Folk Medicine In Taiwan Research Articles

Phellinus linteus mycelia extract in COVID-19 prevention and identification of its key metabolic compounds profiling using UPLC-QTOF-MS/MS spectrometry

For centuries, food, herbal medicines, and natural products have been valuable resources for discovering novel antiviral drugs, uncovering new structure-activity relationships, and developing effective strategies to prevent/treat viral infections. One such resource is Phellinus linteus, a mushroom used in folk medicine in Taiwan, Japan, Korea, and China. In this rich historical context, the key metabolites of Phellinus linteus mycelia ethanolic extract (GKPL) impacting the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at multiple stages have yet to be explored. Thus, this study systematically identifies and assesses the inhibitory effect of GKPL on the SARS-CoV-2 virus. Initially, the concentrations and contact times of GKPL against SARS-CoV-2 pseudovirus were assessed in HepG2 cells. Subsequently, utilizing the Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry method, potential biomarkers in the fungal extract were discerned. Metabolomic analysis identified 18 compounds in GKPL, with hispidin and hypholomine B present in the highest amounts. These compounds were isolated using chromatographic techniques and further identified through 1D NMR spectroscopic and mass spectrometry analysis. Hispidin and hypholomine B were found to inhibit the infection of SARS-CoV-2 pseudovirus by reducing angiotensin-converting enzyme 2 gene expression in HepG2, thereby decreasing viral entry. Moreover, hispidin and hypholomine B effectively block the spike receptor-binding domain, while hypholomine B, for the first time, showed significant inhibition of 3CL protease. This suggests that GKPL, enriched with hispidin and hypholomine B, has the potential to be used as an active ingredient against SARS-CoV-2.

Analysis of flavonoids in Vernonia paltula by high-performance liquid chromotography

Lin-nan-yeh-chu, is the dried entire plant of Vernonia paltula (Compositae) and used as folk medicine in Taiwan. To evaluate the quality of V. paltula, a simple, rapid and accurate high-performance liquid chromatographic (HPLC) method was developed for the assay of four flavonoids apigenin (API), apigenin-7-O-glucoside (APG), luteolin (LUT) and luteolin-7-O-glucoside (LUG). The present HPLC system uses an Inertsil ODS-2 column by gradient elution with acetonitrile and 0.1% (v/v) phosphoric acid as the mobile phase. Ethyl paraben was used as an internal standard and detected at 254 nm. Regression equations revealed good linear relationships (correlation coefficients: 0.9998-0.9999) between the peak-area ratios of each constituent to ethyl paraben. The recovery of four marker constituents ranged from 89.3 to 95.6%. The contents of the four constituents in stem, flower, leaf and root parts of V. paltula have been compared. Leaf part consisted of the highest contents of flavonoids except for APG which is less than that in the flower. The root and stem only showed trace amount of APG but not the other three flavonoids. The contents of four constituents were 0.6016, 0.0042, 0.2160 and 0.0577 mg/g for apigenin-7-O-glucoside, apigenin, luteolin-7-O-glucoside and luteolin, respectively. HPLC chromatograms of another three plants of genus of Vernonia in Taiwan, V. cinerea, V. elliptica and V. gratiosa, have also been compared.

Cyclohexylamine, an active compound from Toddalia asiatica, contracts epididymal vas deferens via serotonergic receptors.

BackgroundToddalia asiatica of Rutaceae, a Taiwan folk medicine, is used as an analgesic and anti-inflammatory herb. Cyclohexylamine (CHA) is an active compound from T. asiatica. Previous reports indicate CHA contracts rat vas deferens. However, the contractile mechanism of CHA on rat vas deferens was not yet reported. The purpose of this study was to investigate the contractile mechanism of CHA on rat epididymal portion of vas deferens.MethodsMale S.D. rats weighting between 200 g to 250 g were used. The epididymal portion of vas deferens was isolated and was added with various concentrations of serotonin, serotonin antagonists and CHA.ResultsSerotonin elicited dose-dependent (1 × 10−7M~1 × 10−4M) contractions on rat epididymal vas deferens, which were inhibited by pretreatment with ketanserin (1 × 10−8 M ~ 1 × 10−6 M), methysergide (1 × 10−5 M) and propranolol (1 × 10−4 M), respectively. CHA elicited dose-dependent (1 × 10−8M~1 × 10−4M) contractions on rat epididymal vas deferens. The contractions of CHA (1 × 10−4M) on epididymal vas deferens were enhanced by serotonin in a dose-dependent manner. Methysergide (1 × 10−7 ~1 × 10−5 M) did not affect the contractions evoked by CHA. However, the CHA elicited contraction was almost completely inhibited by ketanserin (1 × 10−5 M) and was enhanced by propranolol. The effect of propranolol at the concentration of 1 × 10−4 M on CHA was likely as CHA at high concentration alone.ConclusionsFrom the above results, the contraction evoked by CHA on the isolated rat epididymal vas deferens might be mediated by serotonergic receptors through 5-HT2A subtype.

MicroRNA-like small RNAs prediction in the development of Antrodia cinnamomea.

Antrodia cinnamomea, a precious, host-specific brown-rot fungus that has been used as a folk medicine in Taiwan for centuries is known to have diverse bioactive compounds with potent pharmaceutical activity. In this study, different fermentation states of A. cinnamomea (wild-type fruiting bodies and liquid cultured mycelium) were sequenced using the next-generation sequencing (NGS) technique. A 45.58 Mb genome encoding 6,522 predicted genes was obtained. High quality reads were assembled into a total of 13,109 unigenes. Using a previously constructed pipeline to search for microRNAs (miRNAs), we then identified 4 predicted conserved miRNA and 63 novel predicted miRNA-like small RNA (milRNA) candidates. Target prediction revealed several interesting proteins involved in tri-terpenoid synthesis, mating type recognition, chemical or physical sensory protein and transporters predicted to be regulated by the miRNAs and milRNAs.

Inhibitory effects of wild bitter melon leaf extract on Propionibacterium acnes-induced skin inflammation in mice and cytokine production in vitro.

Propionibacterium acnes is a key pathogen involved in acne inflammation. Wild bitter melon (WBM, Momordica charantia L. var. abbreviate Seringe) is consumed as both a vegetable and as folk medicine in Taiwan. We examined the inhibitory activity of the total phenolic extract (TPE) of WBM leaf on P. acnes-induced inflammatory responses in vivo and in vitro. Our data showed that TPE significantly attenuated P. acnes-induced ear swelling in mice along with microabscess. Flow cytometry analysis revealed that TPE treatment significantly decreased the migration of neutrophils and interleukin (IL)-1β(+) populations in vivo. In P. acnes-stimulated human monocytic THP-1 cells, TPE suppressed the mRNA levels and production of IL-8, IL-1β, and tumor necrosis factor (TNF)-αin vitro. In addition, TPE suppressed P. acnes-induced matrix metalloproteinase-9 levels. TPE blocked nuclear factor-κB (NF-κB) activation and inactivated mitogen-activated protein kinases (MAPK); these actions may partially account for its inhibitory effect on cytokine production. The quantitative HPLC analysis revealed gallic, chlorogenic, caffeic, ferulic, and cinnamic acids, myricetin, quercetin, luteolin, apigenin, and thymol in TPE. All these phenolics significantly suppressed P. acnes-induced IL-8 production in vitro. Our results suggest that WBM leaf extract effectively inhibits P. acnes-induced inflammatory responses and may be useful to relieve the inflammation of acne.

Bioactive constituents of Cirsium japonicum var. australe.

Cirsium japonicum var. australe, used as a folk medicine in Taiwan, has been employed traditionally in the treatment of diabetes and inflammatory symptoms. Bioactivity-guided fractionation of its ethanolic extract, utilizing centrifugal partition chromatography monitored by DPPH-TLC analysis, led to the isolation of three new acetylenic phenylacrylic acid esters (1-3) and two new polyacetylenes (4 and 5), together with seven known compounds (6-12). The structures of 1-5 were elucidated by spectroscopic methods including 1D and 2D NMR techniques. The absolute configurations of 4 and 7 were determined utilizing Mosher's method and ECD/CD experiments. The DPPH scavenging activity of the constituents isolated from the C. japonicum var. australe ethanolic extract was evaluated. The potential antidiabetic activity of some of the isolates was evaluated using in vitro cellular glucose uptake and oil red staining assays.

Vitis thunbergii supplementation demonstrates an anti-obesity effect in developing obese mice

IntroductionVitis thunbergii (VTE), an indigenous wild grape of Taiwan, has been widely used as folk medicine in Taiwan. In this study, we investigated the anti-obesity effect and safety of VTE in high-fat diet-fed C57BL/6JNarl mice. Materials and methodsMale C57BL/6JNarl mice were randomly divided into three groups fed with diets for 7 weeks: a high-fat diet (control group), high-fat diet with 0.375% V. thunbergii extract (1× VTE group), and high-fat diet with 0.75% V. thunbergii extract (2× VTE group). Body weight and food intake were recorded. The resected epididymal fat, liver, and kidney were collected for further analysis. ResultsOur data suggested an anti-obesity effect of VTE in high-fat diet-fed C57BL/6JNarl mice, as evidenced by the decreased body weight gain, epididymal fat pads weight, adipocyte size, liver weight, and hepatic fat deposition. Moreover, VTE supplementation lowered the level of blood glucose and serum triglyceride. However, histological changes and parallel elevation of serum GOT and GPT level demonstrated a potential hepatotoxicity in 0.75% VTE-supplemented group. The augmented liver injury was accompanied by fibrotic change, possibly through activation of α-SMA-positive matrix-producing cells. ConclusionsThe findings suggest that VTE supplementation showed anti-obesity effect and preferentially ameliorated glucose and lipid dysregulation in C57BL/6JNarl mice fed with a high-fat diet. However, the long-term use of VTE should be cautioned because of a potential hepatotoxicity, especially at higher doses.

Supplementation of Vitis thunbergii root extract alleviated high-fat diet-induced obesity in C57BL/6J mice

Vitis thunbergii root, widely used as folk medicine in Taiwan, has been found to contain polyphenolic compounds and resveratrol derivatives, which have been implicated in the prevention and treatment of obesity. Thus, we hypothesized it might show beneficial effects against obesity. C57BL/6JNarl mice fed with a high fat diet for 14 weeks increased body weight and epididymal fat pad weight, and accompanied by fatty liver, hyperglycemia, hyperinsulinemia, insulin resistance, hyperleptinemia, hypercholesterolemia, hyper-LDL-cholesterol, and high level of serum GPT, GOT, creatinine, and BUN. Supplementation of VTE in the last 7 weeks remarkably decreased body weight and epididymal fat pad weight, implying a potential anti-obesity effect. Mechanistic study showed that VTE supplementation increased energy expenditure-related CPT1 mRNA expression and AMPK phosphorylation, and decreased lipogenesis-related SREBP-1 expression in liver. In conclusion, Vitis thunbergii roots could alleviate high fat diet-induced obesity and its related complications by enhancing hepatic fatty acid oxidation and inhibitng lipogenesis.

Cytotoxicity of (-)-vitisin B in human leukemia cells

Vitis thunbergii var. taiwaniana (VTT) is an indigenous Taiwanese wild grape and is used as a folk medicine in Taiwan. VTT is rich in polyphenols, especially quercetin and resveratrol derivatives, which were demonstrated to exhibit inhibitory activities against carcinogenesis and prevent some neurodegenerative diseases. (-)-Vitisin B is one of the resveratrol tetramers extracted from VTT. In this study, we investigated the mechanisms of (-)-vitisin B on the induction of apoptosis in human HL-60 promyelocytic leukemia cells. First, (-)-vitisin B significantly inhibited cell proliferation through inducing cell apoptosis. This effect appeared to occur in a time- and dose-dependent manner. Cell-cycle distribution was also examined, and we found that (-)-vitisin B significantly induced a sub-G1 population in a dose-dependent manner. In addition, (-)-vitisin B exhibited stronger inhibitory effects on cell proliferation than resveratrol. Second, (-)-vitisin B dose dependently induced apoptosis-related protein expressions, such as the cleavage form of caspase-3, caspase-8, caspase-9, poly(ADP ribose) polymerase, and the proapoptotic Bax protein. Third, (-)-vitisin B treatment also resulted in increases in c-Jun N-terminal kinase (JNK) phosphorylation and Fas ligand (FasL) expression. Moreover, the (-)-vitisin B-induced FasL expression and caspase-3 activation could be reversed by a JNK inhibitor. These results suggest that (-)-vitisin B-induced apoptosis of leukemia cells might be mediated through activation of JNK and Fas death-signal transduction.

Anti-Inflammatory molecular mechanisms of Bo-Ye-Niu-Pi-Xiao (Cynanchum taiwanianum Yamazaki)

Bo-Ye-Niu-Pi-Xiao, Cynanchum taiwanianum Yamazaki (family: Asclepiadaceae) is a well-known and popular herb; its rhizome has been used as a folk medicine in Taiwan. Many therapeutic effects of C. taiwanianum Yamaz. had been studied; however, there is still no anti-inflammatory effect and mechanism of C. taiwanianum Yamaz. reported. Besides, it is well-known that the phosphorylation of I kappa B-alpha (IκBα) to phospho-IκBα (p-IκBα) is a decisive step in the nuclear factor kappa B (NF-κB) activation pathway. In addition, the induction of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) by cytokines, such as interleukin-1β (IL-1β), is well known to be partly mediated by NF- κB. Hence, the objective of this study was to evaluate the activity and distinguish the mechanism of anti-inflammation of C. taiwanianum Yamaz. rhizome on rat kidney cell line NRK-52E induced by IL-1β. In the present study, we investigated the anti-inflammatory effect and potential mechanism of C. taiwanianum Yamaz. rhizome ethanolic extract (CTEE) in IL-1β-induced NRK-52E cells. Production of nitric oxide (NO) and prostaglandin E2 (PGE 2 ) by enzyme linked immunosorbent assay. The messenger ribonucleic acid (mRNA) and protein expression of iNOS and COX-2, phosphorylation of IκBα, and activation of NF-κB was executed by reverse transcription-polymerase chain reaction and Western blotting. Results show that the CTEE significantly (P < 0.05) inhibited NO and PGE 2 production and also significantly (P < 0.05) attenuated protein and mRNA expression of iNOS and COX-2 in IL-1β-induced NRK-52E cells without obvious cytotoxic effects. Furthermore, the CTEE suppressed the NF-κB nuclear translocation, in terms of inhibition of IκBα phosphorylation. The results provided evidence for its folkloric uses and suggest that the anti-inflammatory activities of CTEE may result from the inhibition of inflammatory mediators, such as NO and PGE 2 and an upstream suppression of NF-κB-dependent mechanism might be involved. Key words : Cynanchum taiwanianum Yamazaki, anti-inflammatory, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS).

First total synthesis of antrocamphin A and its analogs as anti-inflammatory and anti-platelet aggregation agents

Naturally occurring antrocamphin A (1) is a potent anti-inflammatory compound from the edible fungus Antrodia camphorata (Taiwanofungus camphoratus), whose wild fruiting body is used as a valuable folk medicine in Taiwan. This study is the first total synthesis of antrocamphin A (1) and its analogs. Their inhibition ability on NO release, superoxide anion generation, elastase release and platelet aggregation are reported herein.

Evaluation of antioxidant and DNA protection activities in the extracts of the Terminalia catappa leaves

Terminalia catappa (Combretaceae) is used commonly as a folk medicine in Taiwan, and has been claimed to have therapeutic effects in hepatitis and liver related diseases [1, 2]. The leaves, bark and fruit of the Terminalia catappa have also been commonly used as a folk medicine for antidiarrheic, antipyretic and haemostatic purposes in India, Philippines, Malaysia and Indonesia [3]. The extracts from the Terminalia catappa leaves were used in this study. Antioxidant activities were measured by both 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH·) scavenging and 2, 2-azino-bis (3-ethylbenz- thiazoline-6-sulfonic acid) (ABTS·+) decolourisation methods. To further evaluate the effect of Terminalia catappa extracts on UV induced DNA damages, the DNA protection assay was employed. Terminalia catappa leaf extracts had effective DPPH bleaching activity and ABTS·+ radical scavenging activity in a concentration dependent manner. Furthermore, the Terminalia catappa leaf extracts to inhibit the oxidative DNA damages were assessed by measuring the conversion of supercoiled pUC119 plasmid DNA to the linear forms. UV irradiation of DNA with hydrogen peroxide resulted in the formation of linear forms of DNA, indicating double-strand DNA breaks. Addition of Terminalia catappa plant extracts to DNA resulted in a partial inhibition of the conversion of supercoiled DNA to linear forms, indicating that the Terminalia catappa plant extract was able to protect plasmid DNA against hydroxyl radical induced oxidative damage. The inhibition of hydroxyl radical induced DNA strand breaks by Terminalia catappa plant extracts exhibited a concentration dependent relationship.

Abstract 780: Dimethyl cardamonin induces G1-phase cell cycle arrest, apoptosis, and autophagy in HCT116 cells

Abstract Dimethyl cardamonin (2’,4’-dihydroxy-6’-methoxy-3’,5’-dimethylchalcone; DMC) is a naturally occurring chalcone, and it is the major compound isolated from the leaves and/or fruits of Syzygium samarangense (Blume) Merr. &amp; L.M. Perry (Myrtaceae). DMC has been reported to have anti-diabetic, anti-bacterial, spasmolitic and anti-tumor properties. Furthermore, it has been used as an antipyretic and a diuretic agent in Taiwan folk medicine. However, the underlying mechanisms of its anti-tumor effects have not been studied. Here, we report that DMC induced G1 phase arrest, apoptosis and autophagy in HCT116 cell line. In this study, DMC decreased the expression of the phospho-Rb, CDK4 and cyclin D1 expression level, which is correlated with G1 phase. We also found that DMC-induced apoptosis occurred through the p53 dependent manner and the mitochondrial death pathway as evidenced by reduction of the Bcl2, XIAP and caspase-9 activation, and by induction of Bims. And, we showed that DMC up-regulates activity of JNK and p38. Interestingly, we found that DMC elicits autophagy, a process that participates in apoptotic response, in a JNK and p53-dependent manner, in HCT116 cell line. We also found that the increased activation of JNK by DMC is partially regulated by p53, highlighting the relationship of JNK and autophagy to p53 signaling pathway. The treatment of HCT116 (p53 -/-) cell line with DMC resulted in reduction in both apoptotic and autophagic effects relative to HCT116 (p53 +/+) cell line. This study suggested that DMC induced the multiple effects of G1 phase arrest, apoptosis and autophagy through JNK and p53 in HCT 116 cell line. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 780.

Osteoclast-Forming Suppressive Compounds from Anoectochilus formosanus

Anoectochilus formosanus, an orchidaceous plant native to Taiwan, is widely used as a folk medicine in Taiwan and other Southeast Asian countries. We attempted to isolate the bioactive principles from A. formosanus and succeeded in purifying the compounds. Two known compounds (N-cis-feruloyl tyramine and N-trans-feruloyl tyramine) were isolated and both significantly inhibited osteoclast formation, which is important in the development of osteoporosis.

Extract isolated from Angelica hirsutiflora with insulin secretagogue activity

Ethnopharmacological relevance Angelica genus (Umbelliferae) has traditionally been used as the medicine and health food considered alleviating several disorders including diabetes mellitus. Angelica hirsutiflora Liu Chao & Chuang is an endemic species and a folk medicine in Taiwan. Aim of the study The scientific evidence of anti-diabetic effect for Angelica hirsutiflora remains unknown. The methanolic extracts isolated from Angelica hirsutiflora were studied for its insulin secretagogue and hypoglycemic activities. Materials and methods The in vitro effects and possible mechanisms of Angelica hirsutiflora extract on the insulin secretion in isolated mouse and human islets and pancreatic β-cell line HIT-T15 were determined; and tested the regulation of blood glucose in fasted mice and high-fat diet-induced diabetic mice. Results Angelica hirsutiflora extract potently stimulated the release of insulin from cultured HIT-T15 cells and isolated mouse and human islets. The intracellular calcium levels were also increased in HIT-T15 cells and isolated human islets treated with Angelica hirsutiflora extract. Angelica hirsutiflora extract was capable of enhancing the phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2 protein in HIT-T15 cells. Specific ERK inhibitor PD98059 inhibited the increase of insulin secretion by Angelica hirsutiflora extract in HIT-T15 cells and isolated mouse islets. When Angelica hirsutiflora extract was administered to the fasted mice, it decreased the rise in blood glucose level after starch loading. The plasma insulin level was also increased by Angelica hirsutiflora extract treatment. In high-fat diet-induced diabetic mice, Angelica hirsutiflora extract markedly improved the oral glucose intolerance as compared with the vehicle control. Conclusions These findings support that Angelica hirsutiflora extract may be useful in the control of hyperglycemia in non-insulin-dependent diabetes mellitus by acting as an insulin secretagogue.

The Augmented Anti-Tumor Effects of Antrodia camphorata Co-Fermented with Chinese Medicinal Herb in Human Hepatoma Cells

Antrodia camphorata, unique fungal specie, has been used as a folk medicine in Taiwan for many years. The purpose of this study was to compare the extracts from the solid-state culture of A. camphorata co-fermented with Chinese medicinal herb (AC-CF) with two other extracts from fruiting bodies (AC-FB) or solid-state culture (AC-SS), for their anti-tumor effects in human hepatoma HepG2 cells. We measured in vitro cell proliferation, percentage of apoptosis, population distribution of cell cycles, Western blot analysis of multiple drugs resistance-1 (MDR-1), and apoptosis-related proteins in HepG2 cells treated with three different preparations of A. camphorate extracts. Our results showed that AC-CF had better anti-proliferation effect on human hepatoma HepG2 cells than AC-FB or AC-SS dose-dependently. In addition, AC-CF in combination with anti-tumor agents (mitomycin C or methotrexate) showed better adjuvant anti-tumor effects than AC-FB or AC-SS. We further demonstrated the augmented adjuvant anti-tumor effects of AC-CF not only through down regulation of MDR-1 expression but also through a COX-2 dependent apoptosis pathway, involving down-regulation of COX-2 and p-AKT and up-regulation of PARP-1. In conclusion, in this study, we have demonstrated a novel strategy of fermenting A. camphorata with Chinese medicinal herb (AC-CF), which augmented their anti-tumor effects in human hepatoma HepG2 cells as compared to the traditional ones (AC-FB or AC-SS).

Demographics, Training, and Practice Patterns of Practitioners of Folk Medicine in Taiwan: A Survey of the Taipei Metropolitan Area

The purpose of this study was to investigate the demographics, training, and practice patterns of folk medicine practitioners, their opinions toward statutory regulation of folk medicine, and the formal education and credentialing for folk medicine providers in the metropolitan Taipei area. A cross-sectional survey design was used. Included in the survey were 200 folk medicine practitioners in Taipei city and 200 folk medicine practitioners in Taipei county. The survey questionnaire consisted of 3 domains including demographics and training; practice patterns; and opinions toward statutory regulation of folk medicine and formal education and credentialing for Tuina, Ba Guan, Gua Sha, and reflexology providers. The response rates ranged from 86.3% to 99.5%. A typical folk medicine provider in the Taipei metropolitan area was a middle-aged man with a high school degree who worked about 50 hours a week. The majority of the providers in the Taipei metropolitan area received their training through apprenticeship. Years of training and experience varied widely among these practitioners. About 80% had received more than one year of training prior to starting their practice. Adult men and women were their major clientele. The major treatment modalities they offered were Tuina, Gua Sha, Ba Guan, reflexology, and meridian massage. The majority of the respondents agreed that practitioners should receive formal education and training and agreed that certifying the qualifications of folk medicine practitioners is necessary. Findings from the present survey provide an understanding of the training and practice patterns of Taiwanese folk medicine practitioners, highlight folk medicine practitioners' needs for formal education and training, and stress the importance of statutory regulation of folk medicine in Taiwan.

Antibacterial activity of Melastoma candidum D. Don

Melastoma candidum D. Don is a plant of the Melastomataceae family that has been used as folk medicine in Taiwan. In this study, the antibacterial activity of M. candidum extracts was investigated. Water, acetone, ethanol (95 ml/100 ml), and ethyl acetate extracts of M. candidum exhibited antibacterial activity, especially the acetone and ethanol (95 ml/100 ml) extracts. Over one-third of MICs for those two extracts were close to or equal to that of amoxicillin. MICs and minimum bactericidal concentrations (MBCs) for the acetone extract were 0.02–0.64 mg/ml and 0.08–2.56 mg/ml, respectively; while for the ethanol (95 ml/100 ml) extract, corresponding values were 0.04–1.28 mg/ml and 0.16–5.12 mg/ml, respectively. The acetone M. candidum extract exhibited good thermal stability subsequent to heating from 70 to 121 °C for 15–60 min; further, no dramatic changes in the MICs in a range of pH 5–8 were noted. Overall, M. candidum acetone extract revealed a good bactericidal effect, good thermal stability (heating at 121 °C for 15 min), and broad antibacterial activity in the pH range of 5–8.

生薬学研究40年

In order to quality control for medicinal plants such as Aconitum charmicaelii, Rhemannia glutinosa, Atractyrodes lancea, Pinellia ternata, Panax species and Gentiana scabra were clonally micropropagated by tip tissue culture and embryogenetic techniques. Monoclonal antibodies against the bioactive compounds contained in the crude drugs were prepared and set up the ELISA as a high sensitive and quick determination method. A newly developed eastern blotting methodology can stain typically the only antigen molecule and the related compounds having a same aglycone. Knockout extract can be prepared by using an immunoaffinity column conjugated with monoclonal antibody, and its importance has been discussed. Single chain fragment-variable gene against solamargine was cloned and transformed to a host plant, Solanum chasianum resulting in the increase of antigen molecule as 2.5 to 3 times. Three biosynthetic enzymes regarding marihuana compounds, THCA-, CBDA-, CBCA-synthases were isolated. THCAsynthase was cloned, over expressed and confirmed its characteristics including FAD combining enzyme, and finally determined the structure by X-ray analysis. The distribution of THCAsynthase was also investigated using its and GFP hybrid gene. We found new functions for saffron like improving learning and memory and LTP for blocking by ethanol. A folk medicine in Taiwan, Anoectochilus formosanus was propagated in vitro and investigated opening new pharmacological activities in lipid metabolism.

Mechanisms of suppression of nitric oxide production by 3- O-methylquercetin in RAW 264.7 cells

Rhamnus nakaharai Hayata (Rhamnaceae) is used as a folk medicine in Taiwan for treating constipation, inflammation, tumors, and asthma. 3- O-Methylquercetin (3-MQ), a main constituent of the plant, has been reported to have potential for use in the treatment of asthma. The mechanisms of anti-inflammation of 3-MQ are still unclear. Nitric oxide (NO) production induced by lipopolysaccharide (LPS) through iNOS expression in RAW 264.7 cells, a mouse macrophage cell line, may reflect the degree of inflammation and may provide a measure for assessing the effect of drugs on the inflammatory process. Therefore, we were interested in investigating the mechanisms of suppression of NO production by 3-MQ in RAW 264.7 cells. 3-MQ (1–10 μM) concentration-dependently inhibited LPS (100 ng/mL)-induced NO production in RAW 264.7 cells. The IC 50 value was calculated to be 4.23 μM. 3-MQ (1–10 μM) significantly and concentration-dependently inhibited LPS (100 ng/mL)-induced iNOS protein and mRNA expressions in cells. The IC 50 values were calculated to be 4.36 and 6.53 μM, respectively. There was no significant difference among these three IC 50 values of 3-MQ. In conclusion, 3-MQ may exert its anti-inflammatory effect through the inhibition of iNOS DNA transcription.