TPGS-modified Chitosan Nanoparticles of EGFR Inhibitor: Physicochemical and In vitro Evaluation against HepG2 Cell Lines.

BACKGROUND: Gefitinib (GFN) is an Epithelial Growth Factor Receptor (EGFR) inhibitor, and Food and Drug Administration (FDA) has approved medication to treat lung cancer. However, this investigation aimed to produce and characterize Gefitinib (GFN)-loaded chitosan and soy lecithin nanoparticles (NPs) modified with D-α-tocopheryl polyethylene glycol 1000 succinate mono ester (TPGS) and assess their therapeutic potential against HepG2 liver cell lines. METHODS: Chitosan, a cationic polymer with biocompatible and biodegradable properties, was combined with soy lecithin to develop the NPs loaded with GFN using a self-organizing ionic interaction methodology. RESULTS: The entrapment efficiency and drug loading were found to be 59.04±4.63 to 87.37±3.82% and 33.46±3.76 to 49.50±4.35%, respectively, and results indicated the encapsulation of GEN in NPs. The pH of the formulations was observed between 4.48-4.62. Additionally, all the prepared NPs showed the size and PDI range of 89.2±15.9 nm to 799.2±35.8 nm and 0.179±0.065 to 0.455±0.097, respectively. The FTIR bands in optimized formulation (GFN-NP1) indicated that the drug might be contained within the NP's core. The SEM photograph revealed the spherical shape of NPs. The kinetic release model demonstrated the combination of diffusion and erosion mechanisms. The IC50 value of GFN and GFN-NP1 formulation against the HepG2 cell lines were determined and found to be 63.22±3.36 µg/ml and 45.80±2.53 µg/ml, respectively. DAPI and PI staining agents were used to detect nuclear morphology. CONCLUSION: It was observed that the optimized GFN-NP1 formulation successfully internalized and inhibited the growth of HepG2 cells. Hence, it can be concluded that the prepared NPs can be a new therapeutic option for treating liver cancer.

Antimicrobial Peptides: The Production of Novel Peptide-Based Therapeutics in Plant Systems.

The increased prevalence of antibiotic resistance is alarming and has a significant impact on the economies of emerging and underdeveloped nations. The redundancy of antibiotic discovery platforms (ADPs) and injudicious use of conventional antibiotics has severely impacted millions, across the globe. Potent antimicrobials from biological sources have been extensively explored as a ray of hope to counter the growing menace of antibiotic resistance in the population. Antimicrobial peptides (AMPs) are gaining momentum as powerful antimicrobial therapies to combat drug-resistant bacterial strains. The tremendous therapeutic potential of natural and synthesized AMPs as novel and potent antimicrobials is highlighted by their unique mode of action, as exemplified by multiple research initiatives. Recent advances and developments in antimicrobial discovery and research have increased our understanding of the structure, characteristics, and function of AMPs; nevertheless, knowledge gaps still need to be addressed before these therapeutic options can be fully exploited. This thematic article provides a comprehensive insight into the potential of AMPs as potent arsenals to counter drug-resistant pathogens, a historical overview and recent advances, and their efficient production in plants, defining novel upcoming trends in drug discovery and research. The advances in synthetic biology and plant-based expression systems for AMP production have defined new paradigms in the efficient production of potent antimicrobials in plant systems, a prospective approach to countering drug-resistant pathogens.

In Silico Analysis of the Effect of Hydrastis canadensis on Controlling Breast Cancer.

Background and Objectives: Breast cancer is a significant type of cancer among women worldwide. Studies have reported the anti-carcinogenic activity of Hydrastis Canadensis (Goldenseal) in cancer cell lines. Hydrastis Canadensis could help eliminate toxic substances due to its anti-cancer, anti-inflammatory, and other properties. The design phase includes the identification of potential and effective molecules through modern computational techniques. Objective: This work aims to study Hydrastis Canadensis's effect in controlling hormone-independent breast cancer through in-silico analysis. Materials and Methods: The preliminary screening of reported phytochemicals includes biomolecular networking. Identifying functionally relevant phytochemicals and the respective target mutations/genes leads to selecting 3D proteins of the desired mutations being considered the target. Interaction studies have been conducted using docking. The kinetic and thermodynamic stability of complexes was studied through molecular dynamic simulation and MM-PBSA/GBSA analysis. Pharmacodynamic and pharmacokinetic features have been predicted. The mechanism-wise screening, functional enrichment, and interactional studies suggest that canadaline and Riboflavin effectively interact with the target proteins. Results: Hydrastis Canadensis has been identified as the effective formulation containing all these constituents. The phytoconstituents; Riboflavin and Canadensis showed good interaction with the targets of hormone-independent breast cancer. The complexes were found to be kinetically and thermodynamically stable. Conclusions: Hydrastis Canadensis has been identified as effective in controlling 'hormone-independent or basal-like breast cancer' followed by 'hormone-dependent breast cancer: Luminal A' and Luminal B.

Protective Effects of Fermented Houttuynia cordata Against UVA and H2O2-Induced Oxidative Stress in Human Skin Keratinocytes.

The biological activities of Houttuynia cordata (H. cordata) fermented with Aureobasidium pullulans (A. pullulans) was investigated for human skin keratinocyte-induced chemical and photo oxidations. In this research, H2O2/UVA-induced HaCaT cell lines were treated with H. cordata water/ethanol extracts (HCW/HCE) and fermented with A. pullulans water/ethanol extracts (HCFW/HCFE). A. pullulans fermented with H. cordata (HCFW) increased in 5.4-folds of total polyphenol (HCFW 46.89 mg GAE/extract g), and 2.3-folds in flavonoids (HCFW 53.80 mg GAE/extract g) compared with water extracts of H. cordata (HCW). Further, no significant cytotoxicity for HaCaT cells showed by all the extracts of H. cordata fermented with A. pullulans. HCFW extracts have significantly lowered inflammation factors such as COX-2 and Hsp70 proteins in oxidative stressed HaCaT cells induced by H2O2 and UVA treatments. All H. cordata extracts significantly downregulated gene expression involved in oxidative stress and inflammation factors, including IL-1ß, IL-6, COX-2, TNF-α, NF-κB, and MMP-1 in the H2O2/UVA-treated HaCaT cells. However, keratin-1 gene expression in the UVA-treated HaCaT cells was increased in twofolds by HCFW extracts. Further, A. pullulans fermented H. cordata extracts (HCFW/HCFE) reduced the genes involved in oxidative stresses more effectively than those of H. cordata extract only. Overall, the polyphenol-rich extracts of H. cordata fermented with A. pullulans showed synergistic protective effects for human epidermal keratinocytes to prevent photoaging and intrinsic aging by anti-oxidation and anti-inflammatory functions.

Anti-Inflammatory, Antioxidative, and Nitric Oxide-Scavenging Activities of a Quercetin Nanosuspension with Polyethylene Glycol in LPS-Induced RAW 264.7 Macrophages.

Quercetin (Qu) is a dietary antioxidant and a member of flavonoids in the plant polyphenol family. Qu has a high ability to scavenge reactive oxygen species (ROS) and reactive nitrogen species (RNS) molecules; hence, exhibiting beneficial effects in preventing obesity, diabetes, cancer, cardiovascular diseases, and inflammation. However, quercetin has low bioavailability due to poor water solubility, low absorption, and rapid excretion from the body. To address these issues, the usage of Qu nanosuspensions can improve physical stability, solubility, and pharmacokinetics. Therefore, we developed a Qu and polyethylene glycol nanosuspension (Qu-PEG NS) and confirmed its interaction by Fourier transform infrared analysis. Qu-PEG NS did not show cytotoxicity to HaCaT and RAW 264.7 cells. Furthermore, Qu-PEG NS effectively reduced the nitrogen oxide (NO) production in lipopolysaccharide (LPS)-induced inflammatory RAW 264.7 cells. Additionally, Qu-PEG NS effectively lowered the levels of COX-2, NF-κB p65, and IL-1ß in the LPS-induced inflammatory RAW 264.7 cells. Specifically, Qu-PEG NS exhibited anti-inflammatory properties by scavenging the ROS and RNS and mediated the inhibition of NF-κB signaling pathways. In addition, Qu-PEG NS had a high antioxidant effect and antibacterial activity against Escherichia coli and Bacillus cereus. Therefore, the developed novel nanosuspension showed comparable antioxidant, anti-inflammatory, and antibacterial functions and may also improve solubility and physical stability compared to raw quercetin.

Antioxidant and Antibacterial Profiling of Pomegranate-pericarp Extract Functionalized-zinc Oxide Nanocomposite.

With the advancement in green nanotechnology, considerable attention is being given to the synthesis of different kinds of nanomaterials for biological applications. In this study, zinc oxide nanocomposites (ZnO NPs) were synthesized using Punica granatum L. (Pomegranate) pericarp ethanolic extract (PE) by the chemical precipitation method. The prepared ZnO NPs showed a characteristic peak at 270 nm in the UV-Vis spectrophotometer and chemical bond stretching in the Fourier transforms infrared spectroscopy (FT-IR) spectra, indicated the formation of PE-functionalized zinc oxide nanocomposite (PE-ZnO NPs). The SEM results showed agglomerated PE-ZnO NPs of a spherical shape with an average size of 80-100 nm. Moreover, biological assessment of the PE-ZnO NPs revealed significant scavenging activity in DPPH (116.5%) and ABTS+ (95.2%) radical assay methods, and substantial antibacterial activity against Bacillus cereus, Bacillus licheniformis, and Escherichia coli. Furthermore, PE-ZnO NPs showed about 96.3% of cell viability for human HaCaT cells at the maximum concentration (100 µg/mL), marked as a reliable bioactive agent. Therefore, the developed PE-ZnO NPs were elucidated with substantial ROS scavenger and non-antibiotic antibacterial agent and hence, can be applied in respective biological applications.

Bioengineered gold nanoparticles using Cynodon dactylon extract and its cytotoxicity and antibacterial activities.

In this study, simple and green route approach was applied for the synthesis gold nanoparticles (AuNPs) containing an aqueous extract of Cynodon dactylon L. Pers., (C. dactylon). The synthesized AuNPs were characterized using spectral and microscopic analysis. The changes in the color pattern were observed upon synthesis by UV-vis spectrophotometer with a peak of 530 nm. The FT-IR, XRD, SEM, and TEM were used to analyze the crystal nature and morphology of the green synthesized AuNPs. The C. dactylon-loaded AuNPs in different concentrations (0.625-100 µg/ml) were used to assess cytotoxicity activity against MCF-7 cell line and where the IC50 was found to be 31.34 µg/ml by MTT assay. The C. dactylon-AuNPs were significantly increased reactive oxygen species (ROS) generation, DNA fragmentation, and mitochondrial membrane changes observed by dichlorodihydroflurescenin diacetate (DCFH-DA), 4',6-diamidino-2-phenylindole (DAPI), Rhodamine-123, and acridine orange (AO)/ethidium bromide (EtBr) staining assay. Besides the microbial study revealed that C. dactylon-AuNPs exhibited significant antibacterial activity against clinically isolated pathogenic bacteria such as Enterobacter cloacae, Staphylococus Haemolytics, Staphylococcus petrasii subsp. Pragensis and Bacillus cereus with a zone of inhibition 13, 12, 13 and 12 mm, respectively. It could be concluded that C. dactylon has the ability to be involved in the biosynthesis of AuNPs, and the pharmacological studies proved the promising cytotoxic effect on MCF-7 cell line and pathogenic bacterial species.

Biochemical and molecular aspects of 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis: a review.

1,2-dimethylhydrazine (DMH) is a member in the class of hydrazines, strong DNA alkylating agent, naturally present in cycads. DMH is widely used as a carcinogen to induce colon cancer in animal models. Exploration of DMH-induced colon carcinogenesis in rodent models provides the knowledge to perceive the biochemical, molecular, and histological mechanisms of different stages of colon carcinogenesis. The procarcinogen DMH, after a series of metabolic reactions, finally reaches the colon, there produces the ultimate carcinogen and reactive oxygen species (ROS), which further alkylate the DNA and initiate the development of colon carcinogenesis. The preneolpastic lesions and histopathological observations of DMH-induced colon tumors may provide typical understanding about the disease in rodents and humans. In addition, this review discusses about the action of biotransformation and antioxidant enzymes involved in DMH intoxication. This understanding is essential to accurately identify and interpret alterations that occur in the colonic mucosa when evaluating natural or pharmacological compounds in DMH-induced animal colon carcinogenesis.

Guava leaves extract ameliorates STZ induced diabetes mellitus via activation of PI3K/AKT signaling in skeletal muscle of rats.

Blood glucose homeostasis and insulin signaling pathway regulation take a vital role in the management of diabetes mellitus. Our present was designed to explore the mechanism of the blood homeostasis, regulation of oxidative stress and insulin signaling pathway by guava leaf extract (GLE). Diabetes mellitus was induced in male albino Wistar by streptozotocin (STZ) (Single dose-40 mg/kg b.w.). As an extension STZ rats received GLE (GLE; 200 mg/kg b.w). At the end of the study the lipid peroxidation products, antioxidants, insulin signaling genes were analyzed. Treatment with GLE resulted in decreased plasma and skeletal muscle lipid peroxidation markers, increased antioxidants, and improved insulin signaling genes. GLE treatment helps to maintain blood homeostasis alleviates oxidative stress and regulates the insulin signaling genes in skeletal muscle. Overall the results suggest GLE treatment regulates blood glucose, inhibits oxidative stress, and importantly it regulates insulin signaling pathway genes in skeletal muscle. Further studies on the GLE role in other important pathways can add additional strength to the claim that GLE is a strong anti-diabetic candidate.

Phloretin loaded chitosan nanoparticles enhance the antioxidants and apoptotic mechanisms in DMBA induced experimental carcinogenesis.

The main aim of this study was to investigate the effects of phloretin loaded chitosan nanoparticles (PhCsNPs) on 7,12-dimethylbenz[a]anthracene (DMBA) induced experimental cancer in hamsters. Oral squamous cell carcinoma (OSCC) was induced in male golden Syrian hamsters by painting with 0.5% DMBA three times a week for 14 weeks. Varying concentration of PhCsNPs (5, 10, and 20 mg/kg b.wt.) was orally administered on alternative days to evaluate the optimum dose. The experiment design was terminated at the end of the 14th week. The development of OSCC was confirmed by histopathological and biochemical analysis (lipid peroxidation, antioxidant profile, and detoxification enzymes) in plasma, erythrocyte, buccal, and liver tissues. Significant increases in oxidation and lipid peroxidation were noticed in DMBA-painted hamsters. Oral administration of PhCsNPs in various doses on alternate days reversed the deleterious effects induced by DMBA. In addition, immunoblot analyses of PhCsNPs treatment enhanced the release of Bcl-2 associated X protein (Bax), cytochrome c, caspase-3, 9 and suppressed the B-cell lymphoma 2 (Bcl-2) expression, which the use of PhCsNPs for mitochondrial-mediated apoptosis. These findings suggest biofabricated PhCsNPs may act as a potent antioxidant and anti-carcinogenic in DMBA induced oral cancer in experimental animals.

Phloretin loaded chitosan nanoparticles augments the pH-dependent mitochondrial-mediated intrinsic apoptosis in human oral cancer cells.

The aim of the present investigation is to explore the innovative platform for the synthesis of plant-based nanoparticles, which contain biocompatible and biodegradable carrier of chitosan loaded with phloretin hydrophobic phytochemical applied as a stable anticancer agent. Treatment of cancer uses chemotherapeutic drugs as the cells are resistant to other drugs. However, the usage of therapeutic drug is limited by its poor solubility and low bioavailability. To overcome this problem, we fabricated the phloretin loaded chitosan nanoparticles (PhCsNPs) and physicochemical properties of PhCsNPs were characterized by FTIR, XRD, DLS, SEM and TEM. The findings indicated that the synthesized PhCsNPs were spherical and homogeneous in shape with the size distribution of 80-100 nm and exhibited stability in ultimate drug releasing profile. Further, we substantiated the anticancer efficiency of PhCsNPs through bio-assessment, such as cytotoxicity measurement, intracellular ROS, mitochondrial dysfunction, lipid peroxidation measurement, antioxidants status, apoptotic associated gene expression profile and cell cycle analysis in human oral cancer cell lines. The findings suggested that PhCsNPs augmented the mitochondrial-mediated apoptotic mechanism through the stimulation of oxidative stress, depletion of cellular antioxidants and cell cycle arrest. Our data suggested that PhCsNPs could be used as an efficient therapeutic agent for the treatment of oral cancer.

Guava leaf inhibits hepatic gluconeogenesis and increases glycogen synthesis via AMPK/ACC signaling pathways in streptozotocin-induced diabetic rats.

Psidium guajava (PG) is a short shrub or tree cultivated in tropical and subtropical regions around the world. The leaf extract of PG (guava leaf) has been used historically to cure many ailments. However, mechanisms of action of guava leaf in treating diabetes are not fully understood. Effects and underlying mechanisms of guava leaf on gluconeogenesis and glycogenesis in hepatocytes, insulin signaling proteins, liver function markers, and lipid profile in streptozotocin (STZ) injected diabetic Wistar rats were investigated within the current study. PG was given orally at the dose of 100, 200, and 400 mg/kg b.w to diabetic rats for the period of 45 days. The results reveal that oral administration of PG (200 mg/kg b.w) has considerably raised the levels of insulin, glycogen, hexokinase, glucose-6-phosphatase dehydrogenase and significant (p < 0.05) belittled hepatic markers, gluconeogenic enzymes, and OGTT fasting blood glucose levels. OGTT has shown least statistical significance between the group 5 (200 mg/kg b.w) and group 6 and vital difference between group 5 and group 4 (400 mg/kg). PG has attenuated the triglycerides, total cholesterol, phospholipids, free fatty acid, and LDL levels and raised HDL levels. PG considerably (p < 0.05) activated IRS-1, IRS-2, Akt, p-Akt, PI3K, GLUT2, AMPK, p-AMPK, and p-ACC, which are the key effector molecules of the PI3K/Akt pathway in STZ rats. The results of our study specify that treatment with PG ameliorated glucose-metabolism and lipid profile in STZ evoked diabetic rats; the rationale ought to be the activation of PI3K/Akt, phosphorylation of AMPK pathway in liver and therefore has beneficial anti-diabetic activity.

Guava Leaf Extract Diminishes Hyperglycemia and Oxidative Stress, Prevents ß-Cell Death, Inhibits Inflammation, and Regulates NF-kB Signaling Pathway in STZ Induced Diabetic Rats.

Traditional Chinese medication has been utilized by Chinese medical practitioners to treat the varied symptoms of diabetes mellitus (DM). Notably, guava leaf has been used to treat diabetes in Asia. Our present study has been designed to analyze the action of guava leaf extract (GLE) at the molecular level in treating DM. A low dose of streptozotocin (STZ) was used to induce experimental diabetes in animals. Rats were treated with GLE at different concentrations (100, 200, and 400 mg/kg b.w.). The standard drug glibenclamide (GB) (600 µg/kg b.w.) was used for comparison. The diabetic rats showed a reduced level of insulin, accompanied by exaggerated levels of blood glucose, lipid peroxidation product, and augmented expressions of inflammatory cytokines, and showed reduced levels of antioxidants compared to the control rats. Supplementation with GLE counteracted the consequences of STZ. It suppresses the oxidative stress and inhibits the state of inflammation and the results are almost similar to that of standard drug group (GB group 5). Our present research, therefore, provides useful data concerning guava leaf extract by a thorough assessment in diabetes management. Being a natural product, additional analysis on GLE can shed light on finding effective phytochemicals within the field of diabetes mellitus.

7, 8-Dihydroxycoumarin (daphnetin) protects INS-1 pancreatic ß-cells against streptozotocin-induced apoptosis.

BACKGROUND AND OBJECTIVE: Daphnetin (7, 8-dihydroxycoumarin), a natural coumarin compound, is known to exhibit antioxidant and anti-inflammatrory effects. However, the underlying mechanisms of its anti-apoptotic and antidiabetic effects yet not been examined. Therefore, the present work studied the anti-apoptotic and anti-diabetic effects of daphnetin by in vitro experiments. METHODS: The rat insulinoma (INS-1) cells were pre-treated with daphnetin at different concentrations (1, 10, 20 and 40µM) for 24h followed by exposition to streptozotocin (STZ) (3mM) for 12h. Effects of daphnetin and STZ on INS-1 cells were determined by MTT assay, glucose stimulated insulin secretion (GSIS) assay, lipid peroxidation, antioxidant status (SOD, CAT, GPx, and GST) Apoptosis staining (DAPI, Hoechst 33342, AO/EB and ROS) was performed by fluorescence microscopy, and Bcl-2, Bax and NF-κB protein expression was detected by Western blotting. RESULTS: MTT assay indicated that the viability of INS-1 cells was significantly reduced with exposure to STZ for 12h as compared to control cells, while pre-treated with daphnetin for 24h resulted in a significant improvement of cell viability. The effects daphnetin treatment in INS-1 cells on insulin secretion was tested and results showed that the pre-treatment of daphnetin could improve GSIS. Further, daphnetin pre-treatment significantly reduced the levels of lipid peroxidation markers and also improved antioxidant enzymes' activities in STZ-induced INS-1 cells. Western blotting assay revealed that daphnetin could suppress apoptosis through up-regulation of anti-apoptotic Bcl-2 protein expression and the down-regulation of pro-apoptotic Bax and nuclear factor NF-κB protein levels. CONCLUSION: The results showed that daphnetin might be used in treating diabetes due to its insulin stimulating property and subsequent regulation of apoptotic pathway.

Antidiabetic Effects of Simple Phenolic Acids: A Comprehensive Review.

Diabetes mellitus (DM) has become a major public health threat across the globe. Current antidiabetic therapies are based on synthetic drugs that very often have side effects. It has been widely acknowledged that diet plays an important role in the management of diabetes. Phenolic acids are widely found in daily foods such as fruits, vegetables, cereals, legumes, and wine and they provide biological, medicinal, and health properties. Simple phenolic acids have been shown to increase glucose uptake and glycogen synthesis, improve glucose and lipid profiles of certain diseases (obesity, cardiovascular diseases, DM, and its complication). The current review is an attempt to list out the antidiabetic effects of simple phenolic acids from medicinal plants and botanical foods.

Antidiabetic properties of dietary flavonoids: a cellular mechanism review.

BACKGROUND: Natural food products have been used for combating human diseases for thousands of years. Naturally occurring flavonoids including flavones, flavonols, flavanones, flavonols, isoflavones and anthocyanidins have been proposed as effective supplements for management and prevention of diabetes and its long-term complications based on in vitro and animal models. AIM: To summarize the roles of dietary flavonoids in diabetes management and their molecular mechanisms. FINDINGS: Tremendous studies have found that flavonoids originated from foods could improve glucose metabolism, lipid profile, regulating the hormones and enzymes in human body, further protecting human being from diseases like obesity, diabetes and their complications. CONCLUSION: In the current review, we summarize recent progress in understanding the biological action, mechanism and therapeutic potential of the dietary flavonoids and its subsequent clinical outcomes in the field of drug discovery in management of diabetes mellitus.