the anti-inflammatory activity of SBE, the levels of IL-1β, IL-8, IL-12, and TNF-α were measured using ELISA kits. The concentration of each cytokine was quantified in control, LPS-treated, LPS + SBE-treated groups. Increased levels of IL-1β, IL-8, and IL-12 were found in LPS-treated groups relative to control groups. LPS + SBE-treated groups showed a lower level of IL-1β, IL-8, and IL-12 than LPS-treated groups. These results suggest that SBE may have application as a topical agent for canine inflammatory skin diseases. However, further in vivo study is needed to evaluate the safety and efficacy of SBE in dogs.

• 한국식품과학회지
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• 제47권5호
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• pp.680-685
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• 2015

더덕 잎의 활용가치를 높이고자 더덕 잎 추출물 내 존재하는 기능성 생리활성 물질과 항산화 활성을 평가하였다. 먼저, 더덕 잎의 생리활성 물질의 존재를 검정하기 위해 파이토케미컬 검색을 한 결과, 더덕 잎 추출물에 환원당, 사포닌, 트라이터페노이드, 타닌 및 플라보노이드와 같은 생리활성 물질들이 존재함을 확인하였다. 이들 파이토케미컬의 존재를 근거로 하여 항산화 활성을 나타내는 것으로 알려진 총 폴리페놀 및 총 플라보노이드 함량을 측정하고, 이를 기반으로 더덕 잎의 항산화 활성을 평가하였다. 250 및 $500{\mu}g/mL$ 더덕 잎 추출물은 DPPH 라디칼 소거 실험에서 각각 43% 및 81%의 강력한 라디칼 소거 효과를 나타냈고, L-아스코브산의 71% 및 95%에 해당하는 환원력을 나타냈다. 또한, 더덕 잎 추출물 62.5, 125 및 $250{\mu}g/mL$에서 각각 73, 90 및 99%의 NO생성 저해활성을 나타내어 항산화 활성을 기반으로 한 항염증 효능의 가능성도 제공하였다. 이를 기반으로 더덕 잎 추출물의 아글리콘 분획을 조제하여 분석한 결과, 플라보노이드 화합물인 루테올린이 함유되어 있음을 확인하였다. 반면, 더덕과 같은 초롱꽃과(Campanulaceae)인 도라지의 항산화 연구에서 도라지 잎과 줄기 용매 분획물 중 줄기의 부탄올 분획물 $500{\mu}g/mL$을 첨가하였을 때 가장 높은 환원력과 화학성분을 보고한 바 있으나 항산화 활성 성분에 대해서는 언급하지 않았다(35). 루테올린이 함유되어 있는 더덕 잎 아글리콘 분획의 항산화 활성을 평가한 결과, 250 및 $500{\mu}g/mL$의 아글리콘 분획이 각각 67% 와 87%의 강력한 라디칼 소거 효과를 나타냈으며, L-아스코브산의 128% 및 141%에 해당하는 우수한 환원력을 나타냄을 확인하였다. 따라서, 우수한 항산화 활성 및 생리활성물질을 함유하고 있는 더덕 잎이 기능성 식품 소재로서의 활용가치가 높을 것으로 기대된다.

• Applied Biological Chemistry
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• 제49권4호
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• pp.270-275
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• 2006

Triterpenoid를 포함하고 있는 $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid를 애기마름으로부터 분리하였다. 이것은 pentacyclic triterpenes의 공통 구조를 가지며 amyrin ursolic acid 그룹에 속해 있다. 본 연구에서는 이 화합물의 독성 영향을 인간 간암 세포주인 HepG2에서 조사하였다. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid는 처리한 양에 비례하여 HepG2 세포주에서 독성을 보였다. 그리고 Confocal microscopy 결과는 $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid를 HepG2 세포에 처리한 시간에 비례하여 녹색 형광의 증가를 보여주었다. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid는 또한 HepG2 세포의 sub-G1 cell population 뿐만 아니라 DNA 분절(fragmentation) 현상의 증가를 보여 주었다. 이러한 결과는 $22{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid가 HepG2 세포에서 apoptosis를 통한 세포 사멸 유도를 의미한다.