Paraxanthine Supplementation Increases Muscle Mass, Strength, and Endurance in Mice.

Paraxanthine is a natural dietary ingredient and the main metabolite of caffeine in humans. Compared to caffeine, paraxanthine exhibits lower toxicity, lesser anxiogenic properties, stronger locomotor activating effects, greater wake promoting properties, and stronger dopaminergic effects. The purpose of this study was to evaluate the potential beneficial effects of paraxanthine supplementation on muscle mass, strength, and endurance performance in comparison to the control and other ingredients commonly used by athletes: L-theanine, alpha-GPC, and taurine. Male Swiss Albino mice from five groups (n = 8 per group) were orally administered paraxanthine (20.5 mg/kg/day, human equivalence dose (HED) 100 mg), L-theanine (10.28 mg/kg/day, HED 50 mg), alpha-GPC (41.09 mg/kg/day, HED 200 mg), taurine (102.75 mg/kg/day, HED 500 mg), or control (carboxy methyl cellulose) for 4 weeks. Exercise performance was evaluated using forelimb grip strength and treadmill endurance exercise. All animals were subject to treadmill training for 60 min 5 days per week. Blood draws were utilized to analyze lipid profile, liver health, renal function, and nitric oxide levels. Paraxanthine significantly increased forelimb grip strength by 17% (p < 0.001), treadmill exercise performance by 39% (p < 0.001), gastrocnemius and soleus muscle mass by 14% and 41% respectively (both p < 0.001), and nitric oxide levels by 100% compared to control (p < 0.001), while reducing triglyceride (p < 0.001), total cholesterol (p < 0.001), LDL (p < 0.05), and increasing HDL (p < 0.001) compared to control, and compared to L-theanine, alpha-GPC, and taurine. Results from this initial investigation indicate that, when compared to the control, L-theanine, alpha-GPC, and taurine, paraxanthine is an effective ingredient for various aspects of sports performance and may enhance cardiovascular health.

Modulatory Effect of Carotenoid Supplement Constituting Lutein and Zeaxanthin (10:1) on Anti-oxidant Enzymes and Macular Pigments Level in Rats.

BACKGROUND: Human eye is constantly exposed to different wavelengths and intensities of light. Oxidative stress results in distinct changes to retinal organs and tissues. Macular pigments (lutein and zeaxanthin), present in the central macular region, provide protection from photodamages by absorption of high energy blue light and also by virtue of their anti-oxidant activity. Ocular phototoxicity is thus prevented by our efficient anti-oxidant system, in both young and old. One of the best commercial sources of pure lutein and zeaxanthin is Marigold flowers. OBJECTIVE: In the present study, oil-soluble dietary carotenoid supplement constituting lutein and zeaxanthin in the ratio of 10:1 was evaluated for its modulatory effect on anti-oxidant enzymes and macular pigments in the serum and macula of the Swiss albino rats. MATERIALS AND METHODS: Male Swiss albino rats were treated with carotenoid supplement constituting lutein and trans-Zeaxanthin (10:1) at two different doses daily, under standard experimental conditions for 42 days. End of the treatment, serum and macula were collected and used for measurement of lutein and zeaxanthin levels along with anti-oxidant parameters. STATISTICAL ANALYSIS USED: Statistical differences were assessed by analysis of variance (ANOVA) followed by Dunnet's test. P < 0.05 was considered statistically significant. All the results were expressed as mean ± standard deviation. RESULTS: The supplement exhibited significant elevation of anti-oxidant enzyme levels in treated animals in dose-dependent manner. Concomitantly, the total anti-oxidant capacity has also been found to show similar increment at the end of the study period. This study revealed significant expression of the two macular pigments investigated. CONCLUSIONS: Our study, therefore, provides a strong claim for the anti-oxidant effect of the oil-soluble dietary carotenoid supplement, and thus substantiates its use in the prevention of phototoxic damage to the eye on long-term supplementation. SUMMARY: Apart from its ornamental value, Marigold (Tagetes erecta L.) flowers are well known as an herbal remedy due to its antimicrobial, anti-inflammatory, and anti-oxidant activities. Epidemiological studies have implicated prolonged exposure to ultraviolet radiations & blue light and in turn oxidative stress in the pathogenesis of the majority of the eye diseases, since childhood. Studies have shown that with age a number of changes occur predisposing the retinal various organs and tissues to oxidative stress. These changes manifest in decreased levels in plasma of Vitamin C, Vitamin E, glutathione, Retinal Pigment Epithelium (RPE), Catalase (CAT), Super Oxide Dismutase (SOD), Thiobarbituric Acid Reactive Substance (TBARS), and total anti-oxidant capacity (TAC). Age- and diet-related loss of Lutein and Zeaxanthin enhance phototoxic damage to the eye, and thus supplementation of these carotenoids becomes vital for maintaining optimal eye healthIn the present study, XanMax® 2002 oil, a supplement constituting lutein and trans-Zeaxanthin, extracted from the flowers of T. erecta, was evaluated for its modulatory effect on anti-oxidant enzymes and macular pigments in the serum and macula of the Swiss albino rats. XanMax® 2002 oil exhibited significant elevation of anti-oxidant enzyme levels in treated animals in dose-dependent manner. Concomitantly, the TAC has also been found to show similar increment at the end of the study period. This study revealed significant expression of the two macular pigments investigated. Abbreviations used: AMD: Age related Macular Diseases; RPE: Retinal Pigment Epithelium; CAT: Catalase; SOD: Super Oxide Dismutase; TAC: Total Antioxidant Capacity; ROS: Reactive Oxygen Species; LC-MS: Liquid chromatography-mass spectrometry; p.o.: Per Orally; CMC Carboxymethyl cellulose.

Achyranthes aspera Attenuates epilepsy in experimental animals: possible involvement of GABAergic mechanism.

The present study was aimed to examine the possible anticonvulsant property of aerial parts of Achyranthes aspera using various experimental models of epilepsy in mice. Petroleum ether extract of aerial parts of A. aspera (PeAA), methanolic eAA (MeAA) and aqueous eAA (AeAA) was initially evaluated against six-hertz seizure model in mice, based on the outcomes the effective extract was further evaluated against maximal electroshock (MES) and pentylenetetrazole (PTZ) models in mice. In addition, the potent extract was evaluated against the PTZ model by co-administering with flumazenil (FMZ), and also evaluated for its effect on GABA levels in brain and NMDA-induced lethality in mice. Furthermore, the probable locomotor deficit-inducing property of the extract was evaluated by actophotometer test in mice. In results, only MeAA showed protection against six-hertz-induced seizures in mice, based on these outcomes only MeAA was evaluated in MES and PTZ models. Notably, the MeAA (200, 400 and 800 mg/kg) has offered mild and dose dependent protection against MES and PTZ-induced seizures in mice. Alongside, the MeAA (400 mg/kg) showed a significant increase in GABA levels in the brain compared to control, and in line with these findings the anti-PTZ effect of MeAA (400 mg/kg, p.o.) was blocked when co-administered with flumazenil (5 mg/kg, i.p.). However, the MeAA has not shown significant protection against NMDA-induced mortality and also did not cause significant change in locomotor activity compared to before treatment. These findings suggest that MeAA possess mild anticonvulsant activity and the outcomes further confirmed the involvement of GABAergic mechanism behind the anticonvulsant activity of MeAA.

Effect of Coleus forskohlii and its major constituents on cytochrome P450 induction.

Coleus forskohlii Briq. has been used traditionally for the treatment of several ailments since antiquity in Ayurveda. In the present study, an approach has been made to evaluate the effect of C. forskohlii and its major constituents on cytochrome P450 (CYP3A, CYP2B, and CYP2C) mRNA expression in rat hepatocytes. To gain better understanding of the herb-drug interaction potential of the chemical constituents present in C. forskohlii, the extract was subjected to column chromatography followed by standardization with respect to forskolin, 1-deoxyforskolin, and 1,9-dideoxyforskolin using reversed-phase high-performance liquid chromatography (RP-HPLC). Hepatocytes were treated with extracts, fractions, and phytoconstituents, followed by extraction and purification of total mRNA. Study of mRNA expression was carried out through reverse transcription polymerase chain reaction, followed by agarose gel electrophoresis. Results revealed that the test substances did not show any significant mRNA expression compared to the control against CYP3A, CYP2B, and CYP2C. Positive controls such as dexamethasone and rifampin showed significantly high (p < 0.001) induction potential compared to the control. It can be concluded that C. forskohlii and its major constituents may not be involved in CYP450 induction-based drug interaction.

Anti-hyperglycemic activity of Caralluma umbellata Haw.

INTRODUCTION: Hyperglycemia is a serious health problem prevailing in diabetes patients. Treatment for hyperglycemia by various oral anti-hyperglycemic drugs have associated with side effects, hence there is growing awareness towards the use of herbal products due to their efficacy, minimal side effects and relatively low costs. This study is designed to evaluate anti-hyperglycemic activity of Caralluma umbellata Haw, which is used as a traditional medicinal plant all over India through in vitro studies. METHODS: Methanolic, aqueous and hydro methanolic extracts of Caralluma umbellata were prepared and studied for their anti hyperglycemic activity. The extracts were evaluated for glucose uptake in L6 myotubes in vitro. In addition, the inhibitory activity against alpha amylase and pancreatic lipase was also measured. RESULTS: The methanolic extract (MCU) was found to have significant glucose uptake. Further, MCU was also found to have promising role in inhibiting alpha amylase and pancreatic lipase. CONCLUSION: The results of present study shows Caralluma umbellata has potential antidiabetic property, thus providing a further scope for study in animal model and understanding the mechanism of action.

Evaluation of antioxidant, in vitro cytotoxicity of micropropagated and naturally grown plants of Leptadenia reticulata (Retz.) Wight & Arn.-an endangered medicinal plant.

OBJECTIVE: To evaluate the antioxidant and anti proliferative potential of different solvent extract of micropropagated and naturally grown plants of Leptadenia reticulata against various cancer cell lines. METHODS: In this study different extract were tested for cytotoxicity against human breast adenocarcinoma cell line MCF-7, human colon adenocarcinoma grade II cell line HT-29 and non cancer skeletal muscle cell line L6 through 3-(4, 5-dimethyl thiazol-2-yl)-5-diphenyl tetrazolium bromide assay. The total antioxidant potential was estimated by three different antioxidant model diphenylpicrylhydrazyl free radical scavenging activity, H2O2 scavenging activity and FeCl3 reducing activity. RESULTS: The ethyl acetate extract of both naturally grown plant and tissue cultured plant exhibited significant cytotoxicity with IC50 values of 21 µg/mL, 26 µg/mL and 22 µg/mL; 20 µg/mL, 30 µg/mL and 18 µg/mL respectively against three cell lines. The diphenylpicrylhydrazyl free radical scavenging activity was found to be highest with IC50 value of 267.13 µg/mL in ethyl acetate extract. The methanolic extract exhibited moderate antioxidant activity with IC50 value of 510.15 µg/mL. A highly positive correlation was observed between the antioxidant potential and cytotoxic activity of the plant. CONCLUSIONS: The strong cytotoxicity of ethyl acetate extract revealed anti carcinogenic potential of the plant which supports its traditional use as medicine. The present investigation is new to literature till date and will provide better scientific basis for future pharmacological, in vivo studies and novel source of pure bioactive compounds having anti cancer properties in this plant.

Methanolic extract of Momordica cymbalaria enhances glucose uptake in L6 myotubes in vitro by up-regulating PPAR-γ and GLUT-4 / 中国天然药物

The present study was undertaken to evaluate the influence of the methanolic fruit extract of Momordica cymbalaria (MFMC) on PPARγ (Peroxisome Proliferator Activated Receptor gamma) and GLUT-4 (Glucose transporter-4) with respect to glucose transport. Various concentrations of MFMC ranging from 62.5 to 500 μg·mL(-1) were evaluated for glucose uptake activity in vitro using L6 myotubes, rosiglitazone was used as a reference standard. The MFMC showed significant and dose-dependent increase in glucose uptake at the tested concentrations, further, the glucose uptake activity of MFMC (500 μg·mL(-1)) was comparable with rosigilitazone. Furthermore, MFMC has shown up-regulation of GLUT-4 and PPARγ gene expressions in L6 myotubes. In addition, the MFMC when incubated along with cycloheximide (CHX), which is a protein synthesis inhibitor, has shown complete blockade of glucose uptake. This indicates that new protein synthesis is required for increased GLUT-4 translocation. In conclusion, these findings suggest that MFMC is enhancing the glucose uptake significantly and dose dependently through the enhanced expression of PPARγ and GLUT-4 in vitro.

Antioxidant and hepatoprotective activity of Fagonia schweinfurthii (Hadidi) Hadidi extract in carbon tetrachloride induced hepatotoxicity in HepG2 cell line and rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The whole plant of Fagonia schweinfurthii (Hadidi) Hadidi (Family: Zygophyllaceae) is used in variety of diseases including hepatic ailments in deserts and dry areas of India. AIM OF THE STUDY: To evaluate antioxidant and hepatoprotective activity of ethanolic extract from Fagonia schweinfurthii (Hadidi) Hadidi (FSEE) in carbon tetrachloride (CCl4) induced hepatotoxicity in HepG2 cell line and rats. MATERIALS AND METHODS: In vitro antioxidant activity was determined by DPPH, ABTS radicals and hydrogen peroxide methods. In vitro cytotoxicity and hepatoprotective potential of FSEE were evaluated using HepG2 cells. Based on the cytotoxicity assay, FSEE (50, 100, 200 µg/ml) was assessed for hepatoprotective potential against CCl4 induced toxicity in HepG2 cell line by monitoring cell viability, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH) leakage, lipid peroxidation (LPO) and glutathione level (GSH). Further, in vivo hepatoprotective activity of FSEE was evaluated against CCl4 induced hepatotoxicity in male Wistar albino rats. Rats were pre-treated with FSEE (200 mg, 400 mg kg(-1) day(-1) p.o.) for 7 days followed by a single dose of CCl4 (1.0 ml/kg, i.p.) on 8th day. Silymarin was used as positive control. After 24h of CCl4 administration, various biochemical parameters like aspartate aminotransferase (AST), alanine aminotransaminase (ALT), alkaline phosphatase (ALP), total bilirubin (TB) and total protein (TP) levels were estimated in serum. The antioxidant parameters like superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione (GSH) content and malondialdehyde (MDA) level in the liver homogenate were determined. Histopathological changes in the liver of different groups were also studied. RESULTS: The FSEE possessed strong antioxidant activity in vitro. The results indicated that CCl4 treatment caused a significant decrease in cell viability. The CCl4-induced changes in the HepG2 cells were significantly ameliorated by treatment of the FSEE. FSEE significantly prevented CCl4 induced elevation of AST, ALT, ALP, TB, and CCl4 induced decrease in total protein in rats. FSEE treated rat liver anti-oxidant parameters (SOD, CAT, MDA and GSH,) were significantly antagonized for the pro-oxidant effect of CCl4. Histopathological studies also supported the protective effect of FSEE. CONCLUSION: The results of this study revealed that FSEE has significant hepatoprotective activity. This effect may be due to the ability of the extract to inhibit lipid peroxidation and increase in the anti-oxidant enzymatic activity.

Bioactivity-guided isolation of cytotoxic constituents from three medicinal plants.

CONTEXT: The ethanol extracts and their fractions of three Indian medicinal plants, Ervatamia coronaria (Jacq.) Stapf, (Apocynaceae), Mimosa pudica L. (Mimosaceae) and Caesalpinia bonduc (L.) Roxb. (Caesalpiniaceae) were tested for their cytotoxic activity in the brine shrimp lethality (BSL) bioassay and in various cancer cell lines. The plants were selected based on their traditional use in the treatment of cancer/tumors. OBJECTIVES: To investigate the in vitro cytotoxicity of Ervatamia coronaria, Mimosa pudica and Caesalpinia bonduc. MATERIALS AND METHODS: Ethanolic extracts and their fractions of E. coronaria, M. pudica and C. bonduc were subjected to cytotoxicity studies using BSL bioassay method with concentrations of 10, 50, 100, 500 and 1000 µg/ml. The alkaloid fraction of E. coronaria with significant cytotoxicity in BSL bioassay was subjected to in vitro cytotoxicity studies with HT-29, A-549, HepG-2, MCF-7 and L-6 cell lines at concentrations of 12.5, 25, 50, 100 and 200 µg/ml and a DNA fragmentation study using the HT-29 cell line. RESULTS: The alkaloid fractions of E. coronaria and M. pudica showed significant cytotoxicity with LC50 values of 65.83 and 85.10 µg/ml in the BSL bioassay, respectively. The purified alkaloid fraction of E. coronaria exhibited highest cytotoxicity in HT-29, A-549 and MCF-7 cell lines with IC50 values of 32.5, 47.5 and 72.5 µg/ml, respectively, and induced DNA fragmentation in the HT-29 cell line at a concentration of 65 µg/ml. CONCLUSION: The alkaloid fraction of E. coronaria exhibited significant cytotoxicity. Alkaloids such as ervatamine, apparicine and coronaridine that were earlier reported may be responsible for this activity.

A novel herbal formulation "LiverCare" differentially regulates primary rat hepatocyte and hepatocarcinoma cell proliferation in vitro.

Hepatocyte growth factor (HGF) plays an important role in hepatocyte proliferation. HGF expression is regulated by various signaling molecules and nuclear receptors. In the present study, LiverCare(®) (LC), a novel polyherbal formulation (The Himalaya Drug Company, Bangalore, India), was evaluated for its efficacy, using co-cultures of primary rat hepatocytes-non-parenchymal cells (NPCs) and human hepatocellular carcinoma cells (HepG2). The rate of primary hepatocyte co-culture proliferation was significantly and dose-dependently increased by LC as determined by [(3)H]thymidine incorporation into newly synthesized DNA and cell proliferation assay. LC also increased HGF expression in primary hepatocyte co-culture. Albumin and urea content remained constant during proliferation of hepatocyte co-cultures in the presence of LC with decreased activity of alanine aminotransferase. It is interesting that LC inhibited incorporation of [(3)H]thymidine into DNA in HepG2 cells. LC enhanced peroxisome proliferator-activated receptor-α expression during hepatocyte proliferation, whereas tumor necrosis factor-α expression remained unaffected. In conclusion, our study clearly showed that LC differentially regulates primary rat hepatocytes and human hepatocarcinoma cell proliferation. LC may be a promising candidate for treating degenerative liver diseases by enhancing liver regeneration.

Water-soluble compounds in the herbal preparation Abana inhibit lipid biosynthesis and enhance cholesterol efflux in HepG2 cells.

Higher concentrations of circulating lipids (cholesterol and triglycerides) and their decreased catabolism pose a major risk in the development of atherosclerosis and coronary heart disease (CHD). Although statins are widely used for treatment of hyperlipidemia, side effects associated with their use have prompted the search for a safer alternative for treating hyperlipidemia. The present study investigated the effect of water-soluble compounds in Abana (WSCA), a polyherbal drug formulation traditionally used in India for the treatment of hyperlipidemia, on lipid metabolism in HepG2 cells. WSCA reduced cholesterol and triglyceride content in the cells and their supernatant. WSCA inhibited the incorporation of [2-14C]acetate into cellular cholesterol and fatty acids, suggesting the inhibition of lipid synthesis. In addition, WSCA inhibited HMG-CoA reductase, a key metabolic enzyme involved in the biosynthesis of cholesterol. WSCA also increased cholesterol and fatty acid secretion into the cell supernatant, suggesting the enhanced removal of cholesterol and fatty acids. Furthermore, WSCA showed decreased linoleic acid (18:2) and arachidonic acid (20:4) content in HepG2 cells. The present study is the first to show that WSCA simultaneously inhibited cellular cholesterol biosynthesis and increased cholesterol secretion into the cell supernatant in HepG2 cells.

Liv.52 regulates ethanol induced PPARgamma and TNF alpha expression in HepG2 cells.

Liver is a prime target of alcohol-induced damage by inducing inflammatory cytokines especially tumor necrosis factor alpha (TNFalpha). Activator of peroxisome proliferator activator receptor gamma (PPARgamma) is protective against alcohol-induced liver injury in animals. Liv.52, one of the major herbal hepatoprotective drugs, is shown to protect the liver from toxicity and is considered to be an effective hepatoprotective agent. However, the signal pathway involved in the Liv.52-induced hepatoprotection is not understood well especially in the case of cultured liver cells treated with ethanol. Hence, the study was aimed at determining whether ethanol and Liv.52 could modulate PPARgamma and TNFalpha induction in human hepatoma cells, HepG2. The present study with RT-PCR and confocal microscopy experiments showed that ethanol (100 mM) induced suppression of PPARgamma expression in HepG2 cells. The ethanol-induced PPARgamma suppression was abrogated by Liv.52. Moreover, Liv.52 also induced upregulation of PPARgamma mRNA in liver cells as compared to the untreated cells. Further, 100 mM ethanol has also induced TNFalpha gene expression in HepG2 cells and interestingly Liv.52 abolished ethanol-induced TNFalpha. The study also shows that Liv.52 alone downregulated TNFalpha expression in HepG2 cells. Taken together, these findings suggest that Liv.52 is capable of attenuating ethanol-induced expression of TNFalpha and abrogating ethanol-induced suppression of PPARgamma in liver cells. These results indicate that Liv.52-induced PPARgamma expression and concomitant suppression of ethanol-induced elevation of TNFalpha in HepG2 cells suggest the immunomodulatory and hepatoprotective nature of Liv.52.

Antitumor activity of Hypericum hookerianum against DLA induced tumor in mice and its possible mechanism of action.

A large number of plants belonging to the genus Hypericum and their phytoconstituents are known to possess potent anticancer nature. Earlier studies from our laboratories indicated a strong cytotoxic nature of the methanol extract of Hypericum hookerianum stem (MEHH). In the present study, the in vivo antitumor activity of MEHH against the Dalton's lymphoma ascitic (DLA) model was determined at 100 and 200 mg/kg body weight given orally for 10 days. The results indicate that administration of the extract not only increased the survival of animals with ascites tumor, decreased the body weight induced by the tumor burden and reduced the packed cell volume and viable tissue cell count, but also altered many hematological parameters changed during tumor progression indicating the potent antitumor nature of the extract. Hematological and biochemical analysis were carried out to prove the anticancer and antioxidant nature of the extract.

Antitumor activity and antioxidant status of the methanol extract of Careya arborea bark against Dalton's lymphoma ascites-induced ascitic and solid tumor in mice.

Based on the ethnomedical use of Careya arborea Roxb bark in the treatment of tumors, the present study was carried out to evaluate the anticancer potentials against Dalton's lymphoma ascites (DLA)-induced ascitic and solid tumors. The methanol extract of its bark given orally to mice at the dose of 250 or 500 mg/kg body weight for 10 days caused significant reduction in percent increase in body weight, packed cell volume, and viable tumor cell count when compared to the mice of the DLA control group. Restoration of hematological and biochemical parameters towards normal was also observed. Histological observations of liver and kidney also indicated repair of tissue damage caused by tumor inoculation. The extract at the dose of 5 or 25 mg/kg body weight given i.p. daily for 14 days significantly reduced the solid tumor volume induced by DLA cells.

The cytotoxic activity of the total alkaloids isolated from different parts of Solanum pseudocapsicum.

The total alkaloid fractions of the methanolic extracts of the leaves, ripe fruits, roots, seeds and stem of Solanum pseudocapsicum were subjected to in-vitro cytotoxicity, short-term toxicity and long-term survival studies. All the five fractions exhibited potent activity. The total alkaloid fraction of leaves was found to be the most potent. The HT-29 cell line was the most sensitive to the fractions. The cytotoxic concentration (CTC(50)) values for all these fractions ranged between 0.39-0.91, 0.68-2.8, 0.92-3.56, 4.05-8.2, 3.28-5.65 and 0.95-5.55 microg/ml, respectively for HT-29, RD-228, A-549, HEp-2, B(16)F(10) and Vero cell lines. In short-term toxicity studies, the fractions showed 50% viability at 93-128 microg/ml for DLA cells and 141-189 microg/ml for human lymphocytes. In the long-term survival studies on the cell lines RD-228, HEp-2 and Vero, cells retained their regenerative capacities at concentrations below 8 microg/ml. The total alkaloids of the plant, especially from the leaves merit further investigations to identify the active constituents in animal models.