Green-route synthesis and ab-initio studies of a highly efficient nano photocatalyst:Ce/zinc-oxide nanopetals.

In the present work, Zinc-oxide nanostructures and Ce/Zinc-oxide nanopetals were synthesized by a new environmentally friendly green synthesis method using the Withania coagulans plant. Cerium nitrate Ce(NO3)3 and zinc nitrate Zn(NO3)2 were used as precursors. The prepared nanostructures were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and ultraviolet spectroscopy (UV-vis). Crystal planes (100), (002), (101), (102), (110), (103), (200), (112) and (201) at 2θ 31.75°, 34.35°, 36.2°, 47.55°, 56.6°, 62.75°, 66.3°, 67.9°, and 69.09° respectively confirmed the hexagonal wurtzite crystal structure of Zinc-oxide. Angular shifts for Ce1% doped Zinc-oxide and Ce3% doped Zinc-oxide nanopetal nanostructures were observed in the (100) and (101) planes of the crystal. More specifically, using Scherrer's equation, the crystallite sizes of Zinc-oxide, Ce1% doped Zinc-oxide nanopetals, Ce3% doped Zinc-oxide nanopetals, and Ce5% doped Zinc-oxide nanopetals were 16.48 ± 02 nm, 17.8 ± 2 nm, 18.8 ± 2 nm, and 18.87 ± 2 nm, respectively. The pure Zinc-oxide grain had the appearance of a nanoflower. On the other hand, the nanopetal structure of Ce5% doped Zinc-oxide nanopetals had oval-shaped nanopetal morphology. The absorption peaks were observed at 373, 376.4, 377, and 378 nm for Zinc-oxide, Ce1% doped Zinc-oxide nanopetals, Ce3% doped Zinc-oxide nanopetals, and Ce5% doped Zinc-oxide nanopetals, respectively, which results in a progressive redshift. The gap energies of Zinc-oxide, Ce1% doped Zinc-oxide nanopetals, Ce3% doped Zinc-oxide nanopetals, and Ce5% doped Zinc-oxide nanopetals were 2.796, 2.645, 2.534, and 2.448 eV, respectively. Photodegradation under visible light (>400 nm) indicates the high efficiency of the photocatalyst based on Ce5% doped Zinc-oxide nanopetals. DFT calculations, structural changes, charge analysis, and electronic band structures were carried out to confirm the experiment.

Optimizing the effect of plant protease on different properties of analog cheese containing functional corn leachate.

Cheese is produced in different flavors, textures, and forms by coagulating the milk protein casein. This study investigated the possibility of producing analog cheese by using corn steep liquor with Withania coagulans extract (WCE) and adding Eryngium planum extract (EPE) and Origanum majorana extract (OME) as functional ingredients. Different physicochemical, microbial, texture, and sensory properties of the samples were evaluated. The results obtained for moisture factor, fat, ash, water content, L*, b*, firmness, overall form, Lactobacillus and overall acceptance of the effect of all three process variables, pH, and acidity show that only the effect of WCE and OME is significant. Also, the protein of the samples was significant only on WCE and EPE (p < .001). The results showed that an increase in the levels of independent variables resulted in an increase in the amount of moisture, ash, protein, Lactobacillus, and b* and a decrease in fat, syneresis, texture properties, coliform, and lightness. Evaluation of the overall acceptance showed that consumer acceptance increased with the increase in WCE, but it initially increased and then decreased with the increase in EPE and OME levels. Finally, the samples containing 1.5% WCE, 1% EPE, and 0.5% OME were chosen as the optimized ones.

Withania coagulans fruit extract: a possible useful additive in ameliorating growth and immunity of Labeo rohita (Hamilton, 1822).

This study aimed to assess the Withania coagulans fruit extract's effects on growth and haematological parameters of Labeo rohita. Healthy fish (n = 120) were divided in four groups. Experimental diet was prepared with different extraction of W. coagulans (0%, 1%, 1.5% and 2%). Diet was provided for eight weeks to all groups except the control group which fed on basal diet. The 1.5% and 2% W. coagulans extracts lead improvement in feed conversion ratio and specific fish growth rate; haematological parameters in fish group fed with 2% of W. coagulans showed increased in WBC, RBC, Hb, MCHC, PCV. The values of plasma proteins and lysozyme activity were higher in the experimental group compared control group. Fish fed with W. coagulans extracts showed increased of immunity and less mortality respect to others. It suggested that W. coagulans could be considered valuable support for the improve growth and immunity in Labeo rohita.

DPP-4 inhibition mediated antidiabetic potential of phytoconstituents of an aqueous fruit extract of Withania coagulans (Stocks) Dunal: in-silico, in-vitro and in-vivo assessments.

The DPP-4 inhibition is an interesting target for the development of antidiabetic agents which promotes the longevity of GPL-1(Glucagon-like peptide 1). The current study was intended to assess DPP-4(Dipeptidyl Peptidase-4) inhibition mediated antidiabetic effect of phytocompounds of an aqueous fruit extract of Withania coagulans (Stocks) Dunal by in-vitro, in-silico and in-vivo approaches. The phytoconstituents screening was executed by LCMS (Liquid Chromatography with tandem mass spectrometry). The in-vitro and in-vivo, DPP-4 assays were performed by using available kits. The in-vitro DPP-4 activity was inhibited up to 68.3% by the test extract. Accordingly, in-silico determinations of molecular docking, molecular dynamics and pharmacokinetics were performed between the target enzyme DPP-4 and leading phytocompounds. The molecular dynamics authenticated the molecular docking data by crucial parameters of cytosolic milieu by the potential energy, RSMD (Root Mean Square Deviation), RSMF (Root Mean Square Fluctuation), system density, NVT (Number of particles at fixed volume, ensemble) and NPT (Number of particles at fixed pressure, ensemble). Accordingly, ADMET predictions assessed the druggability profile. Subsequently, the course of the test extract and the sitagliptin (positive control), instigated significant (p ≤ 0.001) ameliorations in HOMA indices and the equal of antioxidants in nicotinamide-streptozotocin induced type 2 diabetic animal model. Compassionately, the histopathology represented increased pancreatic cellular mass which caused in restoration of histoarchitectures. It has been concluded that phytoconstituents in W. coagulans aqueous fruit extract can regulate DPP-4, resulting in improved glucose homeostasis and enhanced endocrinal pancreatic cellular mass.Communicated by Ramaswamy H. Sarma.

Hepatoprotective Role Of N-Hexane Fraction And Crude Methanolic Extract Of Withania Coagulans In Balb C Mice.

Background: Withania coagulans is one of the most important medicinal herbs due to its wide range of biological activities. The aim of this study was to compare the hepatoprotective activity of crude methanolic extract versus n- hexane fraction of fruit of Withania coagulans in CCl4 induced liver toxicity. Methods: This study was done on 36 Balb c mice in Department of Anatomy of Khyber Medical College Peshawar. Results: The mean serum of Group 1(control Group) was 33.41±1.82U/L, for Group 2 (CCl4 treated Group) was 89.01±7.51 U/L, for Group 3(low dose Group) was 49.91±3.48 U/L and for Group 4 (High dose Group) was 50.86±4.87 U/L. There was significant difference in the readings of Group1 and Group 2 which indicated CCL4 induced hepatotoxicity in two groups. There was significant difference in the values of Group 3 and Group 4 (p value .000), showing the hepatotoxicity in these Groups was further enhanced. The mean AST at the end of six weeks for Group1 was 26.80±3.21U/L, for Group 2 was 149.01±13.63U/L, for Group 3 (including both low doses) was 70.81±7.92U/L and for Group 4 (High doses group) was 51.01±11.05U/. Conclusion: Withania coagulans both fractions have hepatoprotective effect against CCL4 induced hepatic toxicity in high and low doses in Balb c mice.

Co-Combination of Pregabalin and Withaniacoagulans-Extract-Loaded Topical Gel Alleviates Allodynia and Hyperalgesia in the Chronic Sciatic Nerve Constriction Injury for Neuropathic Pain in Animal Model.

The current study reports the fabrication of co-combination gel using Pregabalin and Withania coagulans fruit extract to validate its effectiveness for neuropathic pain in chronic constriction injury (CCI) rat models. Three topical gels were prepared using Carbopol 934 through a pseudo-ternary phase diagram incorporating the Pregabalin (2.5%), Withania coagulans extract (2%), and co-combination of both Pregabalin (2.5%) and Withania coagulans extract (2%). Gels were characterized. FTIR showed a successful polymeric network of the gel without any interaction. The drug distribution at the molecular level was confirmed by XRD. The AFM images topographically indicated the rough surface of gels with a size range from 0.25 to 330 nm. DSC showed the disappearance of sharp peaks of the drug and extract, showing successful incorporation into the polymeric network of gels. The in vitro drug release of co-combination gel was 73% over 48 h. The mechanism of drug release by combination gel was Higuchi+ fickian with values of n (0.282) and R2 (0.947). An in vivo study for pain assessment via four methods: (i) heat hyperalgesia, (ii) cold allodynia, (iii) mechano-hyperalgesia, and (iv) dynamic mechano-allodynia, confirmed that topical treatment with co-combination gel reduced the pain significantly as indicated by the p value: R1 (p < 0.001), R2 (p < 0.001), R3 (p < 0.015), and R4 (p < 0.0344). The significance order was R2 (****) > R1 (***) > R3 (**) > R4 (*) > R5 (ns).

Preparation, Characterization of Pregabalin and Withania coagulans Extract-Loaded Topical Gel and Their Comparative Effect on Burn Injury.

The current study depicts the comparative effects of nanogel using Withania coagulans extract, pregabalin alone, and a co-combination gel. The gels prepared were then analyzed for conductivity, viscosity, spread ability, globule size, zeta potential, polydispersity index, and TEM. The globule size of the co-combination gel, determined by zeta sizer, was found to be (329 ± 0.573 nm). FTIR analysis confirms the successful development of gel, without any interaction. Drug distribution at the molecular level was confirmed by XRD. DSC revealed no bigger thermal changes. TEM images revealed spherical molecules with sizes of 200 nm for the co-combination gel. In vivo studies were carried out by infliction of third degree burn wounds on rat skin, and they confirmed that pregabalin and Withania coagulans heals the wound more effectively, with a wound contraction rate of 89.95%, compared to remaining groups. Anti-inflammatory activity (IL-6 and TNF-α), determined by the ELISA technique, shows that the co-combination gel group reduces the maximum inflammation with TNF-α value (132.2 pg/mL), compared to the control (140.22 pg/mL). Similarly, the IL-6 value was found to be (78 pg/mL) for the co-combination gel and (81 pg/mL) in the case of the control. Histopathologically, the co-combination gel heals wounds more quickly, compared to individual gel. These outcomes depict that a co-combination gel using plant extracts and drugs can be successfully used to treat burn injury.

In vitro anticancer and antibacterial potentials of selected medicinal plants and isolation and characterization of a natural compound from Withania coagulans.

In the current study, five different plants, Syzygium Cumini, Fagonia cretica, Acacia modesta, Withania coagulans, and Olea europaea aqueous extracts were prepared and applied against the anticancer and antibacterial activities. It was observed that O. Europaea extract shows the highest anticancer activity with cell viability of 21.5%. All the five plants extract was also used against the inhibition of Bacillus subtilis where O. Europaea extract shows a promising inhibitory activity of 3.2 cm followed by W. coagulans. Furthermore, W. coagulans was subjected to the process of column chromatography as a result a withanolide was isolated. The fast atom bombardment mass spectrometry (FAB-MS) and high resolution fast atom bombardment (HRFAB-MS) [M + 1] indicated molecular weight at m/z 453 and molecular formula C28H37O5. The UV-Vis. spectrum shows absorbance at 210 nm suggesting the presence of conjugated system, and Fourier-transform infrared spectroscopy (FTIR) was recorded to explore the functional groups. Similarly, 1D and 2D NMR spectroscopy techniques such as 1H, 13C NMR, correlation spectroscopy (COSY-45°), heteronuclear single quantum correlation (HSQC), heteronuclear multiple bond correlation (HMBC) and Nuclear Overhauser effect Spectroscopy (NOESY) techniques was carried out to determine the unknown natural product. The collective data of all these techniques established the structure of the unknown compound and recognized as a withanolide.

Anticonvulsant activity of methanolic extract of Withania cogulans in mice.

Mental and neurological diseases including depression, Parkinson's disease, dementia, epilepsy, anxiety disorders and bipolar disorders account for a considerable amount of the world's disease burden. Unfortunately, drugs used in the treatment of neurological diseases are expensive, symptomatic and they produce undesirable side effects. People from different cultures prefer to use medicinal plants for the treatment of various ailments ranging from plain to perplex disorders because they are most affordable, cost effective and easily accessible source of treatment in the primary healthcare system throughout the world. Withania coagulans, an erect grayish under-shrub belongs to family Solanaceae. It is common in Pakistan, East India, Iran and Afghanistan. The objective of this study was to analyze the anti-seizure activity of crude methanolic extract of Withania coagulans fruits (MeWc). For screening of this activity, maximal electroshock seizures model (MES) and chemically-induced seizures models were used. In maximal electroshock seizures test MeWc showed significant dose dependent percent protection against hind-limb tonic extension; significant and dose-dependent increase in latency to myoclonic jerks and tonic clonic convulsions and decrease in seizures duration were observed in PTZ-induced seizures. In strychnine-induced convulsions MeWc significantly increased latency to hind-limb tonic extension and percent protection from death in a dose-dependent manner. Thus, it was inferred from the experiments that extract of Withania coagulans showed anticonvulsant activity.

The effects of supplementation of Withania coagulans and α-tocopherol acetate in diets containing oxidised oil on growth performance, immune response and antioxidant indices in broiler chickens.

This experiment was designed to study the effects of a hydroalcoholic fruit extract from Withania coagulans (WC) and α-tocopherol acetate (α-Toc) in diets containing oxidised oil on growth performance, immune response and antioxidant indices of broiler chickens. The experiment was arranged 2 × 3 × 2 factorial comprising the oxidised oil (0% and 2% diet), WC (0, 100 and 200 mg/kg diet) and α-Toc (0 and 200 mg/kg diet). A total of 600 1-day-old male broiler chickens of the Ross strain were randomly allocated to 12 treatments with 5 replicates of 10 birds each. On day 42, one bird from each replicate was sacrified to measure the lymphoid organ weights, intestinal morphometric characteristics, and malondialdehyde content in the thigh meat. Dietary inclusion of oxidised oil significantly reduced the body weight gain in birds receiving no dietary WC (p < 0.05). In contrast, dietary supplementation of WC at 100 mg/kg diet in birds fed with the diets lacking in WC and oxidised oil resulted in significant increase in body weight gain (p < 0.05). Dietary supplementation of WC significantly increased the total anti-sheep red blood cell titre (p < 0.01). The birds fed with the dietary oxidised oil revealed significantly higher values of malondialdehyde and lower glutathione peroxidase and superoxide dismutase activities (p < 0.05). Feeding diets containing WC resulted in a significant decrease in malondialdehyde content in thigh meat while conversely, increased the glutathione peroxidase and superoxide dismutase activities (p < 0.05). The findings indicated that the dietary inclusion of WC could be recommended as a potent alternative to synthetic compounds in order to improve broilers performance, immunity and meat quality under oxidative stress conditions.

Effect of Fortified Feed with Phyto-Extract on the First Physical Barrier (Mucus) of Labeo rohita.

The aim of the current study was to assess the effect of two different fortified feeds with different concentrations of two important medicinal plants (Withania coagulans and Zingiber officinale) on the mucosal immunity of Labeo rohita. After a dietary intervention, mucus was tested against five pathogenic bacteria (in-vitro), while experimental fish were tested against the ectoparasite (Lernaea) (in-vivo). Our results revealed that all fish groups fed with different concentrations (1, 1.5, and 2%) of Z. officinale had low molecular weight proteins and did not develop any significant signs of parasitic infection, with low mortality rate; whereas the groups that were fed with W. coagulans (particularly with 1% and 2%), including a control group, developed rapid signs of infection with high mortality rate. The highest hemagglutination titer value was recorded for the fish fed with 1% and 1.5% of Z. officinale. The lowest value was found for the fish fed with 2% of W. coagulans. The mucus of all fish of fortified groups was active and inhibited the growth of tested bacterial pathogens as compared to the control group. Further, Z. officinale groups showed greater efficacy against bacteria as compared to the W. coagulans groups. In conclusion, Z. officinale can be considered as a potential and functional ingredient in aquaculture feed. Furthermore, future studies should be conducted to investigate more details on the subject.

Withania coagulans extract attenuates oxidative stress-mediated apoptosis of cerebellar purkinje neurons after ischemia/reperfusion injury.

Cerebral ischemia/reperfusion (I/R) is known to increase reactive oxygen species (ROS) generation, consequences of oxidative stress (OS), and neuronal death in the susceptible brain areas including the cerebellum. Newly, remarkable attention has been paid to a natural diet with the capability to scavenge ROS. Withania coagulans root extract (WCE) is rich in components with antioxidants properties. Therefore, this study aimed to evaluate the effect of WCE on cerebellar Purkinje cells (PCs) against OS-mediated apoptosis after I/R injury. In this experimental study 64 male adult Wistar rats were randomly divided into 4 groups (n = 16) as follows: control, sham, I/R, and WCE 1000 + I/R. I/R animals were pretreated with daily administration of hydro-alcoholic WCE (1000 mg/kg) or distilled water as a vehicle for 30 days before I/R injury. After 72 h, the animals were sacrificed, the cerebellum tissue was removed and used for biochemical (CAT, SOD, GPx, and MDA levels) and histopathological (Nissl and TUNEL staining) assays. Findings showed that the MDA level and the number of apoptotic neurons significantly increased and viable Purkinje neurons decreased in I/R injury (p < 0.05). Administration of 1000 mg/kg WCE reduced MDA level and enhanced antioxidants activity including CAT, SOD, and GPx significantly. In addition, intact surviving PCs increased. At the same time, TUNEL-positive neurons decreased significantly in the WCE pre-treated group (p < 0.05). These findings suggest that WCE can counteract cerebral I/R-induced OS and associated neuronal death by enhancement of ROS scavenging and antioxidant capacity. It appears that pre-treatment with 1000 mg/kg WCE for thirty days can protect PCs against OS-mediated apoptosis after I/R injury.

Evaluation of the Antidiabetic Potential of an Isolated Hydroalcoholic Fraction from the Fruit of Withania coagulans.

The hydro-alcoholic extract of Withania coagulans fruits was investigated for preliminary phytochemical screening and characterized by high-performance thin-layer chromatography. Column chromatography of the hydro-alcoholic extract of W. coagulans eluted with four different combinations of ethyl acetate and methanol yielded four fractions (WCF01, WCF02, WCF03, and WCF04). One of these fractions, WCF02, significantly (P < 0.05) inhibited in vitro α-amylase and α-glucosidase activity with IC50 values of 104.71 µg/mL and 70.79 µg/mL, respectively. WCF02 further reduced blood glucose levels in comparison to control in the starch tolerance test. The extract showed a relative dose-dependent effect. It was observed that none of the extracts could delay the peak blood glucose that was achieved after 60 min of carbohydrate challenge, but these blunted the glycemic peak.

Physiological and anti-oxidative response of biologically and chemically synthesized iron oxide: Zea mays a case study.

The synthesis methodology, particle size and shape, dose optimization, and toxicity studies of nano-fertilizers are vital prior to their field application. This study investigates the comparative response of chemically synthesized and biologically synthesized iron oxide nanorods (NRs) using moringa olefera along with bulk FeCl3 on summer maize (Zea mays). It is found that FeCl3 salt and chemically synthesized iron oxides NRs caused growth retardation and impaired plant physiological and anti-oxidative activities at a concentration higher than 25 mg/L due to toxicity by over accumulation. While iron released form biologically synthesized NRs have shown significantly positive results even at 50 mg/L due to their low toxicity, an improved leaf area (13%), number of leaves per plant (26%), total chlorophyll content (80%) and nitrate content (6%) with biologically synthesized NRs are obtained. Moreover, the plant anti-oxidative activity also increased on treatment with biologically synthesized NRs because of their ability to form a complex with metal ions. These findings suggest that biologically synthesized iron oxides NRs are an efficient iron source and can last for a long time. Thus, proving that nanofertilizer are required to have specific surface chemistry to release the nutrient in an appropriate concentration for better plant growth.

Isolation of Antidiabetic Withanolides from Withania coagulans Dunal and Their In Vitro and In Silico Validation.

Withania coagulans (W. coagulans) is well-known in herbal medicinal systems for its high biological potential. Different parts of the plant are used against insomnia, liver complications, asthma, and biliousness, as well as it is reported to be sedative, emetic, diuretic, antidiabetic antimicrobial, anti-inflammatory, antitumor, hepatoprotective, antihyperglycemic, cardiovascular, immuno-suppressive and central nervous system depressant. Withanolides present in W. coagulans have attracted an immense interest in the scientific field due to their diverse therapeutic applications. The current study deals with chemical and biological evaluation of chloroform, and n-butanol fractions of W. coagulans. The activity-guided fractionation of both extracts via multiple chromatographic steps and structure elucidation of pure isolates using spectroscopies (NMR, mass spectrometry, FTIR and UV-Vis) led to the identification of a new withanolide glycoside, withacogulanoside-B (1) from n-butanol extract and five known withanolides from chloroform extract [withanolid J (2), coagulin E (3), withaperuvin C (4), 27-hydroxywithanolide I (5), and ajugin E (6)]. Among the tested compounds, compound 5 was the most potent α-glucosidase inhibitor with IC50 = 66.7 ± 3.6 µM, followed by compound 4 (IC50: 407 ± 4.5 µM) and compound 2 (IC50: 683 ± 0.94 µM), while no antiglycation activity was observed with the six isolated compounds. Molecular docking was used to predict the binding potential and binding site interactions of these compounds as α-glucosidase inhibitors. Consequently, this study provides basis to discover specific antidiabetic compounds from W. coagulans.

Green synthesis of iron oxide nanorods using Withania coagulans extract improved photocatalytic degradation and antimicrobial activity.

Present work compares the green synthesis of iron oxide nanorodes (NRs) using Withania coagulans and reduction precipitation based chemical method. UV/Vis confirmed the sharp peak of Iron oxide NRs synthesized by biologically and chemically on 294 and 278 nm respectively. XRD and SEM showed highly crystalline nature of NRs with average size 16 ± 2 nm using Withania extract and less crystalline with amorphous Nanostructure of 18 ± 2 nm by chemical method. FTIR analysis revealed the involvement of active bioreducing and stabilizing biomolecules in Withania coagulans extract for synthesis of NRs. Moreover, EDX analysis indicates 34.91% of Iron oxide formation in biological synthesis whereas 25.8% of iron oxide synthesis in chemical method. The degradation of safranin dye in the presence of Withania coagulans based NRs showed 30% more effectively than chemically synthesized Nanorods which were verified by the gradual decrease in the peak intensity at 553 nm and 550 nm respectively under solar irradiation. Furthermore, Withania coagulans based NRs showed effective Antibacterial activity against S.aureus and P. aeuroginosa as compared to NRs by chemical method. Finally, we conclude that green synthesized NRs are more effective and functionally more efficient than chemically prepared NRs. Therefore, our work will help the researchers to boost the synthesis of nanoparticles via biological at commercial level.

Isolation, characterization and DFT studies of epoxy ring containing new withanolides from Withania coagulans Dunal.

A new withanolide named as withacoagulin J (1) along with a known withanolide H (2) from Withania coagulans Dunal are reported in this paper. The isolated compounds were elucidated by using 1D-NMR (1H NMR, 13C NMR) and 2D-NMR including homonuclear (COSY, NOESY) and heteronuclear (HSQC, HMBC); along with Mass spectrometry, UV-Visible and IR spectroscopic techniques. The molecular formula based on Fast-Atom Bombardment Mass Spectrometry [FAB-MS (M + 1)] for 1 and 2 were deduced as C28H37O5 and C28H39O6 with m/z values 453.2624 and 471.6041, respectively. The quantum mechanical studies of both compounds are based on DFT calculations. The DFT studies show band gaps of 4.86 and 4.83 eV for 1 and 2, respectively. The band gaps of 1 and 2 reflect high stability and resistivity towards oxidation-reduction reactions. The energies of HOMO and LUMO for compound 1 are -6.11 and -1.25 eV and for compound 2: -6.47 and -1.64 eV respectively. Theoretical and experimental FTIR data closely match for both the compounds which support the high accuracy of the computational protocol selection. Other parameters such as bond lengths, bond angles and dihedral angles for both compounds are also studied.

Green and cost effective synthesis of silver nanoparticles from endangered medicinal plant Withania coagulans and their potential biomedical properties.

Silver nanoparticles (AgNPs) have great potential for their mechanistic role in biomedical researches. Recently, green biosynthetic approaches have been received much attention in plant science for nanoparticles production. Therefore, in the present study AgNPs have been synthesized utilizing in-vitro grown leaf extract of anti-diabetic medicinal plant Withania coagulans Dunal by the reduction of silver nitrate solution. W. coagulans synthesized silver nanoparticles (WcAgNPs) were characterized by UV-visible spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, X-ray powder diffraction and Fourier transform Infra-Red spectroscopy. All cumulative results showed that WcAgNPs were ~14 nm in size having spherical shape with face centered cubic structure. High performance liquid chromatography confirmed the involvement of withanolides in AgNPs synthesis as a reducing/capping agent. Synthesized WcAgNPs showed greater antioxidative potential when compared with W. coagulans leaf extract. WcAgNPs have efficient antimicrobial potential and suppresses the growth of both gram positive and gram negative bacteria. In our finding we also observed cytotoxicity of WcAgNPs against SiHa (cervical cancerous, hyper-triploid) cell lines and apoptosis in SiHa cells after 48 hour incubation with 13.74 µg ml-1 (IC50) concentration of WcAgNPs. As results suggested, this is the first report which explain that W. coagulans leaf extract have potential as bio-reducing agent for synthesis of silver nanoparticles, which can be exploited as anti-oxidant, antimicrobial and anti-cancerous agent and depicting an effective way for utilizing bioactive resources in restoration of medicinal properties of this plant with high efficacy.

Drier Climatic Conditions Increase Withanolide Content of Withania coagulans Enhancing Its Inhibitory Potential Against Human Prostate Cancer Cells.

Prostate cancer is one of the major causes of cancer-related deaths in men and there is a growing interest in identifying natural compounds for its management. We analyzed bioactive withanolides in Withania coagulans from 11 different sites in Pakistan and evaluated the antiprostate cancer activities of leaf extracts from two sites with the greatest amounts. Total withanolide concentration differed by ~ 17-fold between sites, ranging from 1.01 ± 0.01 mg/g dry weight (mean ± SE) at Jand to 16.83 ± 0.02 mg/g at Mohmand Agency. Different tissues varied in their total withanolide content with roots having the least (0.42 ± 0.07 mg/g dry weight) and leaves the most (2.45 ± 0.45 mg/g). We found strong inverse correlations between site annual precipitation versus withanolide amounts in fruits (r = - 0.84, P = 0.001), leaves (r = - 0.88, P < 0.001), roots (r = - 0.91, P < 0.001), and total (r = - 0.89, P < 0.001), but not stems (r = - 0.20, P = 0.556). Extracts made from Mianwali and Mohmand Agency leaves possessed high anticancer activity in terms of increased induction of apoptosis and decreased cell viability, cell proliferation, invasion, and migration of different prostate cancer cell lines. These results are useful for the selection of withanolide-rich germplasm with potent anticancer properties.

Protective Effect of Withania coagulans Fruit Extract on Cisplatin-induced Nephrotoxicity in Rats.

BACKGROUND: Fruits of Withania coagulans (Solanaceae) reported to possess several bioactive compounds as curative agents for various clinical conditions. Cisplatin is a chemotherapeutic drug to treat sarcomas, carcinomas, lymphomas, cervical cancer, germ cell tumors, etc. The major factor that limits its clinical use is its dose-dependent nephrotoxicity. AIM: To explore the nephroprotective effect of W. coagulans extract and its modulatory effects against cisplatin-induced nephrotoxicity and genotoxicity. MATERIALS AND METHODS: W. coagulans fruit extract was quantitatively standardized with withaferin A using high-performance thin-layer chromatography. The subacute toxicity study was performed according to OECD guidelines in experimental rats. Nephrotoxicity in rats was induced by a single dose of cisplatin (6 mg/kg, intraperitoneal). Nephroprotective role of W. coagulans fruit extract at different doses had been evaluated. It includes quantification of serum kidney toxicity markers, renal tissue oxidative stress biomarkers and pro-inflammatory cytokines level, DNA fragmentation assay, and histopathological examination of renal tissue. RESULTS: Withaferin A was found 3.56 mg/g of W. coagulans fruit extract. It significantly prevented the rise in serum urea and creatinine level and also preserve rat kidneys from oxidative stress and free radical induced DNA damage. Histopathological study showed extract treatment eliminates tubular swelling, cellular necrosis, and protein cast deposition in cisplatin treated kidney tissue. It averted the decline in glutathione content, activities of superoxide dismutase and catalase. These parameters were restored to near normal levels by extract in a dose of 400 mg/kg, per oral. Conclusion: It can be justified that W. coagulans possess dose dependent protective effect against cisplatin induced kidney damages, primarily through its free radical scavenging and anti inflammatory activity. SUMMARY: Authentication and standardization of Withania coagulans fruitsSubacute oral toxicity studyEvaluation of nephroprotective activity against cisplatin-induced nephrotoxicityDNA fragmentation assay and histopathological examination of kidney tissue in experimental rats. Abbreviations Used: WHO: World Health Organization, SOD: Superoxide dismutase, CAT: Catalase, HPTLC: High-performance thin layer chromatography, p.o.: Per.oral, i.p.: Intraperitoneal, TNF-α: Tumor necrosis factor-alpha, IL-1ß: Interleukin 1-beta, IL-6: Interleukin-6.