Background : The 1,2-unsaturated PAs, reported to be widely present in medicinal plants belonging to Asteraceae, Boraginaceae, and Fabaceae, cause hepatotoxicity and genotoxicity in humans and animals. Hence, there is a need for an analytical method that allows these dangerous plant toxins to be determined. In this study, we developed a method that can be used for the rapid and accurate determination of nine toxic PAs in medicinal plants using ultra-pressure liquid chromatography–electrospray ionization–quadrupole–time-of-flight mass spectrometry (UPLC-ESI-Q-TOF). Methods and Results : The compounds were eluted onto a C18 column with 0.1% formic acid and acetonitrile, and separated with good resolution within 11 min. all analytes was characterized by its precursor ions generated by ESI-Q-TOF and fragment ions produced by collision-induced dissociation (CID), which were used as a reliable database. The proposed analytical method was verified with reference to the ICH guidelines. The proposed UPLC-ESI-Q-TOF method was applied to four medicinal plants, and lycopsamin, echimidine, senkirkine and senecionine were detected by matching with reference standard, and additional six PAs were tentatively identified though chemical profiling. In addition, the QuEchERS method was the most efficient in comparison with methods like hot water and methanol in extraction efficiency of pyrrolizidine alkaloids. Conclusion : The our proposed method can determine PAs rapidly and accurately in medicinal plants and will be utilize as an important data for other researchers who need analytical information of PAs.

Background : Turmeric (Curcuma longa L.) and black pepper (Piper nigrum L.) have traditionally been used as Asian medicinal and culinary herb. Curcumin, a major compound of turmeric, has been known to have antitumor activity. However, curcumin is bioavailable because it is rapidly metabolized and released from the body. Therefore, the addition of adjuvants such as piperine, a potent inhibitor of drug metabolism, is one of the ways to increase the bioavailability of curcumin. Methods and Results : The yields of turmeric and black pepper ethanolic extracts (TM and BP) are 18.2 and 8.2% (w/w), respectively. The EC50 values of A549 and NCI-H292 cells exposed to TM were 77.8 ㎍/㎖ and 92.0 ㎍/㎖, respectively. No significant cytotoxicity was observed up to the 400 ㎍/㎖ in the A549 and NCI-H292 exposed to BP. Based on the central composite design, the co-treatment of TM and BP enhanced the cytotoxicity of A549 and NCI-H292 cells. The optimal combination concentration (optimal EC50 value) of TM and BP calculated by the response surface methodology assay were 48.5 and 241.7 ㎍/㎖. The conbination index assay confirmed that the cytotoxic effect at optimal combinatino concentration was due to the synergistic effect. Conclusion : We hypothesized that co-treatment of TM and BP enhanced cytotoxicity more than single treatment of TM against lung cancer cells, and cell death at this time may synergetic cytotoxicity effects associated with curcumin metabolism.

Background : The effects of heat treatments on the phycochemical charateristics and antioxidant activities of ginkgo nuts were investigated by roasting at various temperatures (150, 180, and 210℃) for 60 min. Methods and Results : For each roasting temperatures, pH, total acidity, chromaticity, total cyanide content, reducing power, total polyphenol content, 2,2’-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities were compared and analyzed. As the roasting temperature and time increased, the pH and total cyanide content decreased from 7.32 to 6.31 and from 697 to 282 ㎍/㎎, respectively, while total acidity increased from 0.40 to 0.84%. There was an 11 or more color difference (ΔE) between the non-heated ginkgo nut and heated ginkgo nuts, indicating a remarkable color difference. The antioxidant activities, including reducing power, total polyphenol content, and DPPH and ABTS radical scavenging activities increased rapidly at 210℃ according to increasing in roasting temperature and time. Conclusion : These results suggest that the ginkgo nut with heat treatment could be used in food materials and medicines for decreasing cyanide content as well as for increasing antioxidant compound content.

Background : F. fomentarius extracts have been recently reported to process immune-stimulatory and anti-inflammatory effects. In this study, we evaluated the anti-mutagenic effect of F. fomentarius ethanol extracts and the effective chemical components from the extract were analyzed by LC-Q-TOF. Methods and Results : F. fomentarius was extracted with 70% ethanol yielding a crude extract 6.8%. The crude extract was fractionated sequentially to diethyl ether, chloroform, dichloromethane, butanol and water with a yield of 43.9%, 2.958%, 6.8%, 21.6% and 25.5.%, respectively. The anti-mutagenic effect of F. fomentarius extract and its fractions was tested in Ames test by two type of Salmonella typhimurium (TA98 and TA100). Among the five fractions, diethyl ether has shown the highest and significant anti-mutagenic effect (98.3%, at 3,000 ㎍/plate concentration). Moreover, we investigated the chemical constituents of the extract using UPLC-Q-TOF. A total of 10 compounds such as flavonoids (velutin, 3, 4′,5-Trihydroxy-3′,7-dimethoxyflavanone) and fatty acids (γ-linoleic acid, stearic acid) and deterpenoid (kirenol) were tentatively identified with an accurate mass within 10 ppm mass error. Also, flavonoids and fatty acids were detected with higher rate than other compounds based on obtained chromatograms. Conclusion : We demonstrated that F. fomentarius extract and fractions have anti-mutagenic activity through Ames test. Furthermore, we will carry out isolation of each components from its fraction and use them to conduct additional anti-mutagenic test.

Background : Insulin-like growth factor 1 (IGF-1) appears to enhance the differentiation of osteoblasts and to activate the mineralization of bone. Hence, the aim of this study was to investigate the effects of ginseng complex on the remodeling of rats tibia. Methods and Results : Ginseng complex significantly increased serum IGF-1 by 58% and 34.5% than the control, respectively. Treatment with α-amylase when manufacturing these extracts remarkably increased the concentration of IGF-1 by 63% and 36% above the control, respectively. This ways that this ginseng complex, especially α-amylase treated extracts, contained a higher level of IGF-1 secretion in the ginseng complex groups. In addition, increases of 8% in femuf length were found after 12 weeks of oral administration with ginseng complex (300mg/kg). Conclusion : These results mean that ginseng complex have beneficial effects on bone effects on bone growth via IGF-1.

Background : Recently, many studies to seek for medicinal herb compounds from natural products have been attempted capable of being free from harmful side effects and reducing the economic burden. This study was conducted to investigate whether Morus alba L.(MAL) water extracts has a biological safety, and an inhibitory potential against the mutagenicity induced by cigarette smoke condensates(CSC). Methods and Results : MAL was extracted with 70% ethanol and the yield of the extract was 35.1%. The 70% ethanol MAL extract was fractionated sequentially by diethylether, chloroform, dichloromethane, and water, respectively. Crude MAL 70% ethanol extract itself and its solvent fractions did not show any mutagenic effect up to 1 mg/plate toward Salmonella typhimurium TA 98 with or without the microsomal S-9 mixture of liver metabolic activations. On the other hand, the crude MAL extract showed an inhibitory activity against the mutagenicity of CSC with S-9 mixture. Diethyl ether layer among five solvent fractions showed the highest inhibitory activity at the same dose. The inhibitory activity of diethyl ether layer was also increased dose-dependently and the inhibitory rate was about 97.1% at the concentration of 1 mg/plate. Conclusion : In this study, we concluded that all of the crude MAL 70% ethanol extract and its solvent layers are potentially safe for mutagenicity and the diethyl ether layer among the crude MAL 70% ethanol extract has an inhibitory effect against the mutagenicity of CSC.

Background : Korean Red Ginseng has been reported to have various anti-virus, anti-tumor, anti-inflammation and immune-regulating activity. However, Korean Red Ginseng Crude Saponin (KRGCS) effect on Pulmonary inflammation has been insufficient research. We have investigated the effect of KRGCS on pulmonary inflammation in mouse model. Methods and Results : Pulmonary inflammation induced by cigarette smoke condensates (CSC) and lipopolysaccharide (LPS) in BALB/c mice. 100uL (ratio 1:1) of CSC (4mg/mL) and LPS (100ug/mL) solution instilled with intratracheal injection once per a week for 3 weeks in mice. KRGCS was also administered orally a dose of 10mg/kg and 25mg/kg for 3 weeks. KRGCS treatment groups decreased the number of inflammatory total cells in lung tissues and bronchoalveolar lavage fluid (BALF) and the number of neutrophils in BALF dropped more than negative control groups. Additionally, KRGCS reduced the absolute number of CD4+ and CD8+ cells in BALF, and the number of CD11b+ Gr-1+cells in the lung and BALF. Furthermore, we found that KRGCS declined significantly the level of the cytokine chemokine (C-X-C motif)-1 (CXCL-1), Macrophage inflammatory protein-2 (MIP-2), and tumor necrosis factor alpha (TNF-α) in BALF. Conclusion : These results that KRGCS has an inhibitory effects on pulmonary inflammation such as COPD, may be a candidate of effective herbal pharmaceuticals for the cure of lung diseases.

In this study, essential oils were extracted from the leaf of Chamaecyparis obtusa (CLEO), indigenous toKorea, CLEO constituents were analysed, and the effects of CLEO on airway hyperresponsiveness (AHR) and airwayinflammation (AI) were investigated in Ovalbumin (OVA)-induced asthma mouse model. Terpenoid components amongidentified CLEO constituents made up more than 80%. The CLEO-treated group in comparison to the control groupshowed reduced AHR, the decrease of eosinophil number in the bronchoalveolar lavage fluid (BALF), reduced specific anti-OVA IgE level in the serum, and a significant reduction in Th2 cytokines levels in the BALF with concentration. We con-cluded that CLEO have an alleviating effect on asthma-like symptoms such as AHR and AI. Further studies about antiasth-matic effect are necessary on the focus of single component of CLEO.

Traditionally, Ligustrum lucidum fruits (LL) is one of the well-known oriental herb used in the treatment of skin and lung inflammation. This study investigated anti-inflammatory effects of LL in the pathogenesis of acute pulmonary inflammation in mice. Acute pulmonary inflammation was induced by intratracheal instillation of cigarette smoke condensate (CSC) and lipopolysaccharide (LPS) 5 times within 12 days in mice. LL extract was administered orally at a dose of 50 or 200 mg/kg. Administration of LPS and CSC significantly elevated airway hyperresponsiveness (AHR) to mice, and increased in the levels of inflammatory cells and mediators in mice. However, the LL extract significantly reduced the elevated AHR, and the increase of neutrophils, CD4+/CD3+ cells and CD8+/CD3+ cells, along with reducing the expression of TNF-α, IL-6, and MIP-2. Moreover, the LL extract alleviated the infiltration of inflammatory cells in expanded airway walls histologically. These results indicate that the LL extract has an inhibitory effects on acute pulmonary inflammation and AHR in murine model, and plays a crucial role as a immunomodulator which possess anti-inflammatory property.

The feature of asthma are airway inflammation (AI), reversible airway obstruction, and an increased sensitivity to bronchoconstricting agents, elevated airway hyperresponsiveness (AHR), excess production of Th2 cytokines, and eosinophil accumulation in the lungs. This study was performed to investigate if oral administration of Scutellaria baicalensis Georgi water extracts (SBG) have the antiasthmatic potential for the treatment of asthma. Asthmatic HI and AHR were induced by systemic sensitization to ovalbumin (OVA) with intratracheal instillation with 0.1 mg/mL of diesel exhaust particles (DEP) suspension once a week for 10 weeks in BALB/c mice. SBG was orally administered with the concentraion of 200 mg/kg 5 days a week for 10 weeks. Long-term SBG treatment suppressed the eosinophil infiltration into airways from blood, the asthmatic AI and AHR by attenuating the production of cytokine IL-4, IL-5 and IL-13, histamine and OVA-specific IgE. Our data suggest that SBG has inhibitory effects on AI and AHR in a mouse model of asthma, may act as a potential Th2 cytokine antagonist, and may have a therapeutic effect on allergic asthma.

This study was carried out to investigate the inhibitory effect of Angelicae Dahuricae Radix (ADR) extract on chronic obstructive pulmonary disease (COPD) induced by cigarette smoke condensate (CSC) and lipopolysaccharide (LPS) in mice. COPD was induced by intratracheal instillation of LPS and CSC 5 times for 12 days; this increased airway hyperresoponsiveness (AHR) and inflammatory cells in bronchoalveolar lavage fluid (BALF). ADR extract was administered orally at a dose of 50 and 200 mg/kg. The concentration of imperatorin, a major component of ADR and therefore used as a measure of quality control, was 0.098%±0.018%. Treatment of the mice with ADR extract (50 and 200 mg/kg) alleviated AHR and reduced inflammatory cell counts. Treatment with cyclosporin A (CSA; 10 mg/kg) also modulated AHR and reduced inflammatory cells effectively. Compared with CSA treatment, treatment with ADR (50 mg/kg) extract reduced neutrophil and CD4+/CD3+ cell counts by 22.67% and 44.92%, respectively. In addition, compared with CSA treatment, treatment ADR 200 mg/kg reduced neutrophils, CD4+/CD3+ cells and CD8+/CD3+ cells, by 32.10%, 83.17% and 82.11%, respectively. These results indicate that ADR extract may have an inhibitory effect on COPD induced by LPS and CSC in mice.

This study was carried out to investigate whether Chrysanthermum zawadskii var. latilobum Kitamura (C. zawadskii) extracts has an inhibitory effect against the mutagenicity by cigarette smoke condensates (CSC). C. zawadskii was extracted with 70% ethanol and the yield was 18.5%. We further fractioned 70% ethanol extract sequentially to diethylether, chloroform, dichloromethane, and aqueous water, and gained the yield of 17.5%, 5.6%, 5.8%, 32.8% and 35.5%, respectively. In the Ames test, there was no mutagenic effect of crude extract and its solvent fractions up to 2 mg/plate toward Salmonella typhimurium TA 98 with or without S-9 mix metabolic activations. On the contrary, the crude extract showed an inhibitory activity against the mutagenicity of CSC in the presence of S-9 mix metabolic activation. Diethyl ether layer among five solvent fractions showed the highest inhibitory activity. The inhibitory activity of diethyl ether fraction was also increased in a dose-dependent manner and the inhibitory rate was about 97.7% at the concentration of 1 mg/plate. In this study, we conclude that crude extract of C. zawadskii itself is potentially safe for mutagenicity, and the diethyl ether fraction has an inhibitory effect against the mutagenicity of CSC.

This study was carried out to investigate the antimicrobial activities of organic extracts obtained from the fruit of Thuja orientalis L. The native fruits in Korea were collected and extracted by 80% ethanol, and the extract was sequentially fractionated with n-hexane, chloroform, ethylacetate, and butanol. The fraction yields of n-hexane, chloroform, ethylacetate, butanol and water of ethanol extract were 10.15%, 10.05%, 1.45%, 45.35% and 27.55%, respectively. n-Hexane-soluble fraction showed the highest antibacterial activity against gram positive bacteria, while the chloroform, ethylacetate, butanol and aqueous fractions did not show any antibacterial activity. Minimum inhibitory concentration (MICs) on Staphyloycoccus aureus, Bacillus subtilis and Sateptococcus pneumoniae, n-hexane-soluble fraction were 100 μg, 500 μg and 50 μg/disc, respectively. The antibacterial activity was not destroyed by heating at 80, 100, 120℃ for 30 min and was not affected by pH. In the inhibitory test against the Staphylococcus aureus, Bacillus subtilis and Sateptococcus pneumoniae, n-hexane-soluble fraction showed potent growth inhibition at the concentration of 0.1 and 0.5 μg/mL for 12~24 hours and n-hexane-soluble fraction did not show any mutagenic activity.

The objective of this study was to investigate the effect of crude ginseng total saponins (CGS) against airway inflammation (AI) and airway hyperresponsiveness (AH) induced by diesel exhaust particles (DEP) in mice. AI and AH were induced by the intratracheal instillation with 0.1 mg/ml of DEP suspension once a week for 10 weeks combined with ovalbumin (OVA) sensitization. Mice were also treated orally with 75 mg/ml of CGS, 5 days a week for 10 weeks. Oral CGS treatment decreased in the level of serum immunoglobulin (IgE) and histamine increased by DEP and OVA, and declined respiratory resistance. It also dropped an enhanced infiltration of eosinophils in the bronchoalveolar lavage fluid (BALF) of mice, and an increased T helper type 2 cell derived cytokine levels such as of interleukin (IL)-4, IL-13 and IL-5 in the BALF. However, it did not influence T helper type 1 cytokine such as interferon-gamma in the BALF. These results indicate that CGS may alleviate allergen-related AI and AH in mice and may play an important role in the modulation of asthmatic inflammation.

홍삼의 물추출물을 장기간 흰쥐에 급여했을 때 흰쥐 간의 이물질대사효소 활성도 유도에 미치는 영향을 조사하였다. 홍삼 물추출물의 급여농도는 25 mg/kg body weight이었으며, 간의 이물질대사효소 유도제는 항경련제 PB (80 mg/kg)와 발암제 3-MC (20 mg/kg)을 사용하였다. 간에서 이물질대사에 관여하는 cytochrome P-450의 함량, cytochrome P-450 dependent monooxygenase인 ECOD와 BPDM, cytochrome P-450 reductase와 GST의 활성도는 PB와 3-MC 투여에 의해서 대조군과 홍삼투여군 모두 크게 증가되었다. 흰쥐의 나이가 3개월에서는 두 유도제를 투여했을 때 이물질대사효소의 활성은 대조군이 인삼투여군에 비해 상대적으로 더 많이 유도되었다. 그러나 12개월에서는 두 시험군 모두 유도정도가 감소하였으나 홍삼투여군은 그 활성도 감소정도가 대조군에 비해 상대적으로 크지 않았다. 한편 두 그룹 모두 이물질대사 효소 중 cytochrome B5 함량과 cytochrome b5 reductase의 활성도는 PB와 3-MC 투여에 의해서 뚜렷한 차이를 보이지 않았다. 따라서 이러한 결과는 흰쥐에 홍삼의 장기간 급여는 간에서 효소유도제의 이물질대사효소유도 정도를 조절하여 무독화에 기여할 수 있다는 것을 의미한다.

The present study was designed to investigate aging-related effects on the activities of xenobiotic metabolizing enzymes of rat liver by dietary supplementation of Korean red ginseng water-extract. Rat did not show any discernible signs of the rejection symptoms, and blood GOT and GPT levels were not influenced by ginseng water extracts, Cytochrome 450 levels and NADPH cytochrome P45O reductase, p-450 dependent ethoxycoumarin O-deethylase, and benzphetamine N-demethylase activities were decreased with aging, however, these phase I system enzymes activities in the ginseng group of24 months were well maintained compared with normal group. But, Levels of cytochrome bs and NADH-cytochrome b5 reductase activities were also decreased with aging and were not found a clear difference between two groups. Glutathione-s-transferase activity, phase II enzyme system, in liver cytosols was also decreased in old ages, but the degree of decrease was higher in normal group than in giuseng supplemented group. These results indicate that long-term supplementation of red ginseng water extracts from weaning to 24 months do not show any side effects to rats, and retard age-related deteriorations of xenobiotic metabolizing enzymes activities in old ages.