Tetrahydropalmatine from medicinal plants activates human glucokinase to regulate glucose homeostasis.

Many synthetic glucokinase activators (GKAs), modulating glucokinase (GK), an important therapeutic target in diabetes have failed to clear clinical trials. In this study, an in silico structural similarity search with differing scaffolds of reference GKAs have been used to identify derivatives from natural product databases. Ten molecules with good binding score and similar interactions to that in the co-crystallized GK as well good activation against recombinant human GK experimentally were identified. Tetrahydropalmatine, an alkaloid present in formulations and drugs from medicinal plants, has not been explored as an antidiabetic agent and no information regarding its mechanism of action or GK activation exists. Tetrahydropalmatine activates GK with EC50 value of 71.7 ± 17.9 µM while lowering the S0.5 (7.1 mM) and increasing Vmax (9.22 µM/min) as compared to control without activator (S0.5 = 10.37 mM; Vmax = 4.8 µM/min). Kinetic data (α and ß values) suggests it to act as mixed, nonessential type activator. Using microscale thermophoresis, Kd values of 3.8 µM suggests a good affinity for GK. In HepG2 cell line, the compound potentiated the uptake of glucose and maintained glucose homeostasis by increasing the expression of GK, glycogen synthase, and insulin receptor genes and lowering the expression of glucokinase regulatory protein (GKRP) and glucagon. Tetrahydropalmatine at low concentrations could elicit a good response by reducing expression of GKRP, increasing expression of GK while also activating it. Thus, it could be used alone or in combination as therapeutic drug as it could effectively modulate GK and alter glucose homeostasis.

Mate calling alters expression of neuropeptide, cocaine- and amphetamine- regulated transcript (CART) in the brain of male frog Microhyla nilphamariensis.

Croaking is a unique component of reproductive behaviour in amphibians which plays a key role in intraspecies communication and mate evaluation. While gonadal hormones are known to induce croaking, central regulation of sound production is less studied. Croaking is a dramatic, transient activity that sets apart an animal from non-croaking individuals. Herein, we aim at examining the profile of the neuropeptide cocaine- and amphetamine-regulated transcript (CART) in actively croaking and non-croaking frog Microhyla nilphamariensis. In anurans, this peptide is widely expressed in the areas inclusive of acoustical nuclei as well as areas relevant to reproduction. CART immunoreactivity was far more in the preoptic area (POA), anteroventral tegmentum (AV), ventral hypothalamus (vHy), pineal (P) and pituitary gland of croaking frog compared to non-croaking animals. On similar lines, tissue fragments collected from the mid region of the brain inclusive of POA, vHy, AV, pineal and pituitary gland of croaking frog showed upregulation of CART mRNA. However, CART immunoreactivity in the neuronal perikarya of raphe (Ra) was completely abolished during croaking activity. The data suggest that CART signaling in the brain may be an important player in mediating croaking behaviour in the frog.

Reproductive phase related variations in the expression of neuropeptide, cocaine- and amphetamine- regulated transcript (CART) in the brain and pituitary gland of adult male Microhyla ornata.

Cocaine- and amphetamine-regulated transcript (CART) peptide is a multifaceted neuropeptide involved in several physiological functions including appetite and reproduction. While studies in mammals, aves and fishes suggest evolutionary conserved role of CART, the information in amphibian is scanty. We have investigated the reproductive phase related variations of CART in the brain of adult male Microhyla ornata. Seasonal changes in the expression of CART peptide were noticed in the brain and pituitary of M. ornata. Significant differences were observed in the nucleus infundibularis ventralis (NIV), epiphysis (E), anteroventral tegmental region (AV), raphe nucleus (Ra) of the brain and pars intermedia (PI), pars distalis (PD) of the pituitary. Compared to the pre-breeding and post-breeding seasons, increase in CART immunoreactivity was seen in E, NIV, AV, Ra of brain and PI, PD of pituitary gland of animals collected during breeding season. Similarly, highest mRNA levels of CART were also observed in the breeding season in the middle region of brain that includes hypothalamus and pituitary gland. Variation in the levels of CART peptide and mRNA in the brain of M. ornata suggests its conserved role in seasonal control of appetite and reproduction.

Cocaine and amphetamine regulated transcript peptide (CART) in the tadpole brain: Response to different energy states.

Cocaine- and amphetamine-regulated transcript peptide (CART) is an anorexigenic neuropeptide known to play a key role in energy homeostasis across the vertebrate phyla. In the current study, we have investigated the response of the CART immunoreactive system to varying energy states in the brain of a tadpole model. The pro-metamorphic tadpoles of Euphlyctis cyanophlyctis were fasted, or intracranially injected with glucose or 2-deoxy-d-glucose (2DG; an antagonist to glucose inducing glucoprivation) and the response of the CART containing system in various neuroanatomical areas was studied using immunohistochemistry. Glucose administration increased the CART immunoreactivity in the entopeduncular neurons (EN), preoptic area (POA), ventral hypothalamus (vHy) and the Edinger Westphal nucleus (EW) while CART positive cells decrease in response to fasting and glucoprivation. A substantial decrease in CART was noted in the EW nucleus of tadpoles injected with 2DG. These regions might contain the glucose-sensing neurons and regulate food intake in anurans. Therefore, we speculate that the function of central CART and its antagonistic action with NPY in food and feeding circuitry of anurans is evolutionary conserved and might be responsible for glucose homeostasis.

Brain profiling of endogenous Neuropeptide Y (NPY) in distinct reproductive phases of adult male Microhyla ornata.

Neuropeptide Y(NPY) is known to play a pivotal role in various physiological functions including appetite and reproduction. While studies in mammals, fishes and reptiles suggest a temporal and evolutionary conserved role of NPY, the information in amphibian is scanty. We have investigated the reproductive phase related variations of NPY in the brain of Microhyla ornata (M. ornata), using immunohistochemistry and reverse transcription quantitative PCR (RT-qPCR). The highest expression of NPY peptide was observed in the preoptic area (Poa), nucleus infundibularis ventralis (NIV) and nucleus reticularis isthmi (NRIS) of M. ornata in breeding season compared to pre-breeding as well as post-breeding season. In parallel, highest mRNA levels of NPY were also observed in the breeding season in the middle region of brain that includes hypothalamus of M. ornata. Variation in the levels of NPY peptide and mRNA levels in the brain of M. ornata point towards seasonal control of appetite and reproduction.

Sex-specific distribution of Neuropeptide Y (NPY) in the brain of the frog, Microhyla ornata.

Neuropeptide Y (NPY) is involved in sex-specific behavioural processes in vertebrates. NPY integrates energy balance and reproduction in mammals. However, the relevance of NPY in reproduction of lower vertebrates is understudied. In the present study, we have investigated neuroanatomical distribution and sex-specific differences of NPY in the brain of Microhyla ornata using immunohistochemistry and quantitative real time PCR. NPY is widely distributed throughout the brain of M. ornata. We observed NPY immunoreactivity in the cells of the nucleus accumbens, striatum pars dorsalis, dorsal pallium, medial pallium, ventral pallium, bed nucleus of stria terminalis, preoptic nucleus, infundibular region, median eminence and pituitary gland of adult M. ornata. A higher number of NPY- immunoreactive cells were observed in the preoptic nucleus (p < .01), nucleus infundibularis ventralis (p < .001) and anteroventral tegmental nucleus (p < .001) of the female as compared to that of the male frog. Real-Time PCR revealed higher mRNA levels of NPY in the female as compared to male frogs in the mid-brain region that largely contains the hypothalamus. Sexual dimorphism of NPY expression in M. ornata suggests that NPY may be involved in the reproductive physiology of anurans.

Response of NPY immunoreactivity in the tadpole brain exposed to energy rich and energy depleted states.

The central control of feeding in animals depends upon the alternating actions of orexigenic and anorectic peptides. Studies at understanding the food intake mechanisms have emphasised the role of Neuropeptide Y as a potent orexigenic peptide in the brain. The aim of this study is to investigate the response of NPY system to positive and negative energy states and elucidate a holistic response of NPY expression throughout the brain of a tadpole model. The pre-metamorphic tadpoles of Euphlyctis cyanophlyctis were subjected to fasting, or intra-cranially injected with glucose or 2-deoxy-d-Glucose (2DG)-a metabolic antagonist of glucose and the response of the NPY system in the entire brain was studied using immunohistochemistry. Glucose injections reduced the basal expression of NPY- immunoreactive perikarya (upto 20%) in the olfactory bulb, nucleus pre-opticus, infundibulum, raphe nucleus and the distal lobe of pituitary. These regions responded to the intracranial injections of 2DG by increasing the expression of NPY up to 30%. Animals deprived of food also possessed the same response except that the increase was much intense in the 2DG injected tadpoles. Our observations lead us to the conclusion that NPY containing neurons in the discrete brain areas may be involved in the maintenance of glucose homeostasis in amphibians and, since these regions also contain the glucose sensing neurons, we further suggest that the release of NPY might be regulated by the glucose sensing neurons of the brain.

NPY up-regulation in the tadpole brain of Euphlyctis cyanophlyctis during osmotic stress.

Most of the amphibians breed in temporary ponds vulnerable to occasional desiccation, thus, leaving their larvae exposed to stressful fluctuations in various environmental parameters including salinity. These animals possess a well suited central adaptive mechanism to adapt to these alterations. Neuropeptide Y (NPY), a 36 amino acid neurotransmitter, has been reported to antagonize various neuropsychological consequences of stress within the mammalian brain. Osmotic regulation of NPY in the hypothalamo-neurohypophysial pathway of mammalian brain is also known. Although the molecule possesses an extensive distribution in the brain of amphibians, its functional association is not well understood. We have investigated the endogenous response of NPY-ergic system to osmotically stressful conditions in the brain of Indian skipper frog-Euphlyctis cyanophlyctis tadpoles. Using Immunohistochemistry, we observed an up-regulation of NPY immunoreactivity (NPY-ir) in the brain of tadpoles exposed to stressful salt concentrations. A significant increase of NPY-ir occurred in the pallium and septum regions of telencephalon; preoptic area, epithalamic, thalamic and hypothalamic parts of diencephalon. Most of the regions are implicated in the modulation of stress and anxiety related brain functions and have also been shown to respond to the salinity stress in mammals. In addition, NPY producing neurons in pre-optic and hypothalamic parts show a close co-existence with the vasopressin-ergic neurons. Thus, our study suggests a possible role of NPY in stabilizing the neuro-endocrinological consequences of osmotic stress in an amphibian brain.

Localization and distribution of superoxide dismutase-1 in the neural tube morphogenesis of chick embryo.

Superoxide dismutase 1 (SOD- 1) is an antioxidant enzyme that regulates the levels of Reactive oxygen species (ROS) by catalyzing the conversion of superoxide radical into hydrogen peroxide (H2O2) and oxygen. ROS are known to play a significant role in various cellular processes, via redox modification of a variety of molecules that participate in signaling pathways involved in this processes. As the levels of ROS in cells are controlled by the levels of antioxidant enzymes, thus SOD-1 may be indirectly involved in regulating different cellular processes by maintaining the required levels of H2O2. Therefore, in the present study we have investigated the possible involvement of SOD- 1 in the neurulation during the development of chick embryo. During gastrulation, SOD- 1 immunoreactivity was observed throughout the ectoderm and cauda mesoderm areas, however, its presence during neurulation was restricted to certain areas of neural tube particularly in the dorsal neural tube where neural tube closure takes place. Assaying enzyme activity revealed a significant increase in the SOD activity during neurulation. Further, inhibition of SOD- 1 by Diethyldithiocarbamate (DDC) induced abnormalities in the development of the neural tube. SOD- 1 inhibition specifically affected the closure of neural tube in the anterior region. Thus, here we report the presence of SOD- 1 mainly in the ectoderm and tissues of ectodermal origin during gastrulation to neurulation which suggests that it may be involved in the regulating the cellular processes during neural tube morphogenesis.

Neuropeptide Y in the brain of Euphlyctis cyanophlyctis tadpoles responds to hypoxic stress.

Neuropeptide Y (NPY) has emerged as a novel peptide to antagonize various physiological consequences of stress within a mammalian brain. Hypoxia induced neuropeptide Y release in mammalian systems is well established. However, the possible role of NPY in regulating the effects of oxygen variation in lower vertebrates has not been investigated. We have studied the distribution and neuro-anatomical expression of NPY in the brain of Euphlyctis cyanophlyctis tadpoles, exposed to normal and reduced oxygen levels using immunohistochemistry. Animals exposed to hypoxia (<2mg/ml) exhibited a significant amplification of NPY-immunoreactivity throughout the brain. Increased NPY-ir perikarya appeared in all the sub-divisions of pallium, septum and preoptic area of telencephalon; suprachiasmatic nucleus, central and lateral thalamus, infundibulum and habenular regions of diencephalon; and nucleus cerebella and medulla of rhombencephalon. Most of these regions form the stress and anxiety regulating centers of a vertebrate brain and some of the parallel regions also respond to respiratory reflexes in mammals. Hence, our results suggest NPY induced modulation of hypoxia in Euphlyctis cyanophlyctis tadpoles.

The zebrafish goosepimples/myosin Vb mutant exhibits cellular attributes of human microvillus inclusion disease.

Microvillus inclusion disease (MVID) is a life-threatening enteropathy characterised by malabsorption and incapacitating fluid loss due to chronic diarrhoea. Histological analysis has revealed that enterocytes in MVID patients exhibit reduction of microvilli, presence of microvillus inclusion bodies and intestinal villus atrophy, whereas genetic linkage analysis has identified mutations in myosin Vb gene as the main cause of MVID. In order to understand the cellular basis of MVID and the associated formation of inclusion bodies, an animal model that develops ex utero and is tractable genetically as well as by microscopy would be highly useful. Here we report that the intestine of the zebrafish goosepimples (gsp)/myosin Vb (myoVb) mutant shows severe reduction in intestinal folds - structures similar to mammalian villi. The loss of folds is further correlated with changes in the shape of enterocytes. In striking similarity with MVID patients, zebrafish gsp/myoVb mutant larvae exhibit microvillus atrophy, microvillus inclusions and accumulation of secretory material in enterocytes. We propose that the zebrafish gsp/myoVb mutant is a valuable model to study the pathophysiology of MVID. Furthermore, owing to the advantages of zebrafish in screening libraries of small molecules, the gsp mutant will be an ideal tool to identify compounds having therapeutic value against MVID.

Biomarker responses in the earthworm, Dichogaster curgensis exposed to fly ash polluted soils.

Earthworms are globally accepted as a model organism in terrestrial ecotoxicology for assessment of environmental pollution. This study evaluated and compared effects of fly ash polluted soils collected from two geographically different thermal power plants on biomarker responses in the earthworm, Dichogaster curgensis. To evaluate relationship between distance sampling and biomarker responses in the earthworm D. curgensis, soil samples at 0.5, 1 and 3km from thermal plant were analyzed for physico-chemical properties and metal concentrations. Biochemical alterations, lysosomal membrane stability, genotoxic effects, and histological changes were examined on 1, 7, and 14 d of exposure to fly ash contaminated soils collected from different thermal power plants. The activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels were significantly increased, while glutathione reductase (GR) activity was found to be decreased in treated animals. Catalase (CAT) and glutathione-S- transferase (GST) activities were found to be increased initially up to 7d exposure and further decreased on 14d exposure. D. curgensis exposed to fly ash contaminated soils showed significant lysosomal membrane destabilization and DNA damage. Extensive histopathological changes were observed in the tissues of the body wall and intestinal tract of the exposed D. curgensis along with accumulation of heavy metals. These results demonstrate that soil pollution around thermal power plants has adverse biological effects of on the indicator organism D. curgensis and no correlation was found between distance and extent of biological biochemical responses.

Temperature dependent changes in cocaine- and amphetamine regulated transcript (CART) peptide in the brain of tadpole, Sylvirana temporalis.

Cocaine- and amphetamine-regulated transcript peptide (CARTp) has emerged as a novel neurotransmitter in the brain. Although the physiological role of the peptide has been intensely investigated in mammals, its role in amphibians has not been investigated. In the present study, an attempt has been undertaken to study the expression of CART in the tadpole brain of frog Sylvirana temporalis, subjected to thermal stress. Cells with strong CART-immunoreactivity were observed in the nucleus preoptic area (NPO) of tadpoles exposed to high temperature (37±2°C) as compared to those in the tadpoles exposed to low (12±2°C) and normal (24±2°C) temperatures. In the ventromedial thalamic nucleus (VM) and nucleus posterocentralis thalami (NPC), moderate CART-ir cells were observed in the control groups while number of cells and intensity of immunoreactivity was increased in tadpoles at low and high temperatures. In the nucleus infundibularis ventralis (NIV) and raphe nucleus (RA), CART immunoreactivity increased in the low as well as high temperature treated groups. Intensely stained CART cells were observed in the pituitary of tadpoles exposed to high temperature as compared to low temperature and control groups. We suggest that CART system in the brain and pituitary of tadpole may play a very important role in mediating responses to temperature variations in the environment.

Nesfatin-1/nucleobindin-2 like immunoreactivity in the olfactory system, brain and pituitary of frog, Microhyla ornata.

Nesfatin-1 is a recently discovered anorectic protein derived from the precursor nucleobindin-2 (NUCB2). While nesfatin-1 has been widely studied in mammals, and goldfish, there are no reports of nesfatin-1 in amphibians. Using immunohistochemistry and Western blot analysis, this study assessed the distribution of NUCB2/nesfatin-1 in the brain of frog Microhyla ornata. NUCB2/nesfatin-1 like immunoreactivity was found in the olfactory receptor neurons, Bowman's glands and in the olfactory epithelium of medial diverticulum. In the brain, immunoreactive perikarya were seen in the anterior preoptic area, magnocellular nucleus, suprachiasmatic nucleus, ventromedial thalamic nucleus, posterior thalamic nucleus, nucleus infundibularis ventralis and dorsalis, and the cerebellar nucleus. NUCB2/nesfatin-1like immunoreactivity was also detected in the pineal and pituitary glands of frog. The presence of NUCB2/nesfatin-1 in the key brain regions suggest possible roles for this protein in the regulation of physiological processes in frogs.

Ontogeny of cocaine- and amphetamine-regulated transcript peptide in brain of frog, Microhyla ornata.

The cocaine- and amphetamine-regulated transcript (CART) peptide is widely distributed in the brains of adult vertebrates including amphibians. Several physiological roles of CART have been intensely investigated in mammals. Despite these studies, the expression of CART during development of brain has not been studied in amphibians. In the present study, distribution of CART was investigated during development in the post hatched stage 23 to premetamorphic stage 30 of frog Microhyla ornata. CART is expressed as early as in stage 23 in ventral thalamus and rhombencephalon. As development progressed, CART immunoreactivity was observed in the olfactory bulb, telencephalon, rhombencephalon and spinal cord in stage 24. At stage 25, the CART immunoreactivity was observed in the ventromedial thalamic nucleus, posterocentral thalamic nucleus, torus nucleus, central gray and inferior reticular nucleus. In stage 26, CART reactivity was seen in the medial septum, preoptic area, nucleus entopeduncularis, magnocellular nucleus, median eminence, optic tectum, hypophysis and cerebellum. Additionally, CART immunoreactivity was observed in the medial pallium, anterior commissure, nucleus infundibularis dorsalis, ventralis and raphe nucleus at stage 30. The occurrences of CART immunoreactivity at early stage of development suggest that the peptide may have a functional significance during development. The wider appearance of CART in the brain of tadpoles, M. ornata suggests that the peptide may act as a neurohormone during the ontogeny.

Discovering Bisdemethoxycurcumin from Curcuma longa rhizome as a potent small molecule inhibitor of human pancreatic α-amylase, a target for type-2 diabetes.

Curcuma longa rhizome is used extensively in culinary preparations in Far East and South-East Asia. Health benefits of curcuminoids from C. longa as antioxidants, anti-cancer and anti-inflammatory molecules have been well documented. We report here for the first time that Bisdemethoxycurcumin (BDMC) from C. longa, acts as an inhibitor to inactivate human pancreatic α-amylase, a therapeutic target for oral hypoglycemic agents in type-2 diabetes. Bioactivity guided isolation of rhizome isopropanol extract led to the identification by HPLC and NMR of BDMC as a lead small molecule inhibitor of porcine and human pancreatic α-amylase with an IC(50) value of 0.026 and 0.025 mM, respectively. Kinetic analysis revealed that using starch as the substrate, HPA exhibited an uncompetitive mode of inhibition with an apparent K(i) of 3.0 µM. The study gains importance as BDMC could be a good drug candidate in development of new inhibitors of HPA and of functional foods for controlling starch digestion in order to reduce post-prandial hyperglycemia.

Immunohistochemical localization and biochemical changes in catalase and superoxide dismutase during metamorphosis in the olfactory system of frog Microhyla ornata.

Amphibian metamorphosis is characterized by rapid tissue remodeling and drastic changes in the body structure and function. Like other organs, olfactory system also undergoes a dramatic rearrangement as the animal experiences transition from aquatic to terrestrial habitat. Reactive oxygen species (ROS) are known to play an important role during anuran metamorphosis and role of antioxidant enzymes like catalase and superoxide dismutase (SOD) are believed to play a major role in these processes. Therefore, we hypothesize that antioxidant enzymes in the olfactory system may undergo changes that reflect metamorphic processes. Immunohistochemical study revealed the presence of catalase and SOD in the olfactory receptor neurons and also granular reaction in olfactory epithelium of medial diverticulum during metamorphosis. Catalase and SOD immunoreactivity were seen in the epithelium of lateral diverticulum, vomeronasal organ as metamorphosis proceeds and in the apical lining of olfactory epithelium of adult frog. Biochemical study showed that catalase activity gradually increases in the olfactory system from metamorphic stage 40-46 and adult, while SOD activity decreases from stage 40 to 46 and increases in adult. Thus, the localization and relative levels of catalase and SOD during metamorphosis in the olfactory system suggests that these enzymes may be involved in protection from oxidative damage.

Potent α-amylase inhibitory activity of Indian Ayurvedic medicinal plants.

BACKGROUND: Indian medicinal plants used in the Ayurvedic traditional system to treat diabetes are a valuable source of novel anti-diabetic agents. Pancreatic α-amylase inhibitors offer an effective strategy to lower the levels of post-prandial hyperglycemia via control of starch breakdown. In this study, seventeen Indian medicinal plants with known hypoglycemic properties were subjected to sequential solvent extraction and tested for α-amylase inhibition, in order to assess and evaluate their inhibitory potential on PPA (porcine pancreatic α-amylase). Preliminary phytochemical analysis of the lead extracts was performed in order to determine the probable constituents. METHODS: Analysis of the 126 extracts, obtained from 17 plants (Aloe vera (L.) Burm.f., Adansonia digitata L., Allium sativum L., Casia fistula L., Catharanthus roseus (L.) G. Don., Cinnamomum verum Persl., Coccinia grandis (L.) Voigt., Linum usitatisumum L., Mangifera indica L., Morus alba L., Nerium oleander L., Ocimum tenuiflorum L., Piper nigrum L., Terminalia chebula Retz., Tinospora cordifolia (Willd.) Miers., Trigonella foenum-graceum L., Zingiber officinale Rosc.) for PPA inhibition was initially performed qualitatively by starch-iodine colour assay. The lead extracts were further quantified with respect to PPA inhibition using the chromogenic DNSA (3, 5-dinitrosalicylic acid) method. Phytochemical constituents of the extracts exhibiting≥ 50% inhibition were analysed qualitatively as well as by GC-MS (Gas chromatography-Mass spectrometry). RESULTS: Of the 126 extracts obtained from 17 plants, 17 extracts exhibited PPA inhibitory potential to varying degrees (10%-60.5%) while 4 extracts showed low inhibition (< 10%). However, strong porcine pancreatic amylase inhibitory activity (> 50%) was obtained with 3 isopropanol extracts. All these 3 extracts exhibited concentration dependent inhibition with IC50 values, viz., seeds of Linum usitatisumum (540 µgml-1), leaves of Morus alba (1440 µgml-1) and Ocimum tenuiflorum (8.9 µgml-1). Acarbose as the standard inhibitor exhibited an IC50 (half maximal inhibitory concentration)value of 10.2 µgml-1. Phytochemical analysis revealed the presence of alkaloids, tannins, cardiac glycosides, flavonoids, saponins and steroids with the major phytoconstituents being identified by GC-MS. CONCLUSIONS: This study endorses the use of these plants for further studies to determine their potential for type 2 diabetes management. Results suggests that extracts of Linum usitatisumum, Morus alba and Ocimum tenuiflorum act effectively as PPA inhibitors leading to a reduction in starch hydrolysis and hence eventually to lowered glucose levels.

Evaluation of traditional Indian antidiabetic medicinal plants for human pancreatic amylase inhibitory effect in vitro.

Pancreatic α-amylase inhibitors offer an effective strategy to lower the levels of post prandial hyperglycemia via control of starch breakdown. Eleven Ayurvedic Indian medicinal plants with known hypoglycemic properties were subjected to sequential solvent extraction and tested for α-amylase inhibition, in order to assess and evaluate their inhibitory potential on pancreatic α-amylase. Analysis of 91 extracts, showed that 10 exhibited strong Human Pancreatic Amylase (HPA) inhibitory potential. Of these, 6 extracts showed concentration dependent inhibition with IC(50) values, namely, cold and hot water extracts from Ficus bengalensis bark (4.4 and 125 µgmL(-1)), Syzygium cumini seeds (42.1 and 4.1 µgmL(-1)), isopropanol extracts of Cinnamomum verum leaves (1.0 µgmL(-1)) and Curcuma longa rhizome (0.16 µgmL(-1)). The other 4 extracts exhibited concentration independent inhibition, namely, methanol extract of Bixa orellana leaves (49 µgmL(-1)), isopropanol extract from Murraya koenigii leaves (127 µgmL(-1)), acetone extracts from C. longa rhizome (7.4 µgmL(-1)) and Tribulus terrestris seeds (511 µgmL(-1)). Thus, the probable mechanism of action of the above fractions is due to their inhibitory action on HPA, thereby reducing the rate of starch hydrolysis leading to lowered glucose levels. Phytochemical analysis revealed the presence of alkaloids, proteins, tannins, cardiac glycosides, flavonoids, saponins and steroids as probable inhibitory compounds.

Antidiabetic Indian plants: a good source of potent amylase inhibitors.

Diabetes is known as a multifactorial disease. The treatment of diabetes (Type II) is complicated due to the inherent patho-physiological factors related to this disease. One of the complications of diabetes is post-prandial hyperglycemia (PPHG). Glucosidase inhibitors, particularly α-amylase inhibitors are a class of compounds that helps in managing PPHG. Six ethno-botanically known plants having antidiabetic property namely, Azadirachta indica Adr. Juss.; Murraya koenigii (L.) Sprengel; Ocimum tenuflorum (L.) (syn: Sanctum); Syzygium cumini (L.) Skeels (syn: Eugenia jambolana); Linum usitatissimum (L.) and Bougainvillea spectabilis were tested for their ability to inhibit glucosidase activity. The chloroform, methanol and aqueous extracts were prepared sequentially from either leaves or seeds of these plants. It was observed that the chloroform extract of O. tenuflorum; B. spectabilis; M. koenigii and S. cumini have significant α-amylase inhibitory property. Plants extracts were further tested against murine pancreatic, liver and small intestinal crude enzyme preparations for glucosidase inhibitory activity. The three extracts of O. tenuflorum and chloroform extract of M. koenigi showed good inhibition of murine pancreatic and intestinal glucosidases as compared with acarbose, a known glucosidase inhibitor.