Investigation of phytotherapeutic potential of herbal mixtures and their effects on salbutamol induced cardiotoxicity and hyperlipidemia in rabbits.

BACKGROUND: Cardiovascular diseases (CVDs) are the major cause of deaths all over the world. The high level of blood cholesterol and oxidative stress are major risk factors for heart diseases. The phytotherapeutics have attracted attention as potential agents for preventing and treating oxidative stress associated diseases. The objective of present study was to evaluate the synergetic cardio-protective and antilipidemic potential of medicinal plants viz. Coriandrum sativum, Piper nigrum and Cactus grandiflorus. Cardio-protective and anti-lipidemic potential of herbal mixture was evaluated against salbutamol induced cardiotoxicity in rabbits. For this purpose, rabbits were divided into six groups as normal control, salbutamol control, curative and standard drug curative. RESULTS: Salbutamol significantly (p < 0.05) increased the level of serum cardiac biomarkers (ALT, CK-MB, AST and LDH) and lipids (LDL, triglycerides, cholesterol) in rabbits. The prior and post administration of herbal mixture significantly (p < 0.05) lowered the elevated level of serum cardiac biomarkers and lipids equal to normal control. Gross pathological examination revealed that heart of salbutamol control animals became hardened, congested and were enlarged than preventive and curative groups. The phytotherapeutic analysis of medicinal plants revealed the presence of phenols, tannins, alkaloids and steroids. CONCLUSION: The results showed that this herbal mixture has strong cardio-protective and anti-lipidemic potential.

Green synthesized silver nanoparticles: Optimization, characterization, antimicrobial activity, and cytotoxicity study by hemolysis assay.

Green nanotechnology has emerged as a viable option for the production of nanoparticles. The purpose of the current investigation was to synthesize silver nanoparticles (AgNPs) using Eucalyptus camaldulensis and Terminalia arjuna extracts, as well as their combinations, as green reducing and capping agents. The parameters (concentration of silver nitrate solution and plant extract, time, pH, and temperature) were optimized for maximal yields, regulated size, and stability of silver nanoparticles. The ultraviolet-visible spectrophotometer (UV-Vis) and the surface plasmon resonance band (SPR) were used to validate the synthesis of AgNPs. The size, shape, and stability of nanoparticles were assessed using a zeta analyzer and a scanning electron microscope (SEM). The biomolecules responsible for the reduction of silver ion (Ag+) and the stability of silver nanoparticles generated with the plant extracts were identified using Fourier-transform infrared spectroscopy (FTIR). The agar-well diffusion method was used to test the antimicrobial activity of biosynthesized nanoparticles against Bacillus subtilis, Staphylococcus aureus, Pasteurella multocida, and Escherichia coli. When 1 mM of silver nitrate (AgNO3) was added to plant extracts and incubated for 60 min at 75°C in a neutral medium, maximum nanoparticles were produced. Biosynthesized silver nanoparticles were stable, spherical, and monodispersed according to zeta potential and scanning electron microscopy. Silver nanoparticles synthesized with combination 2 and T. arjuna showed the highest zone of inhibition (16 mm) against B. subtilis while combination 3 showed the largest zone of inhibition against S. aureus (17 ± 0.8). It was concluded that greenly produced silver nanoparticles showed good antibacterial activity while causing negligible cytotoxicity.

Biologically Synthesized Peptides Show Remarkable Inhibition Activity against Angiotensin-Converting Enzyme: A Promising Approach for Peptide Development against Autoimmune Diseases.

Angiotensin-converting enzyme (ACE) regulates several biological functions besides its vital role in immune functions. ACE is elevated in immune cells in inflammatory diseases including atherosclerosis, granuloma, chronic kidney disease, and also autoimmune diseases, like multiple sclerosis, rheumatoid arthritis, and type I diabetes. No significant information prevails in the literature regarding the isolation, identification, and profiling of potential ACE inhibitory peptides. In the present study, indigenous crop varieties like seeds (peanut, corn, oat, sunflower, chickpea, parsley, cottonseed, papaya, sesame, and flaxseed) were used to evaluate their ACE inhibition activity. Variables including hydrolysis time, enzyme-to-substrate ratio (E/S), pH, and temperature were standardized to acquire the most suitable and optimum ACE inhibition activity. Seeds of cotton, chickpea, and peanuts displayed remarkably maximum ACE inhibition activity than other plants. The study disclosed that maximum ACE inhibitory activity (86%) was evaluated from cottonseed at pH 8.0, temperature of 45°C, hydrolysis time of 2 hrs, and enzyme to the substrate (E/S) ratio of 1 : 5 followed by peanuts (76%) and chickpea (55%). SDS-PAGE confirmed that vicilin protein is present in cottonseed and peanut seed while cruciferin and napin proteins are present in chickpeas. LC-MS/MS analysis disclosed potential novel peptides in hydrolyzed cottonseed that can be ascribed as potential ACE inhibitors which have never been reported and studied earlier. The current study further showed that cottonseed peptides due to their promising ACE inhibitory activity can be a valuable source in the field of ACE inhibitor development.

Evaluation and optimization of nanosuspensions of Chrysanthemum coronarium and Azadirachta indica using Response Surface Methodology for pest management.

The rapidly emerging field of nanotechnology is considered an important achievement in the agriculture sector to increase the pest mortality rate and improve the crop production. The present study evaluates the novel pesticidal and anti-microbial activities of Chrysanthemum coronarium and Azadirachta indica in the nano-suspensions form. The anti-solvent precipitation method was used to formulate nano-suspensions proposed by Response Surface Methodology (RSM). Physicochemical nature of plant extracts and nano-suspensions was characterized through analysis of Zeta-sizer, FT-IR, and HPLC. Characterization results revealed a minimum particle size of 121.1 and 170.1 nm for Chrysanthemum coronarium and Azadirachta indica, respectively. The pesticidal activity of nano-suspension was performed against red flour beetle (RFB) and lesser grain borer (LGB) pests, which showed the maximum mortality rate of 100% with 100% concentration of plant extracts and nano-suspensions of Chrysanthemum coronarium and Azadirachta indica against both insects. In comparison, the combination of these both plant extracts revealed the maximum 100% mortality with a 50% concentration of nano-suspensions (mixing ratio 1:1) after 72 h. The antibacterial activity showed the maximum zone inhibition of 9.96 ± 0.17 and 14.17 ± 0.50 mm against S.aureus and E. coli with nano-suspension of Chrysanthemum coronarium, and 12.09 ± 0.11 and 14.10 ± 0.49 mm with nano-suspension of Azadirachta indica, respectively. It is concluded that individual nano-suspensions showed better pesticidal as well as antimicrobial activities than combinations. However, the constructed nanosuspension can be applied to control the plant pests and diseases simultaneously.

Isolation and characterization of antihypertensive peptides from soy bean protein

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.

Synthesis and biological evaluation of Ellettaria cardamomum (Cardamom) Phytosomes.

Ellettaria cardamomum (Cardamom) is an excellent antioxidant and its phytochemicals possess astounding detox properties. The bioactives of E. cardamomum have low bioavailability due to complex molecular structure. The main aim of this research was to formulate the E. cardamomum loaded phytosomes to improve the bioavailability with better ACE inhibition potential. The phytosomes were prepared by thin layer hydration technique. Various synthetic parameters for formulation of phytosomes were optimized by response surface methodology. Formulated phytosomes were characterized through spectroscopically and their biological activities eg. ACE inhibition were also investigated. Resultantly, the optimal production of phytosomes was achieved with the equal quantity (300 mg) of E. cardamomum and phospholipids at the time of 20 minutes of hydration. UV-Visible spectra confirmed the chemical and physical interaction between phospholipid and bioactives functional groups. The optimized E. cardamomum phytosome had 71% of entrapment efficiency. The average vesicle size of phytosomes was 577.8 nm with polydispersity index 0.443. SEM analysis revealed that phytosomes were spherical in shape. The phytosomes of E. cardamomum showed higher antioxidant and antimicrobial activities than its crude extract. The ACE inhibition activity of phytosomes (46%) was enhanced than the crude extract (39 %).

The Role of Polyphenol (Flavonoids) Compounds in the Treatment of Cancer Cells.

Cancer remains a second leading cause of deaths and major public health problem. It occurs due to extensive DNA damage caused by ultraviolet radiations, ionizing radiations, environmental agents, therapeutic agents, etc. Among all cancers, the most frequently diagnosed cancers are lung (12.7%), breast (10.9%), colorectal (9.7%), and gastric cancer (7.81%). Natural compounds are most favorable against cancer on the count of their anti-cancerous ability, easy to avail and efficient. Among natural compounds, polyphenols (flavonoids, catechin, hesperetin, flavones, quercetin, phenolic acids, ellagic acid, lignans, stilbenes, etc.) represent a large and diverse group used in the prevention and treatment of cancer. Natural flavonoids are derived from different plant sources and from various medicinal plants including Petroselinum crispum, Apium graveolens, Flemingia vestita, Phyllanthus emblica, etc. Natural flavonoids possess antioxidant, anti-inflammation, as well as anti-cancerous activities through multiple pathways, they induce apoptosis in breast, colorectal, and prostate cancers, lower the nucleoside diphosphate kinase-B activity in lung, bladder and colon cancers, inhibit cell-proliferation and cell cycle arrest by suppressing the NF-kB pathway in various cancers, etc. The current review summarized the anticancer activities of natural polyphenols and their mechanisms of action.

Increased Oral Bioavailability of Piperine from an Optimized Piper nigrum Nanosuspension.

The aim of the present study was to enhance the pharmaceutical potential and oral bioavailability of piperine, which is the bioactive constituent of Piper nigrum, using the nanosuspension approach. Nanoprecipitation, which is a simple and reproducible process, was used for nanosuspension formulation. To prepare a pharmaceutical-grade nanosuspension with the required particle size, important formulation parameters (amount of plant extract, concentration of stabilizer, and antisolvent-to-solvent ratio) were optimized using the central composite design of response surface methodology. The optimized nanosuspension was characterized using scanning electron microscopy, atomic force microscopy, Fourier transform infrared spectroscopy, and in vitro dissolution testing as well as by measuring the zeta potential. In vivo pharmacokinetic studies were conducted to determine the bioavailability of the prepared nanosuspension. Results of the optimization study indicated that 0.13% plant extract, 0.25% stabilizer, and an antisolvent-to-solvent ratio of 10.0 were the best parameters to obtain a homogeneous nanosuspension with the required particle size. The optimized nanosuspension demonstrated a mean particle size, polydispersity index, and zeta potential of 172.5 nm, 0.241, and - 16.6 mV, respectively. The results of the characterization studies illustrated that the nanosuspension was in the nanometer size range and had good surface morphology. The optimized nanosuspension showed a better dissolution rate and a 3.65-fold higher oral bioavailability for the P. nigrum nanosuspension than its coarse suspension. The present outcomes clearly demonstrated that to obtain an effective therapeutic potential, nanoformulation of medicinal plants is a better alternative than conventional dosage forms.

Cardioprotective and Metabolomic Profiling of Selected Medicinal Plants against Oxidative Stress.

In this research work, the antioxidant and metabolomic profiling of seven selected medicinally important herbs including Rauvolfia serpentina, Terminalia arjuna, Coriandrum sativum, Elettaria cardamom, Piper nigrum, Allium sativum, and Crataegus oxyacantha was performed. The in vivo cardioprotective potential of these medicinal plants was evaluated against surgically induced oxidative stress through left anterior descending coronary artery ligation (LADCA) in dogs. The antioxidant profiling of these plants was done through DPPH and DNA protection assay. The C. oxyacantha and T. arjuna showed maximum antioxidant potential, while the E. cardamom showed poor antioxidative strength even at its high concentration. Different concentrations of extracts of the said plants exhibited the protection of plasmid DNA against H2O2 damage as compared to the plasmid DNA merely treated with H2O2. The metabolomic profiling through LC-MS analysis of these antioxidants revealed the presence of active secondary metabolites responsible for their antioxidant potential. During in vivo analysis, blood samples of all treatment groups were drawn at different time intervals to analyze the cardiac and hemodynamic parameters. The results depicted that the group pretreated with HC4 significantly sustained the level of CK-MB, SGOT, and LDH as well as hemodynamic parameters near to normal. The histopathological examination also confirmed the cardioprotective potential of HC4. Thus, the HC4 being safe and inexpensive cardioprotective herbal combination could be considered as an alternate of synthetic drugs.

Synergistic interactions of polyphenols and their effect on antiradical potential.

The aim of the current study was to evaluate interactions among polyphenols from different plants and their effect on antioxidant potential. Different mixtures of plant extracts of Crataegus oxyacantha (C), Elettaria cardamomum (Cr), Terminalia arjuna (T) and Rauvolfia serpentina (R) were prepared and evaluated for total phenolics, flavonoid contents, and antioxidant activity. A correlation was also established between total phenolics, flavonoids and antioxidant activity. Comparative evaluation revealed that phenolics, flavonoids and antioxidant activity were found high in plant extracts mixtures than individual plants. Highest phenolics (580±1.12mg GAE/g), flavonoids (67.10±0.11mg CE/g) and antioxidant activity (IC50 0.109mg/ml) was observed with ratio 1:1:1:2 of plant mixture C, Cr, T, R. A weak linear positive correlation was found between antioxidant activity, total phenolic and flavonoid contents. A negative correlation was observed among IC50 value, total phenolics and flavonoid contents. Investigation through RP-HPLC revealed the presence of different potent phenolics in plants understudy. More antioxidant potential of extracts in combinations as compared to that of individual plants was clear corroboration of synergism. The ratio (1:1:1:2) of the studied plants in combination, that showed the highest free radical potential, was another expected better pharmacological prospect. This formulation can bring maximum relief against free radical-associated diseases.

In vitro antioxidant, hepatoprotective potential and chemical profiling of Syzygium aromaticum using HPLC and GC-MS.

The objective of the present study was to evaluate the in vitro antioxidant and hepatoprotective potential of Syzygium aromaticum (clove) against CCl4-induced hepatotoxicity using rat liver slice culture (LSC) model. Antioxidant activity in terms of DPPH radical scavenging activity and ferric reducing antioxidant power (FRAP) of different concentrations of S. aromaticum was in the range of 41.01-90.33% and 138.15-595.63 Fe (II) mg/mL, respectively. Plasmid pBR322 DNA protection activity was observed with all three concentrations of S. aromaticum against H2O2 induced oxidative damage, as no strand breaks were observed. Chemical profiling through HPLC confirmed the presence of six major phenolic acids and 13 volatile bioactive compounds were identified though GC-MS. Significant hepatoprotection (p<0.05) was observed in liver slice culture (LSC) as liver slices treated with various concentrations of S. aromaticum extract presented very low percentage cytotoxicity (7.35-16.16%) as compared to the CCl4 treated liver slices (75.58 %). The hepatoprotective potential of S. aromaticum may be due to the presence of bioactive components as confirmed by HPLC and GC-MS. The results of present study support the use of S. aromaticum in the formation of potential hepatoprotective drugs against various liver diseases.

Protection of DNA during oxidative stress and cytotoxic potential of Artemisia absinthium.

Medicinal plants are rich in secondary metabolites (alkoloids, glycosides, coumarins, flavonides, steroids, etc.) and considered to be more effective and a safer alternative source to manage a variety of diseases related to liver, heart and kidney disordered. This study determines in vitro antioxidant and in vivo toxicological profile including hemolytic, brine shrimp lethality and mutagenicity of aerial parts of Artemisia absinthium. DNA protection assay was performed on pUC19 plasmid vector using H(2)O(2) as oxidative agent. Total phenolic and flavonoid content were determined using colorimetric methods. Toxicity of the plant was evaluated by brine shrimp lethality, hemolytic and mutagenic activity. DNA protection assay of the plant showed concentration dependent protective effect and at concentration 10µL/mL revealed complete protective effect against H(2)O(2) induced DNA damage. Highest phenolic and flavonoid content was found to be 167.3 (mg GAE 100g DW(-1)) and 14 (mg CE 100g DW(-1)) respectively. Results showed that A. absinthium is potent against standard toxicological procedures, that indicates the presence of bioactive components in the plant and possess antioxidant activity that protects DNA against H(2)O(2) induced oxidative damage. Thus the results showed/support that A. absinthium provides significant health benefits.

Cardioprotective Potential of Polyphenolic Rich Green Combination in Catecholamine Induced Myocardial Necrosis in Rabbits.

The present study was designed to develop safer, effective, and viable cardioprotective herbal combination to control oxidative stress related cardiac ailments as new alternatives to synthetic drugs. The synergetic cardioprotective potential of herbal combination of four plants T. arjuna (T.A.), P. nigrum (P.N), C. grandiflorus (C), and C. oxyacantha (Cr) was assessed through curative and preventive mode of treatment. In preventive mode of treatment, the cardiac injury was induced with synthetic catecholamine (salbutamol) to pretreated rabbits with the proposed herbal combination for three weeks. In curative mode of treatment, cardiotoxicity/oxidative stress was induced in rabbits with salbutamol prior to treating them with plant mixture. Cardiac marker enzymes, lipids profile, and antioxidant enzymes as biomarker of cardiotoxicity were determined in experimental animals. Rabbits administrated with mere salbutamol showed a significant increase in cardiac marker enzymes and lipid profile and decrease in antioxidant enzymes as compared to normal control indicating cardiotoxicity and myocardial cell necrosis. However, pre- and postadministration of plant mixture appreciably restored the levels of all biomarkers. Histopathological examination confirmed that the said combination was safer cardioprotective product.

Gemmomodification: an emerging source of natural antioxidants from Silybum marianum.

Gemmomodification is a form of herbal medicine in which young freshly growing buds of plants are used. At germinating stage, plants metabolic activities are maximum and various nutrients, hormones enzymes and bioactive phytoconstituents are released and available at this stage. Plants may be promising source of natural antioxidants at growing stage. Oxidative stress leads to many chronic and degenerative diseases. Antioxidants are very essential for human body; they can protect the body from damage caused by free radical induced oxidative stress. This research project had been designed to investigate the antioxidant potential of gemmo modified and native (dry leaves) extract of Silybum marianum. Total phenolic contents was calculated by using Folin-Ciocalteu reagent and antioxidant potential was evaluated by using four radical assays (DPPH, ABTS, Super oxide and nitric oxide), reducing power assay and lipid peroxidation assay spectrophotometrically. Gemmo modified extract showed significantly higher (p<0.050) TPC (830 mg GAE/g of plant extract) as compared to native extract (800 mg GAE/g.) Results of this study revealed that gemmo modified extract demonstrated better antioxidant potential than natively used parts of Silybum marianum.