Metabolite profiling and in-vitro colon cancer protective activity of Cycas revoluta cone extract

Abstract

The methanolic extract of Cycas revoluta cone (MECR) was analyzed by GC-MS and UHPLC for metabolite profiling and was evaluated for anti-colon cancer property by using in vitro assays like Cell Viability Assay, Colony Formation Assay, ROS Determination, Flowcytometry, DAPI staining assay, Tunel assay. GC-MS and HPLC analysis confirmed the presence of different phytochemicals in the extract of Cycas revoluta cone. In-vitro studies showed MECR extract showed significant anti-colon cancer activity by reducing proliferation and inducing apoptosis in colon cancer cell (HCT-8) line, but no such activity was seen in normal colon cell (CCD-18Co) line. The investigation confirms that MECR may be a promising candidate in colon cancer protection.     

In Vivo Evalution of Hypoglycemic Activity of <Emphasis Type="BoldItalic">Aloe</Emphasis> spp. and Identification of Its Mode of Action on GLUT-4 Gene Expression In Vitro

The present study evaluated the hypoglycemic activity of Aloe extract on streptozotocin-induced diabetic mice and focuses its effect on GLUT-4 gene expression under in vitro cell-culture system. Administration of extract at the dosage of 130 mg/kg body weight per day for 4 weeks resulted in significant decrease in blood glucose and total cholesterol in streptozotocin (60 mg/kg body weight) induced diabetic mice. The hypoglycemic effect was compared with metformin. The activities of carbohydrate metabolizing enzymes were brought back to near normal level after the treatment and glucose homeostasis was maintained. Lyophilized aqueous Aloe extract (1 mg/ml) upregulated the GLUT-4 mRNA synthesis in mouse embryonic NIH/3T3 cells.     

Comparing the Impact of NIR,Visible and UV Light on ROS Upregulation via Photoacceptors of Mitochondrial Complexes in Normal,Immune and Cancer Cells

The effect of UV/visible/NIR light (380/450/530/650/808/1064 nm) on ROS generation, mitochondrial activity and viability is experimentally compared in human neuroblastoma cancer cells. The absorption of photons by mitochondrial photoacceptors in Complexes I, III and IV is in detail investigated by sequential blocking with selective pharmaceutical blockers. Complex I absorbs UV/blue light by heme P450, resulting in a very high rate (14 times) of ROS generation leading to cell death. Complex III absorbs green light, by cytochromes b, c1 and c, and possesses less ability for ROS production (seven times), so that only irradiation lower than 10 mW cm⁻² causes an increase in cell viability. Complex IV is well-known as the primary photoacceptor for red/NIR light. Light of 650/808 nm at 10–100 mW cm⁻² generates a physiological ROS level about 20% of a basal concentration, which enhance mitochondrial activity and cell survival, while 1064 nm light does not show any distinguished effects. Further, ROS generation induced by low-intensity red/NIR light is compared in neurons, immune and cancer cells. Red light seems to more rapidly stimulate ROS production, mitochondrial activity and cell survival than 808 nm. At the same time, different cell lines demonstrate slightly various rates of ROS generation, peculiar to their cellular physiology.     

Protective Effect of Amber Extract on Human Dopaminergic Cells against 6-Hydroxydopamine-Induced Neurotoxicity

Parkinson’s disease (PD) is the second most common progressive neurodegenerative disease, after Alzheimer’s disease. In our previous study, we found that amber—a fossilized plant resin—can protect cells from apoptosis by decreasing the generation of reactive oxygen species (ROS). In this study, we focused on the effect of amber on 6-hydroxydopamine-induced cell apoptosis in the human neuroblastoma cell line SHSY5Y (one model for PD). Initially, we determined the protective effect of amber on the PD model. We found that amber extract has a protective effect against 6-hydroxydopamine-induced cell apoptosis. The decrease in ROS, cleaved caspase-3, pERK, and extracellular signal-regulated kinase (ERK) protein levels confirmed that amber extract decreases apoptosis via the ROS-mediated ERK signaling pathway. Furthermore, we determined the effects of amber extract on autophagy. The results showed that amber extract increased the levels of LC3II and Beclin-1, suggesting that amber extract can protect neuronal cells against 6-hydroxydopamine-induced cell apoptosis by promoting autophagy.     

Pathobiological role of advanced glycation endproducts via mitogen-activated protein kinase dependent pathway in the diabetic vasculopathy

Advanced glycation endproducts (AGEs) have been reported to play a role in neointimal formation and increase the rate of in-stent restenosis (ISR) in the diabetic coronary artery disease patients treated with stents, but the potential pathogenic mechanisms of AGEs in vascular smooth muscle cell proliferation remain unclear. We sought to determine the AGEs related pathobiological mechanism of diabetic vasculopathy. Rat aortic smooth muscle cell (RAoSMC) culture was done with different concentrations of AGEs and proliferation was assessed. Immunohistochemistry for receptor of AGEs (RAGE) was performed with human carotid atheroma. Western blotting was performed to assess the activation of MAP kinase system in the cultured RAoSMC. AGEs increased RAoSMC proliferation and were associated with increased phosphorylation of ERK and p38 kinase by time and dose dependent manner. The MAP kinase activity was decreased by RNA interference for RAGE. AGEs stimulation increased reactive oxygen species (ROS) generation in cultured RAoSMC. From this study it is concluded that AGEs played a key role in RAoSMC proliferation via MAP kinase dependent pathways. Activation of vascular smooth muscle cell (VSMC) proliferation by MAP kinase system and increased formation of ROS may be the possible mechanisms of AGEs induced diabetic vasculopathy.     

Proliferative and antioxidant activity of Symphytum officinale root extract

The root of Symphytum officinale L. is commonly used in folk medicine to promote the wound healing, reduce the inflammation and in the treatment of broken bones. The objective of our investigation was to analyse the extract from S. officinale in term of its antioxidant activity and the effect on cell viability and proliferation of human skin fibroblast (HSF). Moreover, the quantification of main phenolics and allantoin was conducted using HPLC–DAD method. Five compounds were found: rosmarinic, p-hydroxybenzoic, caffeic, chlorogenic and p-coumaric acid. DPPH, FRAP and TPC assay showed the high antioxidant activity of the extract. MTT test proved the stimulatory effect on cell metabolism and viability of HSF cells. Moreover, no changes in cytoskeleton structure and cells shape were observed. The obtained results indicate that non-toxic extract from S. officinale root has strong antioxidant potential and a beneficial effect on human skin fibroblasts.     

Anti-inflammatory and antioxidant activities of Rungia congoensis,a traditional vegetable consumed by Yombe people from Kongo Central area (DR. Congo)

Rungia congoensis, a traditional vegetable from Kongo Central area (DR. Congo) was studied for establishing microscopic characters and characterised by chromatographic techniques and their in vitro biochemical activities against ROS production were evaluated in cellular models and on an enzyme, myeloperoxidase (MPO), involved in inflammation. Microscopically leaf can be characterised by non-glandular and glandular trichomes, sinuous anticlinal epidermal cells, diacytic stomata and helical vessels. Methanolic extract displayed high cellular antioxidant activity at the concentrations range of 0.1–10 μg mL^?1 and 1–20 μg mL^?1 using lucigenin on neutrophils and DCFH-DA on HL 60, respectively. This extract also showed, more efficient effects on extracellular and intracellular ROS production and MPO activity. Antioxidant and anti-inflammatory activities of R. congoensis were significantly higher, positively correlated with their phytochemical constituents such as flavonoids, iridoids and phenolic acids; and could justify their use as traditional vegetable and potent local nutraceutical resource.     

Delaying of cataract through intervention of Hemidesmus indicus in STZ induced diabetic rats

Hemidesmus indicus (L.) R. Br. was extensively used as hypoglycaemic agent and significance of this plant on secondary complications of diabetes remained unknown. The present study was to investigate the anti-cataractous activity of H. indicus against streptozotocin (STZ)-induced diabetic cataract in rodent model. Root extracts have been prepared and tested for inhibition of rat lens aldose reductase (AR) activity. In addition, its pharmacological potential has been investigated in STZ-induced diabetic cataract. Methanol extract of H. indicus-inhibited AR activity in vitro decreased the blood glucose levels, inhibited the AR activity and delayed the onset and progression of cataract in a dose-dependent manner in in vivo and the antioxidant markers have been normalised. Our results demonstrate that H. indicus has decrease the osmotic stress by inhibiting the AR activity and prevented the loss of antioxidants and delayed the progression of diabetic cataract in STZ-induced diabetic rats.     

The hypoglycemic and antioxidant activities of garden cress (Lepidium sativum L.) seed on alloxan-induced diabetic male rats

The hypoglycemic and antioxidants activities of the methanol extract of Lepidium sativum seeds was tested in alloxan-induced diabetic male rats. Thirty male albino rats weighing 190–200 g were divided into five groups as follows: negative control, positive control and three diabetic groups treated with three concentrations of L. sativum methanol extract for four weeks. Induction of hyperglycemia in the positive control group increased blood glucose, glycated hemoglobin A1c, immunoglobulins, liver enzyme, lipid peroxide and kidney function, total cholesterol, low-density lipoproteins, very low-density lipoproteins and decreased antioxidants and high-density lipoproteins compared with the negative control. Furthermore, pancreas tissues showed pathological changes compared with the negative control. Treating the diabetic rats with L. sativum methanol extract decreased blood sugar and restored all biochemical and histological changes to the normal. It could be concluded that L. sativum methanol extract succeeded in controlling diabetes, increasing antioxidants and ameliorating lipid profile.     

Eco-friendly synthesis of ZnO nanorods using Cycas pschannae plant extract with excellent photocatalytic,antioxidant, and anticancer nanomedicine for lung cancer treatment

The tunable ZnO nanorods (NRs) are produced due to the phytochemicals present in Cycas pschannae leaves which act as reducing and stabilizing agents. The confirmations of the ZnO NRs were validated using different characterization techniques: X-ray diffraction, Fourier transform infrared spectroscopy, Brunauer, Emmett and Teller (BET), scanning electron microscopy–Energy Dispersive X-Ray Analysis (EDX), UV–visible spectroscopy, Raman spectroscopy, and transmission electron microscopy. The ZnO NRs show unique surface area and low particle size. Photocatalytic activity was measured and found to be 50.75% at low concentrations and 78.33% at high concentrations. The antioxidant activity of the ZnO NRs also showed promising results for their use in free radical scavenging. In vitro toxicity studies using zebrafish embryos was performed to evaluate the toxic nature of it and the obtained result confirmed its non-toxic nature. In addition, ZnO anticancer potential was verified using the A549 lung cancer cell line. Cytotoxic assessments of ZnO NRs were performed via 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutral red uptake assays to examine the cell death cycle on the A549 lung cancer cell. Dose-dependent apoptosis and necrosis were confirmed by Lactate dehydrogenase (LDH) assay. It was also confirmed that ZnO NRs induce Reactive oxygen species (ROS) and apoptosis inside cancer (A549) cells via different intrinsic gene expression. Thus, based on this research it is evident that an effective ecofriendly, nontoxic potential anticancer drug can be synthesized using C. pschannae leaf extract.     

Hypoglycaemic effects of tea extracts and ent-kaurenoic acid from Smallanthus sonchifolius

Hypoglycaemic activity was observed in normoglycaemic mice orally administered with the aqueous Smallanthus sonchifolius leaf tea extract, alloxan-induced diabetic mice orally administered with ent-kaurenoic acid (1), and normoglycaemic mice intraperitoneally administered with 1 from S. sonchifolius leaves. A single dose administration of 50 mg kg(-1) BW yacon leaf tea extract demonstrated immediate but relatively short hypoglycaemic activity, with significant effects observed during 1-2 h. Similarly, administration with 100 mg kg(-1) BW yacon leaf tea extract obtained by heavy stirring in hot water demonstrated a more potent activity compared to the positive control at 1.5-2.0 h. Oral administration of 1 did not affect the blood glucose level of the alloxan-induced diabetic mice, but a single intraperitonial injection of 10 mg kg(-1) BW in normoglycaemic mice had consistent percent blood glucose reduction persisting from 1 to 2 h observation periods.     

Neuroprotective effects of Cassia tora against paraquat-induced neurodegeneration: relevance for Parkinson’s disease

The aim of the present study was to determine whether Cassia tora extracts could reverse the oxidative stress-induced neurodegeneration in a Parkinson’s disease in vitro model. The leaves were treated with ethyl acetate (CtEA) or methanol (CtME). The extracts were first analysed by HPLC for their phenolic content and then tested for their neuroprotective effects in human SK-N-SH neuroblastoma cells. Cells were pre-treated with various concentrations of extracts followed by incubation with paraquat (14 μM). Firstly, pre-treatment of SK-N-SH cells with 100 μg/mL of CtEA or CtME significantly reduced the paraquat-induced production of reactive oxygen species. Furthermore, both CtEA and CtME reduced the paraquat-induced apoptosis. Moreover, there was a significant reduction of paraquat-induced DNA damage in SK-N-SH cells pre-treated with CtEA or CtME. Finally, both extracts significantly inhibited paraquat-dependent lipid peroxidation. Altogether, these in vitro data establish C. tora as a possible anti-Parkinson natural remedy.     

Speciation of organic and inorganic arsenic by HPLC-HG-ICP

This paper deals with the application of high performance liquid chromatography (HPLC), hydride generation (HG) and inductively coupled plasma atomic emission spectrometry (ICP) to determine four species of arsenic: As(III), As(V), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA). The coupling conditions of HPLC-HG-ICP are given. Two anionic exchange columns (Nucleosil-5SB and Hamilton PRP X-100) are tested and the separation conditions optimized. Two acids (H₂SO₄ and HCl at different concentrations) are tested to obtain the hydrides. The proposed method is applied to determine four arsenic species in a synthetic matrix simulating a fish extract.     

Green synthesis of silver nanoparticles by using eugenol and evaluation of antimicrobial potential

The importance of green synthesis was revealed with advantages such as: eliminating the use of expensive chemicals; consume less energy; and generate environmentally benign products. With this aim, silver nanoparticles (AgNPs) were synthesized by using isolated eugenol from clove extract. Its antimicrobial potential was determined on three different microorganisms. Clove was extracted and eugenol was isolated from this extract. Green synthesis was performed and an anti‐microbial study was performed. All extraction and isolation analyses were performed by high‐performance liquid chromatography (HPLC); identification and confirmation were achieved using liquid chromatography–mass spectrometry (LC–MS); and scanning electron microscopy was used for characterization. Both HPLC and LC–MS analyses showed that eugenol obtained purely synthesized AgNPs and 20‐25‐nm‐sized and homogeneous shaped particles seen in images. The antimicrobial effects of AgNPs at eight concentrations were determinated against Staphylococcus aureus, Escherichia coli and Candida albicans, and maximum inhibition zone diameters were found as 2.6 cm, 2.4 cm and 1.5 cm, respectively. The results of the antimicrobial study showed that eugenol as a biological material brought higher antimicrobial effect to AgNPs in comparison to the other materials found in the literature.     

Antidiabetic properties and mechanism of action of Gynura procumbens water extract in streptozotocin-induced diabetic rats

Gynura procumbens (Lour.) Merr (family Compositae) is cultivated in Southeast Asia, especially Indonesia, Malaysia and Thailand, for medicinal purposes. This study evaluated the in vivo hypoglycemic properties of the water extract of G. procumbens following 14 days of treatment and in vitro in RIN-5F cells. Glucose absorption from the intestines and its glucose uptake in abdominal skeletal muscle were assessed. The antidiabetic effect of water extract of G. procumbens leaves was investigated in streptozotocin-induced diabetic rats. The intraperitoneal glucose tolerance test (IPGTT) was performed in diabetic rats treated with G. procumbens water extract for 14 days. In the IPGTT, blood was collected for insulin and blood glucose measurement. After the IPGTT, the pancreases were collected for immunohistochemical study of β-cells of the islets of Langerhans. The possible antidiabetic mechanisms of G. procumbens were assessed through in vitro RIN-5F cell study, intestinal glucose absorption and glucose uptake by muscle. The results showed that G. procumbens significantly decreased blood glucose levels after 14 days of treatment and improved outcome of the IPGTT. However, G. procumbens did not show a significant effect on insulin level either in the in vivo test or the in vitro RIN-5F cell culture study. G. procumbens also showed minimal effects on β-cells of the islets of Langerhans in the pancreas. However, G. procumbens only significantly increased glucose uptake by muscle tissues. From the findings we can conclude that G. procumbens water extract exerted its hypoglycemic effect by promoting glucose uptake by muscles.     

Phytochemical analysis and antidiabetic efficacy of Morus rubra

Morus rubra (red mulberry) is a folk medicine used in management of diabetes. The aim of this article to investigate the antidiabetic activity of hydro alcoholic leaf extracts of Morus rubra in the sprague dawley male rats (SD) model. Authentication of plant material, total Phenolic contents, total flavonoid contents, HPLC analysis, in-vitro antioxidant activity, acute toxicity, in-vivo oral glucose tolerance test, in-vivo STZ induced antidiabetic activity along with biochemical parameters (triglycerides, low-density lipoprotein, high-density lipoprotein, total cholesterol, serum glutamate phosphotransferase, serum glutamate oxaloacetic acid transferase, urea and creatinine). The total phenolic contents 60.32 ​± ​0.80 ​mg/g GAE (gallic acid equivalent) were found higher in ethanolic extract. The total flavonoids 16 ​± ​0.42 ​mg/g QE (quercetin equivalent) were found in petroleum ether extract. M. rubra leaf extracts were tested by HPLC for qualitative and quantitative analysis. 100 ​mg of the dried powder of Morus leaves was dissolve in methanol: water (80:20, v/v) for analysis. Chromatograms of HPLC show the presence of chlorogenic acid 0.55 mg/gm, quercetin 1.90 ​mg/g, isoquercetin 0.68 mg/gm and rutin 0.10 mg/gm. IC₅₀ (Inhibitory concentration) value has been found higher in chloroform extract (92.89 ​μg/ml). Metformin drug is used for positive control, which causes 14.6% improvement in glucose tolerance while M. rubra has been found 11.4. % at the dose of 300 ​mg/kg body wt in normal rats. In STZ treated diabetic rats, it reduces 13.0% blood glucose level while metformin reduces 21.9% up to dosing level of 300 ​mg/kg body wt. It produces reduction in biochemical parameters like, total cholesterol (TC), triglycerides (TG), SGPT, LDL, urea, creatinine and SGOT while HDL has been increased. Results show the hydro-alcoholic leaves extract of Morus rubra exhibited antidiabetic, antioxidant and hypolipidemic properties. This suggests that M. rubra has great potential to develop novel antidiabetic drug to manage this disastrous disease.     

Phytochemical Analysis,Antioxidant, and Wound Healing Activity of Pluchea indica L. (Less) Branch Extract Nanoparticles

Proliferation and migration of keratinocytes and fibroblasts play an important role in cutaneous wound healing, while oral mucosal squamous cell proliferation and migration are crucial for oral wound healing. In this study, the phytochemical profile of Pluchea indica branch ethanolic extract was characterized. The bioactive compound of Pluchea indica branch ethanolic extract was identified and analyzed by the validated HPLC method. The nanoparticles of P. indica branch extract were formulated by solvent displacement method to increase the solubility and the colloidal stability of the extract. The stability of the nanoparticles was investigated by using the dynamic light scattering technique. Effects of P. indica crude extract and nanoparticles on cell viability, proliferation and migration of primary epidermal keratinocytes, human dermal fibroblasts, and oral mucosal keratinocyte cells were investigated by MTT assay and scratch assay, respectively. The results showed that P. indica branch extract contained a high content of total phenolic and total flavonoids. The HPLC analysis revealed that the main compound in the extract was 4,5-O-dicaffeoylquinic acid. The cell viability of the extract and nanoparticles decreased when cells were exposed to a high concentration of extract and nanoparticles. These results demonstrate that P. indica branch extract and extract nanoparticles at specific concentrations possess in vitro wound healing activity and they may be possibly used to treat different types of wounds including dermal and oral mucosal wounds.     

Antihyperglycaemic and hypolipidaemic effects of the methanolic extract of Caralluma tuberculata in streptozotocin-induced diabetic rats

The antihyperglycaemic and hypolipidaemic effects of the methanolic extract of Caralluma tuberculata were investigated in streptozotocin (STZ)-induced diabetic rats. The antihyperglycaemic activity was assessed by the reduction in fasting blood glucose (54% at 4th week) and the peak of blood glucose at 120?min of an oral glucose tolerance test in diabetic rats. Further, the tested extract also increased plasma insulin by 206.8%. The hypolipidaemic action of the extract was evident by the significant decrease in the levels of total cholesterol, triglycerides and LDL-cholesterol by 41.5%, 36.7% and 49.1%, respectively, compared to diabetic rat values. Interestingly, the extract increased the cardio-protective lipid HDL-cholesterol by 147.97% as compared to diabetic rat value. The present data suggests that the methanolic extract of C. tuberculata has both antihyperglycaemic and hypolipidaemic effects in STZ-induced diabetic rats that may need further studies to be used in the management of diabetes and associated hyperlipedaemia.     

Effects of potassium on the ethanol production rate of Saccharomyces cerevisiae carrying the plasmid pCYG4 related with ammonia assimilation

The influence of potassium on ethanol production bySaccharomyces cerevisiae wild type and AR5 cells carrying the plasmid pCYG4 was investigated. This plasmid carries the glutamate dehydrogenase gene conferring an 11-fold higher level of expressed enzyme activity over the wild type cells. All experiments were carried out in batch culture with medium supplemented to different potassium concentrations up to 180 mM. Maximum ethanol production rate was observed in the AR5 cells grown in medium supplemented with 3.5 mM of potassium ions. Glucose uptake rate increased with increasing potassium up to 60 mM, but higher concentrations depressed glucose uptake rate in both strains. Furthermore, the wild type cells showed higher growth rate, ethanol production, and glucose consumption rate than the AR5 cells. These lower rates in the AR5 cells could be explained by repression of potassium uptake by an enhancement of ammonium feeding, and greater energy requirements by these cells due the presence of the plasmid.