Recuperative potential of Indian medicinal plant compounds- a tool to encumber henipaviruses: an in -silico study.

Henipaviruses, highly fatal zoonotic viruses with mortality rates up to 100%, pose a significant threat to humans. Despite sporadic cases, including infections from Cedar, Langya, and Nipah Viruses, there are no established drugs or vaccines for treatment. This lack of specific medication led us to explore 57 non-toxic compounds from Indian Medicinal Plants, selected from 232 compounds, aiming to combat these viruses. Through in silico ADMET analyses, Three compounds-andrographolide, pterygospermin and Salidroside-stood out for their exceptional non-toxic properties. These compounds underwent in silico target prediction, molecular docking and dynamics with Cedar, Langya, and Nipah Virus proteins from the Protein Data Bank. Among them, Andrographolide displayed the most promising negative free energy scores and stability in Cedar Virus-Attachment G-Protein binding pockets. Pterygospermin and Salidroside showed efficacy against Langya and Nipah Virus target proteins throughout the simulation. These compounds not only exhibited antiviral properties but also demonstrated immunomodulatory, anti-inflammatory, and hepatoprotective effects by our in-silico studies. Their potential as treatments or preventive measures against henipaviral infections makes them promising candidates for further research and development. Supplementary Information: The online version contains supplementary material available at 10.1007/s40203-024-00236-x.

Monoamine oxidase mediated oxidative stress: a potential molecular and biochemical crux in the pathogenesis of obesity.

Obesity has become a global health concern with an increasing prevalence as years pass by but the researchers have not come to a consensus on the exact pathophysiological mechanism underlying this disease. In the past three decades, Monoamine Oxidases (MAO), has come into limelight for a possible involvement in orchestrating the genesis of obesity but the exact mechanism is not well elucidated. MAO is essentially an enzyme involved in the catabolism of neurotransmitters and other biogenic amines to form a corresponding aldehyde, hydrogen peroxide (H2O2) and ammonia. This review aims to highlight the repercussions of MAO's catabolic activity on the redox balance, carbohydrate metabolism and lipid metabolism of adipocytes which ultimately leads to obesity. The H2O2 produced by these enzymes seems to be the culprit causing oxidative stress in pre-adipocytes and goes on to mimic insulin's activity independent of its presence via the Protein Kinase B Pathway facilitating glucose influx. The H2O2 activates Sterol regulatory-element binding protein-1c and peroxisome proliferator activated receptor gamma crucial for encoding enzymes like fatty acid synthase, acetyl CoA carboxylase 1, Adenosine triphosphate-citrate lyase, phosphoenol pyruvate carboxykinase etc., which helps promoting lipogenesis at the same time inhibits lipolysis. More reactive oxygen species production occurs via NADPH Oxidases enzymes and is also able activate Nuclear Factor kappa B leading to inflammation in the adipocyte microenvironment. This chronic inflammation is the seed for insulin resistance.

Exploring the molecular interactions and binding affinity of resveratrol and calcitriol with RAGE and its intracellular proteins and kinases involved in colorectal cancer.

Colorectal cancer (CRC) burden is progressively increasing in young population due to dietary and lifestyle pattern. Advanced glycation end products (AGEs), one of the dietary compounds, form complex aggregates with proteins, lipids, and nucleic acids distorting their structure and function. AGE's pro-tumorigenic role is mediated through the receptor for AGEs (RAGE) triggering an array of signaling pathways. The current study aimed to target AGE-RAGE axis signaling proteins and kinases at multiple levels with calcitriol (CAL) and trans-resveratrol (RES) through in silico analysis using molecular docking (MD), molecular dynamic simulation(MDS), MM-PBSA analysis, and in vitro study. In silico analysis of CAL and RES showed significant binding affinity toward RAGE and its signaling proteins such as NF-kB, PI3K/AKT, ERK1/2, and PKC compared to its reference inhibitors through better hydrogen, hydrophobic, pi-pi stacking interactions. MD and MDS studies have revealed stable and compact protein-ligand complexes. Binding free energies of protein-ligand complex were estimated using MM/PBSA analysis thatprovided an assessment of overall interacting free energies of complexes and revealed the presence of low binding energy within the active site. Furthermore, in the in vitro study, methylglyoxal (MG), an AGE-precursor showed a proliferative effect on HCT116, however, CAL and RES showed an inhibitory effect against MG induced effect with an IC50 value of 51 nM and 110 µM respectively. Thus, the study suggests the possible target binding sites of AGE-RAGE signaling proteins and kinases with CAL and RES, thereby exploiting it for developing CAL with RES as adjuvant therapy along with chemo drug for CRC.Communicated by Ramaswamy H. Sarma.

Sinapic acid prevents adipogenesis by regulating transcription factors and exerts an anti-ROS effect by modifying the intracellular anti-oxidant system in 3T3-L1 adipocytes.

Objectives: In this study, we tested the hypothesis that sinapic acid (SA), a naturally occurring hydroxycinnamic acid found in vegetables, cereal grains, and oilseed crops with various biological activities suppresses adipogenesis in 3T3-L1 adipocytes by down-regulating adipogenesis transcription factor. Materials and Methods: 3T3-L1 adipocytes were treated with SA and evaluated by Oil Red O staining, triglyceride estimation, lipolysis, and reverse transcription-polymerase chain reaction. 3T3-L1 adipocytes were treated with various concentrations of SA (100 to 1000 µmol) during differentiation. Results: SA prevented an increase in adipocytes by reducing preadipocyte clonal expansion. ORO staining analyses revealed that SA reduced cytoplasmic lipid droplet accumulation in 3T3-L1 by 57% at the highest concentration of 1000 µmol without affecting cell viability. Furthermore, SA down-regulated the expression of peroxisome proliferator-activated receptor-gamma, CCAAT/enhancer-binding protein alpha, sterol regulatory element-binding protein 1c, and fatty acid synthase. ROS generated during adipogenesis was also attenuated by SA treatment by increasing antioxidant enzymes superoxide dismutase, catalase, and the cellular antioxidant glutathione. SA demonstrated no in vivo toxicity in the Drosophila melanogaster model. Conclusion: These results suggest that SA exerts anti-oxidant and anti-adipogenic effects and could be used as a functional nutraceutical ingredient in combatting obesity-related diseases.

Inhibition of palmitic acid induced adipogenesis by natural polyphenols in 3T3-L1 adipocytes.

Dietary free fatty acids induce preadipocyte differentiation in the presence of a hormonal cocktail in 3T3-L1 adipocytes. Plant polyphenols are curb adipocyte differentiation and protect from metabolic stress. In the present study, we examined the effects of the saturated fatty acid, palmitic acid (PA) in presence of flavonoids, chrysin (CR) and hesperidin (HD) and phenolic acid, syringic acid (SYA) and sinapic acid (SIA). Adipocytes were incubated for 10 d with 100 µmol of PA along with 10-100 µmol CR/HD and 100-1000 µmol SYA/SIA. PA induced clonal expansion of preadipocytes, differentiation and oxidative stress in 3T3-L1 cells following 10 d of differentiation. Adipocytes treated with PA exhibited an increase of 300% in clonal population, 110% lipid and 172% reactive oxygen species accumulation. But treatment with CR, HD, SYA and SIA in the presence of PA concentration-dependent effect was observed. Concentrations of CR/HD and SYA/SIA inhibited PA-induced mRNA expression of PPARγ, C/EBPα, SREBP-1c, FAS and NOX4. Moreover, CR, HD, SYA and SIA did not exhibit toxicity in Drosophila DNA. In summary, these results suggest that dietary fatty acids act directly on adipocytes and addition of CR, HD, SYA and SIA resulted in reduction of PA-induced negative effects on 3T3-L1 adipocytes. HIGHLIGHTS: • Palmitic acid, the common dietary free fatty acid, is known to induce adipogenesis in 3T3-L1 adipocytes. • Treatment of differentiating adipocytes with flavonoids and phenolic acids reduced palmitic acid-induced clonal expansion of preadipocytes. • Phytocompounds reduced lipid accumulation and triglyceride production as well as ROS accumulation. • Thus, the phytocompounds showed effective anti-adipogenic activity even in palmitic acid challenged environment in adipocytes.

Enhanced Inhibition of Adipogenesis by Chrysin via Modification in Redox Balance, Lipogenesis, and Transcription Factors in 3T3-L1 Adipocytes in Comparison with Hesperidin.

OBJECTIVE: The present study was conducted to elucidate the in-vitro anti-oxidant and anti-adipogenic effect of the flavone, chrysin in comparison with the citrus bioflavonoid, hesperidin during adipogenic differentiation in 3T3-L1 mouse preadipocytes. METHODS: The effect of chrysin and hesperidin on adipogenic differentiation was evaluated using Oil red-O staining, triglyceride estimation, free glycerol release, and ROS accumulation. The expression of adipogenesis-related genes was evaluated in real time-polymerase chain reaction. RESULTS: 50 µmol chrysin or hesperidin did not affect the cell viability of 3T3-L1 preadipocytes and adipocytes, but significantly reduced preadipocyte clonal population, accumulation of intracellular lipid and ROS and consequently increased lipolysis and antioxidant enzyme defence. It also decreased the expression of major adipogenic transcription factors, CCAAT/enhancer-binding protein-ß, peroxisome proliferator activated receptor-γ, sterol regulatory element binding protein 1c, fatty acid synthase and hormone sensitive lipase. CONCLUSION(S): Herein we have indicated, for the first time, the effective anti-adipogenic mechanism of chrysin by down-regulating adipogenesis, lipogenesis and ROS and up-regulating lipolysis and antioxidant enzyme in differentiated 3T3-L1 adipocytes. As a nutritional bioflavonoid, chrysin with its more effective inhibition on adipogenesis than hesperidin has the potential to be developed as an anti-adipogenic nutraceutical agent.

Attenuation of tacrolimus induced oxidative stress, mitochondrial damage, and cell cycle arrest by Boerhavia diffusa root fraction in mdck cell lines.

OBJECTIVES: The protective effect of ethyl acetate fraction (EAF) of Boerhavia diffusa roots against Tacrolimus (TAC) induced nephrotoxicity was studied using MDCK cell lines. MATERIALS AND METHODS: Ethanolic root extract of B. diffusa was fractionated using the liquid-liquid partition method. The cytotoxic effect of TAC and protective effect of EAF co-treatment were studied in MDCK cell lines by measuring ROS, LPO, and NO levels; collagen accumulation, effect on mitochondrial membrane integrity and cell cycle analysis were studied. The active component in EAF was quantified by HPLC analysis. RESULTS: TAC induced toxicity, leading to apoptosis and necrosis, was significantly reduced (P<0.001) in EAF co-treatment, with reversal of cell cycle arrest and reduced cell population at sub G0/G1 phase. Further, ROS (P<0.05), LPO and NO (P<0.001), were significantly reduced with EAF co-treatment compared with TAC individually treated cells. TAC induced mitochondrial membrane integrity loss was found to be significantly reduced in co-treated cells, as measured by rhodamine123 (P<0.05) and translocation of cytochrome c (P<0.001) from nucleus to cytoplasm, and caspase 3 release (P<0.001). The same was confirmed through annexin-FITC and PI staining (P<0.05) with reduced apoptotic and necrotic death in co-treated population. Interestingly, EAF co-treatment decreased collagen accumulation (P<0.001) with significant increase in the cell survival of tubular epithelial cells. HPLC analysis showed the presence of Quercetin (87.5 mg/g) in EAF, which may be responsible for the nephroprotective role. CONCLUSION: Thus, these results provide sound evidence that EAF may be an effective adjuvant therapy to prevent nephrotoxicity induced by TAC.

3,5-Dimethoxy-4-benzoic acid (syringic acid) a natural phenolic acid reduces reactive oxygen species in differentiated 3T3-L1 adipocytes.

Preadipocytes under nutrient excess mature to lipid-laden adipocytes that are hotspots for generation of reactive oxygen species (ROS) imbalance and oxidative stress. Syringic acid (SA), a natural phenolic acid, was evaluated for its in vitro antioxidant and ROS modulation during in matured 3T3-L1 adipocytes. Following 10 d, the SA-treated adipocytes were evaluated for the levels of glutathione (GSH) and antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT). The levels of peroxides in mature adipocytes were estimated using dichlorofluorescein (DCF) cleavage fluorescence. The level of NADPH oxidase 4 (NOX4) expression was also investigated following 10-d differentiation period. SA significantly improved the levels of GSH, SOD, and CAT in matured adipocytes. Reduction in ROS production levels was also witnessed by decrease in DCF cleavage. SA showed concentration-dependent inhibition of NOX4 by day 7 of adipogenesis when compared with differentiated and undifferentiated cells. Moreover, SA exhibited effective antioxidant and anti-radical scavenging activity. These results suggest that SA in addition to inhibiting adipogenesis can strongly reduce ROS stress in mature adipocytes by upregulating levels of intracellular antioxidants and decreasing levels of NOX4 in 3T3-L1 adipocytes.

Impact of Advanced Glycation End products (AGEs) and its receptor (RAGE) on cancer metabolic signaling pathways and its progression.

Cancer is a complex disease with a 5-10% hereditary base, but nutrition, lifestyle, and the environment we are exposed to influence 90-95% of cancers. Due to rapid westernization, the diet we consume is rich in advanced glycation end products (AGEs). AGEs are the heterogeneous group of compounds formed by non-enzymatic reactions between reducing sugars and amino groups of proteins, lipids, and nucleic acids. Its implication is confirmed in many chronic conditions such as diabetes, renal, cardiovascular diseases, and aging however its role in cancer development has been understudied. Cancer cells are continuously exposed to AGEs due to their increased production, owing to its high metabolic rate and aerobic glycolysis. AGEs accumulation led to glycative stress which in turn stimulates oxidative stress and inflammation, through its receptor known as receptor for advanced glycation end products (RAGE). RAGE mediates crosstalk between the tumour cells and its microenvironment components to induce hypoxia, mitochondrial dysfunction, endoplasmic reticulum stress, autophagy, epigenetic modification, and cancer stemness. This emphasizes AGEs as an essential driving factor in different aspects of cancer development, but the exact molecular mechanism has to be explored. Thus, this review gives an insight into the pathological role of AGEs at the bio-molecular level in the tumourigenesis and progression of cancer in terms of the tumour microenvironment, invasion, and metastasis. Further, the compiled clinical data relating to the AGE-RAGE axis associated with different cancers and its potential inhibitors have been discussed.

In silico docking of phytocompounds to identify potent inhibitors of signaling pathways involved in prostate cancer.

Androgens and androgen receptors (AR) are the master regulators in the development of prostate cancer. Majority of the patients show positive response to surgical or medical castration, while many patients show disease relapse after the treatment. Genomic profiling has proven that the deregulated PI3K, Ras/Raf, MAPK and EGFR signaling pathways confer survival and invasion advantage to the cancer cells. Thus, modulation of these interlinked growth pathways along with androgen ablation may provide attractive therapeutic benefits. The current research is focused to identify the inhibitors of these pathways with bacosides and Piperine. The quantitative estimation of bacosides enriched standard extract of Bacopa monnieri by HPTLC showed 59.38% of Bacoside A and various active compounds with anti-oxidant, anti-cancer, anti-microbial, anti-inflammatory properties were also analyzed by GC-MS analysis. The in-vitro cytotoxic study against PC3 cell lines showed dose-dependent effect of Piperine and the extract. Further, in silico docking has shown bacosides with significant molecular interactions and binding score with growth factor receptors such as EGFR, PI3K, Akt and ERK, whereas Piperine exhibited interactions with AR. Hence, a simultaneous downregulation of interlinked signaling pathways of growth factors and AR with bacosides and Piperine may produce effective cytotoxic potential against the androgen-independent prostate cancer. Further in-vitro and in-vivo experimental investigations are necessary to determine the ultimate therapeutic utility. Communicated by Ramaswamy H. Sarma.

Phytofabricated silver nanoparticles: Discovery of antibacterial targets against diabetic foot ulcer derived resistant bacterial isolates.

The present study selected the predominant multi antibiotic-resistant diabetic foot ulcer (DFU) derived bacterial isolates such as Pseudomonas aeruginosa (PA), Escherichia coli (EC), Staphylococcus aureus (SA) and Bacillus subtilis (BS) and evaluated their response against the well-characterized Aerva lanata (AL) reduced multiple phytochemicals fabricated silver nanoparticles (AL-AgNPs). The overnight culture of DFU isolates was processed and subjected to various studies such as antimicrobial activity, growth kinetics, biofilm disruption, reactive oxygen species (ROS), membrane leakage, membrane permeability, and damage and genotoxicity. The molecular docking of AL phytochemicals was also performed with bacterial enzyme DNA gyrase. Interestingly, AL-AgNPs were produced the remarkable antibacterial effect against the resistant DFU isolates, which was closely similar to the effect of AL-AgNPs observed against the reference strains. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AL-AgNPs against the DFU isolates were found to be 5-15 µg/mL and 10-20 µg/mL, respectively. The AL-AgNPs were depicted a concentration-dependent growth inhibition of DFU bacterial isolates. The MIC and MBC of AL-AgNPs were effectively destroyed the preformed biofilms of DFU isolates. Furthermore, the MBC of AL-AgNPs was displayed the increased intracellular ROS accumulation, membrane leakage, permeability and damage, and genotoxicity in the DFU isolates. Additionally, the in silico study revealed that the AL phytochemicals were fitted over the binding pocket of the DNA gyrase B subunit. The observed results were confirmed that the negative impacts of the AL-AgNPs at the level of the membrane and intracellular components of DFU isolates.

Cellular growth factors as prospective therapeutic targets for combination therapy in androgen independent prostate cancer (AIPC).

Cancer is the second leading cause of death worldwide, with prostate cancer, the second most commonly diagnosed cancer among men. Prostate cancer develops in the peripheral zone of the prostate gland, and the initial progression largely depends on androgens, the male reproductive hormone that regulates the growth and development of the prostate gland and testis. The currently available treatments for androgen dependent prostate cancer are, however, effective for a limited period, where the patients show disease relapse, and develop androgen-independent prostate cancer (AIPC). Studies have shown various intricate cellular processes such as, deregulation in multiple biochemical and signaling pathways, intra-tumoral androgen synthesis; AR over-expression and mutations and AR activation via alternative growth pathways are involved in progression of AIPC. The currently approved treatment strategies target a single cellular protein or pathway, where the cells slowly develop resistance and adapt to proliferate via other cellular pathways over a period of time. Therefore, an increased research aims to understand the efficacy of combination therapy, which targets multiple interlinked pathways responsible for acquisition of resistance and survival. The combination therapy is also shown to enhance efficacy as well as reduce toxicity of the drugs. Thus, the present review focuses on the signaling pathways involved in the progression of AIPC, comprising a heterogeneous population of cells and the advantages of combination therapy. Several clinical and pre-clinical studies on a variety of combination treatments have shown beneficial outcomes, yet further research is needed to understand the potential of combination therapy and its diverse strategies.

Syringic acid (4-hydroxy-3,5-dimethoxybenzoic acid) inhibits adipogenesis and promotes lipolysis in 3T3-L1 adipocytes.

Increase in adiposity is associated with obesity and obesity-related complications. Syringic acid (SA), a phenolic acid facilitates oxidation, polymerization, and condensation reactions, may function as an effective anti-adipogenic agent. This study examined whether SA inhibits adipogenesis in 3T3-L1 preadipocytes by inhibiting differentiation and controlling lipid accumulation. Ten days of SA treatment suppressed differentiation of 3T3-L1 preadipocyte to mature adipocytes in a concentration-dependent manner as determined by Oil Red O staining. Suppression of adipogenic differentiation by SA was primarily from inhibition of mitotic clonal expansion during the early stages of differentiation. Treatment of adipocytes with SA reduced triglyceride production (lipogenesis) and increased glycerol release (lipolysis). The addition of SA to the adipocytes attenuated in vitro ROS production. These results indicate that SA inhibits the adipogenesis of 3T3-L1 preadipocytes, reduces lipid accumulation, and attenuates ROS accumulation, suggesting for the first time that SA has an anti-obesity effect in vitro.