Weena Jiratchariyakul

Inhibition of Wnt signaling by cucurbitacin B in breast cancer cells: Reduction of Wnt associated proteins and reduced translocation of galectin-3-mediated β-catenin to the nucleus

The cucurbitacins are tetracyclic triterpenes found in plants of the family Cucurbitaceae. Cucurbitacins have been shown to have anti‐cancer and anti‐inflamatory activities. We investigated the anti‐cancer activity of cucurbitacin B extracted from Thai medicinal plant Trichosanthes cucumerina Linn. Cell viability was assessed by MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide) assay. Results indicated that cucurbitacin B from T. cucumerina Linn. has a cytotoxic effect on breast cancer cell lines SKBR‐3 and MCF‐7 with an IC50 of 4.60 and 88.75 µg/ml, respectively. Growth inhibition was attributed to G2/M phase arrest and apoptosis. Cyclin D1, c‐Myc, and β‐catenin expression levels were reduced. Western blot analysis showed increased PARP cleavage and decreased Wnt‐associated signaling molecules β‐catenin, galectin‐3, cyclin D1 and c‐Myc, and corresponding changes in phosphorylated GSK‐3β levels. Cucurbitacin B treatment inhibited translocation to the nucleus of β‐catenin and galectin‐3. The depletion of β‐catenin and galectin‐3 in the nucleus was confirmed by cellular protein fractionation. T‐cell factor (TCF)/lymphoid enhancer factor (LEF)‐dependent transcriptional activity was disrupted in cucurbitacin B treated cells as tested by a TCF reporter assay. The relative luciferase activity was reduced when we treated cells with cucurbitacin B compound for 24 h. Our data suggest that cucurbitacin B may in part induce apoptosis and exert growth inhibitory effect via interruption the Wnt signaling. J. Cell. Biochem. 113: 49–60, 2012.

Cytotoxic properties of root extract and fruit juice of Trichosanthes cucumerina.

The root extract of Trichosanthes cucumerina L. and bryonolic acid (1), its main constituent, as well as the fruit juice and cucurbitacin B (3), its main constituent, were tested for cytotoxicity against four human breast cancer cell lines (SKBR3, MCF7, T47D, and MDA-MB435), two lung cancer cell lines (A549 and SK-LU1), and one colon cancer cell line (Caco-2). The root extract had higher IC (50) values than bryonolic acid (1) against three breast cancer cell lines (MCF7 = 267/121, T47D = 316/124, MDA-MB435 = 140/90 microL/mL) and one lung cancer cell line (A549 = 106/100 microL/mL). The fruit juice also had higher IC (50) values than cucurbitacin B (3) against the four breast cancer cell lines (131/73, 375/35, 249/60, and 156/26 microL/mL, respectively) and one lung cancer cell line (141/41 microL/mL) as shown above, as well as against the colon cancer cell line (101/1.5 microL/mL). However, the root extract inhibited SK-LU1 more strongly than did the fruit juice, cucurbitacin B (3), and bryonolic acid (1) (149/169/180/>500 microL/mL, respectively). The root extract inhibited the two lung and three breast cancer cell lines (SKBR3, MDA-MB435, and MCF7) more strongly than the fruit juice. Bryonolic acid (1) inhibited MDA-MB435 somewhat better than the other tested human cancer cell lines. The fruit juice inhibited the colon cancer cell line (Caco-2) more strongly than the root extract. Cucurbitacin (3) inhibited human cancer cell lines, especially Caco-2, much more strongly than bryonolic acid (1). In addition to bryonolic acid (1), bryononic acid (2), cucurbitacin B (3), and dihydrocucurbitacin B (4) also were isolated from the root extract.

Antiproliferative Effects of Cucurbitacin B in Breast Cancer Cells: Down-Regulation of the c-Myc/hTERT/Telomerase Pathway and Obstruction of the Cell Cycle

Naturally occurring cucurbitacins have been shown to have anticancer, antimicrobial and anti-inflammatory activities. In this study, we determined the effects of cucurbitacin B extracted from the Thai herb Trichosanthes cucumerina L. on telomerase regulation in three human breast cancer cell lines (T47D, SKBR-3, and MCF-7) and a mammary epithelium cell line (HBL-100). Cell viability after treatment with cucurbitacin B, which is an active ingredient of this herb, was assessed. Telomeric Repeat Amplification Protocol (TRAP) assays and RT-PCR (qualitative and realtime) were performed to investigate activity of telomerase as well as expression of human telomerase reverse transcriptase (hTERT) and c-Myc. The c-Myc protein level was also determined in SKBR-3 and HBL-100 cells. Our results show that the cucurbitacin B inhibits growth and telomerase activity in the three breast cancer cell lines and exerts an obvious inhibitory effect in the estrogen receptor (ER)-negative breast cancer SKBR-3 cells. The expression of hTERT and c-Myc were also inhibited by cucurbitacin B, In addition, a clear reduction of c-Myc protein was observed after treatment in SKBR-3 cells even with a concentration of cucurbitacin B that was ten-times lower compared to the concentration used for HBL-100. Our findings imply that cucurbitacin B exerts an anticancer effect by inhibiting telomerase via down regulating both the hTERT and c-Myc expression in breast cancer cells.

Overview of Botanical Status in EU, USA, and Thailand

The botanical status in EU, USA, and Thailand is different owing to the regulatory status, the progress of science, and the influence of culture and society. In the EU, botanicals are positioned as herbal medicinal products and food supplements, in the US they are regulated as dietary supplements but often used as traditional medicines, and in Thailand, they are regulated and used as traditional medicines. Information for some of the most popular botanicals from each country is included in this review.

Antibacterial and Antiproliferative Activities of Plumericin, an Iridoid Isolated from Momordica charantia Vine

Plumericin, an iridoid lactone, was isolated with relatively high yield from Momordica charantia vine using the supercritical fluid extraction (SFE) and the separation box (Sepbox) comprising dual combination of high-performance liquid chromatography and solid phase extraction. This compound showed antibacterial activity against Enterococcus faecalis and Bacillus subtilis with minimum inhibitory concentration (MIC) values better than cloxacillin. Plumericin potently inhibited proliferation of two leukemic cancer cell lines: they were acute and chronic leukemic cancer cell lines, NB4 and K562, with the effective doses (ED50) of 4.35 ± 0.21 and 5.58 ± 0.35 μg/mL, respectively. In addition, the mechanism of growth inhibition in both cell lines was induced by apoptosis, together with G2/M arrest in K562 cells.

Antiherpetic Effects of Gynura procumbens.

The ethanol extract of Gynura procumbens showed virucidal and antireplicative actions against herpes simplex virus HSV-1 and HSV-2. It was further chromatographed on MCI gel CHP20P column giving the extract fractions F1 (water), F2 (water-methanol) F3 (methanol), and F4 (ethyl acetate). All but F1 had virucidal action against both viral types. We reported here the active compounds from F2 and F3. The antiherpetic compounds of F2 was a mixture of dicaffeoylquinic acids with virucidal and antireplicative actions against HSV-2 (IC50 96.0 and 61.0 μg/mL, resp.) Virucidal compounds of F3 were a mixture of β-sitosterol and stigmasterol (IC50 250.0 μg/mL against HSV-1), a mixture of β-sitosteryl and stigmasteryl glucosides (IC50 50.0 μg/mL against HSV-2) and 1, 2-bis-dodecanoyl-3-α-D-glucopyranosyl-sn-glycerol (IC50 of 40.0 μg/mL against HSV-2). Herbal products containing 1 and 2% of standardized ethanol extract were prepared. Double-blind randomized controlled clinical trial of the products was performed in patients with recurrent herpes labialis. Results showed that the number of patients, whose lesions healed within 7 days and the average healing time of both groups differed insignificantly. Viral culture on D7 indicated a decrease of infected patients from 48.7% to 7.69% in treated group whereas in placebo group the infected patients decreased from 31.25% to 20.00%. The viral reduction in treated group indicated the benefit of the product. Insignificant result might arise from a low number of participated patients and insufficient concentration of plant extract in herbal product.

Antiproliferative Effect and the Isolated Compounds of Pouzolzia indica.

Previous report showed the high potent antiproliferative effect of the methanolic part extracted from the aerial parts of Pouzolzia indica on NB4 and HT93A acute leukemic cell lines with the IC50 values of 28.5 and 49.8 μg/mL, respectively. The bioassay-guided fractionation of the methanolic part gave 5 fractions, that is, FFI–FFV. FFII, FFIII, and FFIV inhibited the above leukemic cell lines with the IC50 values of 15.1 (FFII), 14.4 (FFIII), 32.1 (FFIV), and 31.0 (FFII), 9.7 (FFIII), 10.5 (FFIV) μg/mL, respectively. The compounds in these fractions were isolated using chromatographic technique. FFII contained friedelin 1, 28-hydroxy-3-friedelanone 2, and 7-methoxy-coumarin 3. FFIII contained 6, 7-dimethoxy-coumarin 4, scopoletin 5, methyl caffeate 6. FFIV contained sitosteryl glucoside 7 and a supposed glycosphingolipid 8. The chemical structures were elucidated by spectroscopic methods.

Inhibitory effect of Phlai capsules on skin test responses among allergic rhinitis patients: a randomized, three-way crossover study

BACKGROUND Zingiber cassumunar Roxb., commonly known as Phlai in Thai, has been used as a traditional medicine in Thailand for the treatment of various diseases, including inflammation and chronic airway disease. OBJECTIVE The purpose of this study was to assess the antihistaminic effect of Phlai on skin testing. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION This was a randomized, open-label, three-way crossover study. Twenty allergic rhinitis (AR) patients were enrolled. In randomized sequence, patients received a single dose of Phlai capsules (100 or 200 mg) or loratadine (10 mg) with a washout period of 1 week between each treatment. MAIN OUTCOME MEASURES Skin prick testing for histamine and common aeroallergen (house dust mite) were performed before treatment and after 1, 2, 3, 4, 6, 8, 12 and 24 hours of treatment. The main treatment outcomes were the mean wheal and flare responses to the skin prick test after treatment. RESULTS Both 100 mg and 200 mg Phlai doses suppressed wheal and flare responses to house dust mite allergen, but only 200 mg of Phlai capsules significantly suppressed wheal and flare responses to histamine. Repeated measures analysis of variance showed that loratadine caused more wheal and flare suppression than Phlai capsules in responses to the histamine skin prick test. However, there were no significant differences among the effects of 100 mg Phlai capsules, 200 mg Phlai capsules and loratadine in suppression of wheal and flare induced by the mite skin prick test. Both doses of Phlai were well-tolerated with no adverse events. CONCLUSION Both 100 mg (compound D 4 mg) and 200 mg (compound D 8 mg) Phlai capsules, when taken as a single therapeutic dose, inhibited skin reactivity to histamine and mite skin prick tests in AR patients. TRIAL REGISTRATION Thai clinical trial registry (TCTR20160510001).

Botanicals in Dietary Supplements

Botanicals are accepted worldwide as medicinal agents and nutraceuticals. Extensive scientific investigations have been performed over the past 200 years which have resulted in the evolution of botanical utilization. This special issue highlights some of this research in 7 review articles and in 24 original research articles. Review Articles. W. Jiratchariyakul and G. B. Mahady wrote about the botanical status and popular herbs in EU, US, and Thailand. M. Miroddi et al. updated the market and the regulatory of botanical products in EU and US. Traditional Chinese and Indian Medicines (TCM and TIM) play an important role in Asian countries. Z. Wang et al. presented and discussed the role of TCM in the treatment of epidemic type II diabetes mellitus. M. M. Pandey et al. reported the use of TIM as a nutritional supplement in malnutrition. Y. Kamisah et al. presented the chemoprevention and antioxidation of Parkia speciosa. A. P. Bartolome et al. reviewed laboratory evidence of Bidens pilosa. T.-P. Huynh et al. discussed the promising botanical compounds for prevention and treatment of eye diseases. Research Articles. New biological and pharmacological activities of botanicals are reported. They included the enhancement of learning, memory and antistress of Acanthopanax trifoliatus (P. Sithisarn et al.), the sedative effect of Ziziphus mauritiana (A. M. M. San et al.), the cardioprotective effect of Phyllanthus emblica (L. Chularojmontri et al.), the cytoprotection and antioxidative stress of Citrus maxima (L. Chularojmontri et al.), anti-influenza viral activity of Momordica charantia (V. Pongthanapisith et al.), antioxidation from Nypa fruticans (N. Prasad et al.), Nigella glandulifera (J. Zhao et al.), TIM (M. M. Pandey et al.), Herba Cynomorii (J. Chen et al.). The alleviation of metabolic disorder of Citrus ichangensis (X. Ding et al.), and anticancer activity of Vitex agnus-castus (S. Li et al.). The mechanisms of anticancer action are deeply investigated with the botanical compounds, zerumbone from Zingiber zerumbet (N. M. Nadzri et al.), phenyl butenoid dimmer from Zingiber cassumunar (T. Anasamy et al.), and girinimbine from Murraya koenigii (S. Mohan et al.). Biochanin A, the major isoflavone from Trifolium pratense, prevented the bone loss in the ovariectomized rat (S.-J. Su et al.). Quercetin isolated from Caesalpinia mimosoides had neuroprotective effect (N. Tangsaengvit et al.) and inhibited eosinophile (M. K. Asano Sakai-Kashiwahara). C. Li et al. reported new anti-inflammatory triterpenoids from Illicium difengpi. Besides the herbal activities, the cohort study of TCM, Si-Wu-Tang, in postpartum women was performed to evaluate the health benefits. The effectiveness of the modern herbal drug was carried out using the double-blind randomized controlled clinical trial, as shown under the title “Antiherpetic effects of Gynura procumbens” (S. Jarikasem et al.). The research on the quality assessment of the botanicals was also presented in this issue. It included the HPLC analysis of Moringa oleifera (B. Vongsak et al.) and Pueraria tuberosa (S. Rastogi et al.). In addition, the arsenic accumulation in Zingiberaceous rhizomes was reported (C. Ubonnuch et al.). Weena Jiratchariyakul Ludger Beerhues Gail B. Mahady Tanawan Kummalue Molvibha Vongsakul

Experimental Therapeutics in Breast Cancer Cells

Many cancer patients have medicated themselves with traditional herbal remedies, which are mostly composed of a single or combined medicinal plants. The effectiveness of the remedies can be explained by the presence of the phytochemicals and the bioactivity. The investigators select the traditional-used and evidence-based herbs for the research and development. The investigation of the following Thai herbs and their reasonable utilization are presented.