Phyto-progestins for the treatment of abnormal uterine bleeding without organic cause in women at high risk for breast cancer and breast cancer survivors: a prospective, pilot study.

OBJECTIVES: Some plants, such as Dioscorea Villosa (DV), Vitex Agnus Castus (VAC) and Turnera diffusa (D) have some 'progesterone-like' properties. We have investigated their simultaneous administration in breast cancer (BC) survivors or carriers of specific genetic mutations that can increase the risk of developing BC suffering from abnormal uterine bleeding without organic cause. METHODS: Women with irregular cycles in terms of length (interval between ≤ 24 or ≥ 38 days) without a uterine organic disease (polyps, adenomyosis, fibroids, hyperplasia/malignancy) were included. A daily diary of bleeding, questionnaires about health-related quality of life (Short Form 36) and menstrual psychophysical well-being (PGWB-1) and the Greene Climacteric Scale (GCS) (in women older than 40 years old) questionnaire were used. The presence of some premenstrual syndrome (PMS) symptoms was also evaluated. RESULTS: In the analyzed group of women (n = 15), all experienced a regularization of the menstrual cycles, with a mean duration in the three months of use of 27.1 ± 3.2 days, with a significant reduction of menstrual pain (p = 0.02) and flow (p = 0.02) intensity. Women with PMS (7/15) reported an impovement in depression, headache and abdominal pain scores (p < 0.05). No specific deterioration of different questionnaires evaluated during treatment were observed. General satisfaction with the treatment was 6.8 ± 0.3/10 on a 10 point. CONCLUSIONS: A combination of DV, VAC and D could be a promising candidate to treat menstrual irregularities without an organic cause, with a significant reduction of menstrual pain and flow intensity and possible additional benefits in PMS symptoms treatment in women at genetic risk for BC and BC survivors.

Trichilia catigua and Turnera diffusa phyto-phospholipid nanostructures: Physicochemical characterization and bioactivity in cellular models of induced neuroinflammation and neurotoxicity.

Flavonoid-based therapies supported by nanotechnology are considered valuable strategies to prevent or delay age-related and chronic neurodegenerative disorders. Egg yolk phospholipids were combined with flavonoid-rich extracts obtained from Trichilia catigua A.Juss. (rich in flavan-3-ols and phenylpropanoid derivatives) or Turnera diffusa Willd. ex Schult (dominated by luteolin derivatives) to prepare nanophytosomes. The nanophytosomes showed that size and surface charge of the lipid-based vesicles are dependent of their phenolic composition. In vitro assays with SH-SY5Y cells showed that both formulations protect cells from glutamate-induced toxicity, but not from 6-hydroxydopamine/ascorbic acid. T. diffusa nanophytosomes promote a decrease of nitric oxide produced by BV-2 cells stimulated with interferon-γ. Nanophytosomes dialysed against a mannitol solution, and then lyophilised, allow to obtain freeze-dried products that after re-hydration preserve the essential physicochemical features of the original formulations, and exhibit improved colloidal stability. These results indicate that these flavonoid/phospholipid-based nanophytosomes have suitable features to be considered as tool in the development of therapeutic and food applications.

Trichilia catigua and Turnera diffusa extracts: In vitro inhibition of tyrosinase, antiglycation activity and effects on enzymes and pathways engaged in the neuroinflammatory process.

ETHNOPHARMACOLOGICAL RELEVANCE: Flavonoids interact with multiple targets in Central Nervous System resulting in a broad neuroprotection mediated by complementary processes and synergic interactions. Therefore, flavonoid-based therapies may input positive outcomes in the prevention and early management of neurodegenerative diseases. In Brazilian folk medicine Trichilia catigua is used for its neuroactive properties, such as neurostimulant, antioxidant and anti-neuroinflammatory, while Turnera diffusa is traditionally used as a tonic in neurasthenia. Both species are known to be rich in flavonoids. AIM OF THE STUDY: To study aqueous extracts of T. catigua and T. diffusa in terms of their antioxidant and antiglycation effects, inhibition of tyrosinase activity, and interaction with enzymes and pathways engaged in neuroinflammation. Moreover, whenever possible, to establish a relationship between the studied activities and the traditional usage of the species. MATERIALS AND METHODS: The phenolic profiles of the aqueous extracts were validated by HPLC-DAD. The effect of the extracts over mushroom tyrosinase and 5-lipoxygenase activities, as well as their capacity to impair bovine serum albumin glycation, were assessed by in vitro assays. The anti-neuroinflammatory potential of the same extracts was evaluated by their capacity to mitigate the pro-inflammatory stimulus induced in BV-2 microglia cells by interferon-gamma. RESULTS: T. catigua extract, a rich mixture of phenolic acids, catechins and flavonolignans, excels by its ability to decrease lipid peroxidation (EC50 = 227.18 ±â€¯9.04 µg/mL), and to work as anti-glycation agent, and inhibitor of both tyrosinase and 5-lipoxigenase (IC50 = 358.84 ±â€¯19.05 and 56.25 ±â€¯14.53 µg/mL, respectively). However, only T. diffusa extract, mainly composed by luteolin derivatives, is able to lower NO production by BV-2 microglia cells stimulated with interferon-gamma, despite its lower activities in the other assays. CONCLUSIONS: Overall, this work highlights the value of medicinal plant extracts as sources of bioactive flavonoid-rich extracts with neuroactive effects. Furthermore, these results support their application as alternative strategies to develop functional foods and therapeutics to fight chronic neurodegenerative disorders.

Turnera diffusa extract attenuates profibrotic, extracellular matrix and mitochondrial markers in activated human hepatic stellate cells (HSC).

INTRODUCTION AND OBJECTIVES: Hepatic fibrosis is characterized by the accumulation of extracellular matrix which includes the accumulation of α-smooth muscle actin (α-SMA), collagen type I (COL1α1), as well as remodeling induced by metalloproteinases and tissue inhibitor of metalloproteinase (TIMPs), where hepatic stellate cells (HSCs) play a central role. In addition, the transcription factor SNAI1 (which participates in epithelial-mesenchymal transition, EMT) and mitofusin 2 (MFN2, a mitochondrial marker) plays an important role in chronic liver disease. Turnera diffusa (TD), a Mexican endemic plant, has been shown to possess antioxidant and hepatoprotective activity in vitro. We treated human HSC (LX2 cells) with a methanolic extract of Turnera diffusa (METD) to evaluate the mechanism involved in its hepatoprotective effect measured as fibrosis modulation, EMT, and mitochondrial markers. MATERIALS AND METHODS: HSC LX-2 cells were treated with METD (100 and 200ng/mL) alone or combined with TGF-ß (10ng/mL) at different time points (24, 48, and 72h). α-SMA, COL1α1, MMP2, TIMP1, SNAI1, and MFN2 mRNAs and protein levels were determined by real-time quantitative PCR and Western Blot analysis. RESULTS: We found that METD decreases COL1α1-mRNA, α-SMA, and TIMP1 protein expression in LX2 cells treated with and TGF-ß. This treatment also decreases MFN2 and TIMP1 protein expression and induces overexpression of MMP2-mRNA. CONCLUSIONS: Our results suggest that a methanolic extract of Turnera diffusa is associated with an antifibrotic effect by decreasing profibrotic and mitochondrial markers together with the possible induction of apoptosis through SNAI1 expression in activated HSC cells.

Evaluation of the testicular protection conferred by damiana (Turnera diffusa Willd.) against amitriptyline-induced testicular toxicity, DNA damage and apoptosis in rats.

Psychiatric drugs, such as antidepressants, are used to treat depression based on their ability to modify chemical imbalances of the key neurotransmitters in the brain, including dopamine, serotonin, and norepinephrine. Amitriptyline, a first-reference tricyclic antidepressant derived from dibenzocycloheptadine, is frequently used, especially in neuropsychiatry, to address general depression, major depressive disorders, and fibromyalgia. Therefore, this study attempted to examine the sexual dysfunction attendant on the use of Amitriptyline by investigating the protective role that can be played by damiana. To this end, this study used damiana (Turnera diffusa Willd.) as adjuvant therapy in male albino rats receiving Amitriptyline. Sixty male albino rats were randomly allocated to six groups, with 10 rats being assigned to each group; the first group was a control, the second was treated with damiana only, the third group was given Amitriptyline, the fourth group received Amitriptyline and damiana simultaneously, the fifth group was given Amitriptyline and post-treated with damiana, and the sixth group was given Amitriptyline and then allowed time for self-healing. The findings of this study suggest that oxidative stress occurs in testicular tissue in rat groups treated with Amitriptyline, as manifested by inappropriate activity of TBARS, SOD, GSH, GR, GST, and GPx. Amitriptyline also repressed reproductive hormonal activity, as confirmed by histopathological lesions, DNA damage, and p53 protein expression. The addition of damiana, however, showed aprotective role in all testicular activity indices.

Selective Inhibition of Human Monoamine Oxidase B by Acacetin 7-Methyl Ether Isolated from Turnera diffusa (Damiana).

The investigation of the constituents that were isolated from Turnera diffusa (damiana) for their inhibitory activities against recombinant human monoamine oxidases (MAO-A and MAO-B) in vitro identified acacetin 7-methyl ether as a potent selective inhibitor of MAO-B (IC50 = 198 nM). Acacetin 7-methyl ether (also known as 5-hydroxy-4', 7-dimethoxyflavone) is a naturally occurring flavone that is present in many plants and vegetables. Acacetin 7-methyl ether was four-fold less potent as an inhibitor of MAO-B when compared to acacetin (IC50 = 50 nM). However, acacetin 7-methyl ether was >500-fold selective against MAO-B over MAO-A as compared to only two-fold selectivity shown by acacetin. Even though the IC50 for inhibition of MAO-B by acacetin 7-methyl ether was ~four-fold higher than that of the standard drug deprenyl (i.e., SelegilineTM or ZelaparTM, a selective MAO-B inhibitor), acacetin 7-methyl ether's selectivity for MAO-B over MAO-A inhibition was greater than that of deprenyl (>500- vs. 450-fold). The binding of acacetin 7-methyl ether to MAO-B was reversible and time-independent, as revealed by enzyme-inhibitor complex equilibrium dialysis assays. The investigation on the enzyme inhibition-kinetics analysis with varying concentrations of acacetin 7-methyl ether and the substrate (kynuramine) suggested a competitive mechanism of inhibition of MAO-B by acacetin 7-methyl ether with Ki value of 45 nM. The docking scores and binding-free energies of acacetin 7-methyl ether to the X-ray crystal structures of MAO-A and MAO-B confirmed the selectivity of binding of this molecule to MAO-B over MAO-A. In addition, molecular dynamics results also revealed that acacetin 7-methyl ether formed a stable and strong complex with MAO-B. The selective inhibition of MAO-B suggests further investigations on acacetin 7-methyl as a potential new drug lead for the treatment of neurodegenerative disorders, including Parkinson's disease.

Modulatory effect of Turnera diffusa against testicular toxicity induced by fenitrothion and/or hexavalent chromium in rats.

Oxidative stress and increased production of reactive oxygen species have been implicated in pesticides and heavy metals toxicity. The objective of this study was to investigate the efficacy of Turnera diffusa Willd (damiana) on counteracting fenitrothion (FNT) and/or potassium dichromate (CrVI)-induced testicular toxicity and oxidative injury in rats. FNT and/or CrVI intoxicated animals revealed a significant increase in thiobarbituric acid reactive substances and hydrogen peroxide levels. While, reduced glutathione and protein content, as well as antioxidant enzymes, phosphatases, and aminotransferases activities, were significantly decreased. In addition, significant changes in testosterone and follicle-stimulating hormone levels were detected. Furthermore, histological and immunohistochemical alterations were observed in rat testes and this supported the observed biochemical changes. On the other hand, rats treated with damiana alone decreased lipid peroxidation and increased most of the examined parameters. Moreover, damiana pretreatment to FNT and/or CrVI-intoxicated rats showed significant improvement in lipid peroxidation, enzyme activities, and hormones as compared with their respective treated groups. Conclusively, rats treated with both FNT and/or CrVI showed pronounced hazardous effect especially in their combination group in addition, Turnera diffusa had a potential protective role against FNT and/or CrVI induced testicular toxicity.

Antioxidant effects of damiana (Turnera diffusa Willd. ex Schult.) in kidney mitochondria from streptozotocin-diabetic rats.

The antioxidant effects of water-ethanol extract (WEE) from Turnera diffusa (damiana) in kidney mitochondria from experimental streptozotocin-induced diabetes mellitus (STZ-DM) rats was evaluated. STZ-DM rats were orally treated during three and five weeks. After experimental periods, kidney mitochondria were isolated and malondialdehyde (MDA), nitric oxide (NO•) and protein nitrosylation levels were measured. Also, blood glucose (BG) and body weight (BW) were recorded. Damiana significantly reduced the MDA and NO• levels in kidney mitochondria, although no changes in protein nitrosylation were observed and it did not have the potential to reverse the hyperglycaemia. In conclusion, WEE of T. diffusa have antioxidant properties that may prevent damage induced by mitochondrial oxidative stress in kidneys of STZ-DM rats.

Plantas silvestres consumidas como tés recreativos por grupos de rancheros en Baja California Sur, México / Wild plants consumed as recreational teas by groups of ranchers in Baja California Sur, Mexico

Antecedentes y Objetivos: Los tés recreativos son bebidas preparadas, por infusión/ebullición, de diferentes partes de plantas que se consumen en un entorno social/alimenticio. Se consideran generalmente saludables y con usos medicinales, aunque ese no es su único propósito de consumo. En México, los tés recreativos prácticamente no se mencionan en los estudios etnobotánicos. Este estudio describe la etnobotánica, contenido de fenólicos y actividad antioxidante de plantas silvestres empleadas como tés recreativos en dos rancherías de las zonas montañosas de la región de Los Cabos, Baja California Sur, México: Reserva de la Biosfera Sierra La Laguna y comunidad San Blas-Cacachilas. Métodos: La información etnobotánica se obtuvo mediante encuestas semi-estructuradas y estancias con los rancheros de ambas comunidades. Se calculó el porcentaje de citas, preferencia y valor cultural de cada especie registrada, además de describir sus usos medicinales adicionales. El contenido de fenólicos y actividad antioxidante se determinó por métodos colorimétricos y fueron correlacionados con parámetros etnobotánicos. Resultados clave: Se registraron 12 taxa de plantas silvestres para elaborar tés recreativos, 11 en Sierra La Laguna y seis en San Blas-Las Cacachilas. La damiana (Turnera diffusa) obtuvo el mayor valor cultural (71.2%), seguido de margarita (Aloysia barbata) (41.1%) y anís silvestre (Tagetes filifolia/T. micrantha) (21.5%). Los tés recreativos tuvieron sus principales usos medicinales como antigripales/antitusivos y relajantes generales. El contenido de antioxidantes presentó correlación negativa con el porcentaje de citas y la preferencia. Los usos etnofarmacológicos de A. barbata y el helecho peyote (Pellaea ternifolia) son descritos por primera vez, y se documentaron nuevos usos para la damiana y el cerezo (Prunus serotina). Conclusiones: Los rancheros de Baja California Sur tienen gran afinidad por el consumo de tés recreativos de plantas silvestres, y aquellos usados para tratar padecimientos comunes como gripe/tos o estrés son los preferidos. El presente es el primer estudio etnobotánico específico para tés recreativos en México.

Medicinal species as MTDLs: Turnera diffusa Willd. Ex Schult inhibits CNS enzymes and delays glutamate excitotoxicity in SH-SY5Y cells via oxidative damage.

One of the most promising approaches to confront the complexity of central nervous system disorders are new multi-target directed ligands (MTDLs). Five medicinal species (Cereus grandiflorus (L.) Mill., Hyssopus officinalis L., Acorus calamus L., Silybum marianum L. Gaertn. and Turnera diffusa Willd. Ex Schult), selected for their ethnopharmacological relevance, were object for in vitro screening. The aqueous extract of T. diffusa revealed the strongest neuroactive potential, inhibiting monoamine oxidase-A (IC50 = 129.80 ± 11.97 µg/mL), and acetyl- and butyrylcholinesterase (IC25 = 0.352 ± 0.011 and 0.370 ± 0.036 mg/mL, respectively). Its phenolic profile was established for the first time by HPLC-DAD-ESI/MSn. Twenty-six out of thirty-seven compounds were newly identified in this species. The pre-treatment with this flavonoid-rich extract promoted a rightward shift of the glutamate concentration neuronal cell (SH-SY5Y) death response curve. Furthermore, it significantly reduced the early phase formation of intracellular reactive species after glutamate and t-BHP exposure, suggesting that neuroprotection in SH-SY5Y cells was, in part, mediated by antioxidant mechanisms.

Acute Hypoglycemic and Antidiabetic Effect of Teuhetenone A Isolated from Turnera diffusa.

Diabetes mellitus is a chronic degenerative disease that causes long-term complications and represents a serious public health problem. Turnera diffusa (damiana) is a shrub that grows throughout Mexico and is traditionally used for many illnesses including diabetes. Although a large number of plant metabolites are known, there are no reports indicating which of these are responsible for this activity, and this identification was the objective of the present work. Through bioassay-guided fractionation of a methanolic extract obtained from the aerial part of T. diffusa, teuhetenone A was isolated and identified as the main metabolite responsible for the plant's hypoglycemic activity. Alpha-glucosidase inhibitory activity and cytotoxicity of this metabolite were determined. Hypoglycemic and antidiabetic activities were evaluated in a murine model of diabetes in vivo, by monitoring glucose levels for six hours and comparing them with levels after administering various controls. Teuhetenone A was not cytotoxic at the tested concentrations, and did not show inhibitory activity in the glucosidase test, and the in vivo assays showed a gradual reduction in glucose levels in normoglycemic and diabetic mice. Considering these results, we suggest that teuhetenone A has potential as an antidiabetic compound, which could be further submitted to preclinical assays.