Study of the safety of oral Triphala aqueous extract on healthy volunteers.

BACKGROUND: Triphala extract is a well known medicinal herbal formula which is usually prescribed by Thai traditional doctors to adjust the physiological functions of the body. Previous studies have reported that Triphala has antioxidant, anti-inflammatory, antihypercholesterolemia and anticancer properties. Though this herbal recipe is commonly used in Thailand, its human safety, especially in the oral form, has not been studied. We therefore conducted a clinical trial (Phase I). OBJECTIVE: This study evaluated the safety of administering the aqueous extract of Triphala to healthy volunteers at 2500 mg/d. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: An open-label, single-arm trial was conducted at Chulabhorn International College of Medicine, Thammasat University, Pathum Thani, Thailand, between July 2017 and July 2018. The study enrolled 10 male and 10 female healthy volunteers; all were given Triphala (water extract; five capsules of 500 mg each) orally, once a day, at bedtime, for four consecutive weeks. MAIN OUTCOME MEASURES: Signs and symptoms, physical examinations, hematology and blood chemistry were assessed at the beginning of the trial and every week thereafter, for four consecutive weeks. After finishing the trial, on day 28, all volunteers were invited to a follow-up session on day 35 to evaluate the safety of the herbal recipe using the same measurements. RESULTS: At the oral dose of 2500 mg/d, Triphala had no serious adverse effects in healthy volunteers. Moreover, it was found to have significantly improved the volunteers' high-density lipoprotein cholesterol (HDL-C) levels on day 35 and also reduced their blood sugar levels on days 14 and 35. CONCLUSIONS: We conclude that aqueous extract of Triphala is safe for healthy volunteers and that it elevates HDL-C levels and lowers blood sugar. Further clinical study should investigate its effects on HDL-C and blood sugar levels among the dyslipidemic and prediabetic groups. TRIAL REGISTRATION: This trial was registered in the Thai Clinical Trial Registry with the identifier TCTR20180423002.

Pharmacokinetics of cucurbitacin B from Trichosanthes cucumerina L. in rats.

BACKGROUND: Cucurbitacin B is the major bioactive constituent in Trichosanthes cucumerina L. fruits, which the pharmacological properties have been studied for decades particularly an anti-tumor activity. The pharmacokinetic profile of this compound is still limited and investigation is needed for further phytopharmaceutical product development. This study aimed to investigate the pharmacokinetic profile of cucurbitacin B after administering the compound at different doses and routes to rats. METHODS: Male Wistar rats (n = 6) were treated by cucurbitacin B extracted from Trichosanthes cucumerina L. The cucurbitacin B was administered at 0.1 mg/kg intravenously or by oral gavage at 2-4 mg/kg. Blood samples and internal organs were collected serially within 24 h after administration. Urine and feces were collected from time 0 to 48 h. The level of cucurbitacin B in biological samples was determined by liquid chromatography-tandem mass spectrometry. RESULTS: The absolute oral bioavailability of cucurbitacin B was approximately 10%. The maximum concentration in plasma after normalization by dose ranged from 4.85-7.81 µg/L and the time to reach maximum value was approximately within 30 min after oral dosing. The level of cucurbitacin B in plasma increased proportionally to the given dose. After intravenous administration, cucurbitacin B had a large volume of distribution of about 51.65 L/kg and exhibited a high tissue to plasma concentration ratio, approximately 60 to 280-fold in several organs. Negligible amount of unchanged cucurbitacin B could be detected in urine and feces and accounted less than 1% of administered dose. CONCLUSION: Cucurbitacin B had low oral bioavailability, but could be distributed extensively into internal organs with a high volume of distribution and tissue to plasma ratio. Only negligible amounts of unchanged cucurbitacin B were excreted via urine and feces suggesting that the compound might be biotransformed before undergoing an excretion. Further studies of the metabolic pathway and tissue uptake mechanism are required to strategize the future development of cucurbitacin B into clinical studies.

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.

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 µ g/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) µ g/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.

Significant increase in cytotoxic T lymphocytes and natural killer cells by triphala: a clinical phase I study.

Background. Searching for drugs or herbal formulations to improve the immunity of HIV/AIDS positive people is an important issue for researchers in this field. Triphala, a Thai herbal formulation, is reported to have immunomodulatory effects in mice. However, it has not yet been investigated for immunostimulatory and side effects in healthy human volunteers. Objective. To evaluate the immunostimulatory and side effects of Triphala in a clinical phase I study. Materials and Methods. All volunteers took Triphala, 3 capsules per day for 2 weeks. Complete physical examination, routine laboratory analysis, and immunological studies were performed before ingestion and after initial meeting for 4 consecutive weeks. Results. We found that Triphala demonstrated significant immunostimulatory effects on cytotoxic T cells (CD3(-)CD8(+)) and natural killer cells (CD16(+)CD56(+)). Both of them increased significantly when compared with those of the control samples. However, no significant change in cytokine secretion was detected. All volunteers were healthy and showed no adverse effects throughout the duration of the study. Conclusion. Triphala has significant immunostimulatory effects on cellular immune response, especially cytotoxic T cells and natural killer cells. Increases in the absolute number of these cells may provide a novel adjuvant therapy for HIV/AIDS positive people in terms of immunological improvement.

Comparison of Pouzolzia indica methanolic extract and Virkon against cysts of Acanthamoeba spp.

The present study was conducted to investigate the morphological and structural changes of Acanthamoeba cysts after being treated with various concentrations of Pouzolzia indica methanolic extract fraction 3 (methanol eluted) and Virkon solution. Changes in the Acanthamoeba cysts were detected by light microscopy, scanning electron microscopy and transmission electron microscopy. The results show Acanthamoeba cysts were killed by Pouzolzia indica methanolic extract fraction 3 at a concentration of 1:8 and by Virkon solution at a concentration of 0.25%, with a minimal cysticidal concentration (MCC) by 24 hours. Both agents caused similar structural damage to Acanthamoeba cysts in the same sequence. Step by step structural alterations occurred within the cyst. First, the cyst shrank, collapsed and had clumping of cytoplasmic stuctures inside the cyst walls. Second, the cysts began to bulge, swell, have a decrease in wrinkles in the cyst walls and spill the cytoplasmic contents into the environment. Finally, the cyst walls broke into small pieces. This study may be beneficial to compare with future studies of pharmaceutical agents against Acanthamoeba keratitis.

Pouzolzia indica methanolic extract fraction 2 and povidone-iodine induced changes in the cyst of Acanthamoeba spp.: light and electron microscopic studies.

OBJECTIVE: To compare the minimal cysticidal concentration (MCC) between Pouzolzia indica methanolic extract fraction 2 and Povidone-lodine (PVP-I) on the Acanthamoeba cyst and to illustrate the morphological changes of the cyst after being treated by light and electron microscopies. MATERIAL AND METHOD: Acanthamoeba spp were isolated from patients with Acanthamoeba keratitis and cultured on a non-nutrient agar plate (NNA) seeded with heat killed Escherichia coli (NNA-E.coli) at 37 degrees C for 7 days, adjusted to a final concentration of 10(4) cysts/ml. Several concentrations of PVP-I and fraction 2 of Pouzolzia indica methanolic extract were tested to find the minimal cysticidal concentrations (MCC) of both agents, at these concentrations there was no viable cyst which was confirmed by no excystment after further incubation for 7 days. The cysts were prepared for routine transmission and scanning electron microscopic studies. RESULTS: Structural damages of the treated cysts by MCC of PVP-I and fraction 2 of Pouzolzia indica methanolic extract showed a series of damages. Starting from shrinkage, destruction or rupture of the cyst walls and opercula, withdrawal of the cytoplasm or edema cyst by the outside solution passed through the damaged wall caused a decrease in wrinkle ridges of the ectocyst. Then the cyst was ripped and torn into small pieces CONCLUSION: MCC of PVP-I solution and the fraction 2 of Pouzolzia indica methanolic extract were 0.04% and 1:4, respectively. The structural damages were somewhat similar, such as the shrinkage, ruptured cyst wall and opercula, edema and end by breaking up of the cyst wall and degeneration of the inside cytosol. Pouzolzia indica may be modified as an effective disinfectant solution for a contact lens case if the active ingredients are more purified.

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 effect of Erycibe elliptilimba on human breast cancer cell lines.

Erycibe elliptilimba Merr. & Chun., family Convolvulaceae, is a Thai traditional medicine which has long been prescribed for various infectious and malignant diseases. Bio-assays of extracts from Erycibe elliptilimba Merr. & Chun. showed that a fraction (fraction 3) from an methanolic extract had an antiproliferative effect on SKBR3 and MDA-MB435 human breast cancer cells. The ED50 value of Erycibe elliptilimba Merr. & Chun. fraction 3 was 56.07 and 30.61 microg/ml for SKBR3 and MDA-MB435, respectively. After 48 h of exposure, this fraction at a concentration of 100 microg/ml significantly reduced cell proliferation in both cancer cells. In MDA-MB435 cells, cell cycle analysis showed that the herb extract fraction 3 induced the accumulation of cells in G2/M phase, whereas no significant change in cell cycle was detected in SKBR3 cells. The results indicated that the extract fraction 3 could induce cell cycle arrest in some way. However, further investigation is needed to assess the molecular mechanisms mediated anticancer activities of this plant.

Antibacterial activities of four Thai medicinal plants.

Medicinal plants have long been used and prescribed in Thailand for centuries. Some of them have been used for treating various diseases including infectious diseases. Pouzolzia pentandra Benn., Gelonium multiflorum A. Juss., Erycibe elliptilimba Merr. and Chun., Balanophora abbreviate Bl. are Thai medicinal plants from the Thai pharmacopoeia that have been prescribed for treating unknown fevers including some specific infectious diseases. This investigation demonstrated the effects of these Thai medicinal plants for their antibacterial activities by using the macrodilution assay. Based on the present study, the water methanol fraction (fraction 2) of Balanophora abbreviate Bl. showed the antibacterial activity at the MIC level of 250 microg/ml but the activity was bacteriostatic in its effects. Therefore, the use of these medicinal plants in controlling fever and infectious diseases appears to be justified and further investigations may be required to obtain more information.

Molecular mechanism of herbs in human lung cancer cells.

Herbs have been considered natural and valuable sources for anticancer drug discovery. Herbal medicine has been prescribed in many countries over centuries for treating various diseases including infectious and malignant diseases. Nowadays, many of the drugs that have been used for treatment of malignant diseases are derived from natural products such as Taxol, a natural product isolated initially from Pacific Yew (Taxus brevifolia). This review article describes research on molecular mechanisms, especially cytotoxic effect of natural products from plant sources, primarily preclinical studies, involving human lung cancer cells in vitro for providing more knowledge and issues for potential drug development from medicinal herbs in the future.