Nitric Oxide Inhibition Research Articles

Cytotoxic and anti-inflammatory constituents from roots of Hypericum beanii and the antitumor potential under the view of cancer-related inflammation

Hypericum beanii, a traditional folk medicine plant, has been employed in the treatment of various inflammation-related diseases and has demonstrated promising potential as an herbal remedy for cancer. In this study, we isolated 29 compounds from the roots of H. beanii. We evaluated their cytotoxic effects on five human cancer cell lines, which revealed that the ethanol extract, along with compounds 4 and 14, exhibited significant cytotoxic activity. Additionally, we assessed their anti-inflammatory properties by measuring the inhibition of nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages. Our findings showed that the ethanol extract (IC50 = 7.41 ± 0.38 μg/mL), compound 4 (IC50 = 7.82 ± 0.42 μM), and compound 14 (IC50 = 3.05 ± 0.06 μM) displayed substantial anti-inflammatory activity. ELISA assays and qPCR analysis revealed that compounds 4 and 14 may exert their anti-inflammatory and antitumor effects by inhibiting the expression of iNOS, TNF-α, IL-1β, and IL-6 mRNA, shedding light on their role in cancer-related inflammation.

Abstract 15172: P21 Activated Kinase 2 Mediates Cardioprotection in Heart Failure With Preserved Ejection Fraction by Modulating Endoplasmic Reticulum Stress Response

Introduction: Heart failure with preserved ejection fraction (HFpEF) is a type of heart failure characterised by diastolic dysfunction but normal systolic function. Endoplasmic reticulum (ER) stress has been implicated in the pathophysiology of HFpEF. Previous studies have reported that P21 activated kinase 2 (Pak2) confers cardioprotection against the endoplasmic reticulum stress response. Hypothesis: Pak2 protein can potentially alleviate the adverse effects of maladaptive ER stress in HFpEF. Methods: The HFpEF model was developed by feeding C57/BL6N male mice with a high-fat diet (HFD) and a constitutive nitric oxide inhibitor, L-NAME, in drinking water for 15 weeks. Cardiac function was assessed by echocardiography and hemodynamics. Heart and lung structure was detected by H&E staining and Masson staining. Electron microscopy, immunoblotting, and RNA sequencing were used to investigate the ER structure and function. Cardiomyocyte-specific Pak2 knockout (Pak2 CKO ) mice and adeno-associated virus (AAV) 9 for Pak2 overexpression were used to investigate the role of Pak2 in the heart. Results: After 15 weeks of feeding, mice on the HFD+L-NAME diet developed an HFpEF phenotype, as indicated by cardiac hypertrophy, impaired diastolic function but preserved systolic function, and pulmonary remodelling. The ER structure and the IRE1/XBP1s pathway were impaired in the cardiomyocytes of HFD+L-NAME mice. Pak2CKO mice exhibited the HFpEF phenotype at 10 weeks. In addition, Pak2 overexpression prevented diastolic dysfunction after 15 weeks and preserved the activation of the IRE1/XBP1s pathway under stress conditions. Conclusions: HFD + L-NAME diet for 15 weeks recapitulated the features of HFpEF in mice. Pak2 plays a protective role in HFpEF progression by modulating the cardiomyocyte ER stress response through the IRE1/XBP1s signaling pathway.

Bioactivity of Fractions and Pure Compounds from Jatropha cordata (Ortega) Müll. Arg. Bark Extracts.

Medicines for chronic inflammation can cause gastric ulcers and hepatic and renal issues. An alternative treatment for chronic inflammation is that of natural bioactive compounds, which present low side effects. Extracts of Jatropha cordata (Ortega) Müll. Arg. have been evaluated for their cytotoxicity and anti-inflammatory activity; however, testing pure compounds would be of greater interest. Campesteryl palmitate, n-heptyl ferulate, palmitic acid, and a mixture of sterols, i.e., brassicasterol, campesterol, β-sitosterol, and stigmasterol, were obtained from an ethyl acetate extract from J. cordata (Ortega) Müll. Arg. bark using column chromatography. The toxicity and in vitro anti-inflammatory activities were evaluated using RAW 264.7 murine macrophage cells. None of the products assessed exhibited toxicity. The sterol mixture exhibited greater anti-inflammatory activity than the positive control, and nitric oxide (NO) inhibition percentages were 37.97% and 41.68% at 22.5 μg/mL and 30 μg/mL, respectively. In addition, n-heptyl ferulate decreased NO by 30.61% at 30 μg/mL, while campesteryl palmitate did not show anti-inflammatory activity greater than the positive control. The mixture and n-heptyl ferulate showed NO inhibition; hence, we may conclude that these compounds have anti-inflammatory potential. Additionally, further research and clinical trials are needed to fully explore the therapeutic potential of these bioactive compounds and their efficacy in treating chronic inflammation.

Essential oils from Acronychia pedunculata (L.) Miq. in Vietnam: chemical composition, antimicrobial, cytotoxicity and nitric oxide inhibition activities

The chemical composition, antimicrobial, cytotoxic and anti-inflammatory activities of essential oils from different parts of Acronychia pedunculata, a medicinal plant native to Central Highlands of Vietnam, were investigated. Their chemical constituents were determined using GC-MS. The antimicrobial properties were assessed using a microdilution broth method, while the anti-inflammatory properties were measured based on their capacity to inhibit NO production in macrophage cells. Their cytotoxicity was tested using the sulforhodamine B assay on three human cancer cell lines. The GC-MS analysis revealed the presence mainly of caryophyllene oxide (35.3%) and spathulenol (31.9%) in the stem essential oil, caryophyllene oxide (51.3%), and linalool (21.0%) in the leaf essential oil, and α-pinene (21.4%), (E)-β-ocimene (16.5%), linalool (12.5%), and β-caryophyllene (13.0%) in the fruit essential oil. All three essential oils inhibited the growth of various bacteria and fungi, reduced NO production in LPS-induced RAW264.7 cells, and exhibited cytotoxicity against all three tested cancer cell lines. Notably, the fruit essential oil showed the most potent cytotoxicity among the essential oils, with IC50 values of 5.47 ± 0.21, 7.95 ± 0.35, and 9.22 ± 0.52 g/mL against SK-LU-1, MCF-7, and HepG2 cells, respectively. It also had a strong anti-inflammatory effect that was comparable to that of dexamethasone, with an IC50 value of NO production inhibition of 14.62 ± 0.62 mg/mL. These results suggest the potential of the essential oils from this plant to provide leads for the development of anti-inflammatory and anti-cancer agents.

The characteristics and biological activity enhancements of melatonin encapsulations for skin care product applications

Melatonin (MLT) exhibits antioxidant, ultraviolet protection, anti-inflammatory, and anti-aging properties. However, its effectiveness is limited by instability, a short half-life, and incompatible absorption. In this research, we encapsulated melatonin (MLT) in transfersomes (MT) and niosomes (MN) to enhance their properties and investigate their effects through in vitro cell assays using murine macrophages cells and human foreskin fibroblasts cells. The vesicle morphology, vesicle size, polydispersity index, zeta potential, entrapment efficiency (EE%), attenuated total reflectance-Fourier transform spectroscopy (ATR-FTIR) spectra, along with in vitro release, permeation profiles, and stability study were also evaluated. The results showed that both encapsulations displayed spherical morphology at the nanometric scale, their great physical stability and provided an EE% range of 58–78%. The MLT incorporation into the vesicle was confirmed by the ATR-FTIR spectra. Additionally, the encapsulation’ release profiles fitted with the Higuchi model, indicating controlled release of melatonin. Furthermore, MT showed greater permeability than MN and MS including melatonin deposition. In cell assays, MT exhibited significantly higher nitric oxide inhibition and stimulation of collagen compared to MN and MS. Therefore, MT demonstrated the highest possibility for anti-inflammatory and collagen-stimulating activities that could be applied in pharmaceutical or anti-aging cosmetic products.

Virtual screening and invitro evaluation of cyclooxygenase inhibitors from Tinospora cordifolia using the machine learning tool

Tinospora cordifolia has a variety of compounds, and some of these compounds may have anti-inflammatory and antioxidant properties. In the present study, we identified the compounds in the leaf extract of T. cordifolia through Gas Chromatography-Mass Spectrometry (GC-MS) analysis and found the various metabolites. The compounds are screened virtually using a machine learning model, followed by molecular docking and simulation study to identify top-hit compounds as cyclooxygenase (COX) inhibitors. The molecular docking revealed that the compound 7,9-Di-tert-butyl-1-oxaspiro (4,5) deca-6,9-diene-2,8-dione (CID:545303) exhibited the lowest binding energies of −7.1 and −6.8 kcal/mol against COX 1 and COX 2 respectively. The interactions are favored by hydrogen bonding and hydrophobic interaction inside the binding pocket. The 100 ns MD simulation study for these compounds was performed to know the stability and found the RMSD around 2 Å and around 1.0 Å with minimal fluctuations indicating a stable complex throughout the simulation of 100 ns. Based on these findings, we proposed 7,9-Di-tertbutyl- 1-oxaspiro (4,5) deca-6,9-diene-2,8-dione could be used as a dual inhibitor of COX enzymes and a drug-like molecule for treating inflammation after evaluation of their biological properties. The methanolic extract of T. cordifolia was subjected to in vitro DPPH, ABTS, nitric oxide, anti-microbial, COX, and LOX inhibition activity. The results exhibited possible positive effects against the above activities. Communicated by Ramaswamy H. Sarma

Kaempferol protects against cardiovascular abnormalities induced by nitric oxide deficiency in rats by suppressing the TNF-α pathway

Kaempferol is a natural flavonoid compound that exhibits various pharmacological actions. However, there are few reports regarding the role of kaempferol in cardiovascular abnormalities. This study aimed to assess whether kaempferol could prevent cardiovascular malfunction and hypertrophy provoked by chronic inhibition of nitric oxide (NO) formation in rats. Rats (180–200 g) were treated daily with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) (40 mg/kg, in drinking water) for five weeks concomitant with kaempferol (oral administration) at a dose of 20 mg/kg or 40 mg/kg or lisinopril (5 mg/kg). Kaempferol partially prevented the progression of hypertension provoked by NO inhibition (p < 0.05). Left ventricular malfunction and hypertrophy present in hypertensive rats were alleviated by concurrent administration of kaempferol (p < 0.05). Furthermore, L-NAME rats had increased sympathetic nerve-mediated vasoconstriction and decreased acetylcholine-induced vasorelaxation and aortic wall thickening, which were resolved by kaempferol treatment (p < 0.05). Kaempferol restored tissue superoxide formation, malondialdehyde, catalase activity, plasma nitric oxide metabolites, tumor necrosis factor-alpha (TNF-α) and interleukin-6 in L-NAME rats (p < 0.05). Overexpression of tumor necrosis factor receptor 2 (TNFR2), phosphatidylinositol 3-kinases (PI3K), AKT serine/threonine kinase 1 (Akt1) and smad2/3 in heart tissue and upregulation of tumor necrosis factor receptor 1 (TNFR1), phosphorylated nuclear factor-kappaB (p–NF–κB) and transforming growth factor beta 1 (TGF-β1) in vascular tissue were suppressed by kaempferol (p < 0.05). In conclusion, kaempferol exerts antihypertensive, cardioprotective, antioxidant, and anti-inflammatory effects in NO-dependent hypertensive rats. The underlying mechanisms of kaempferol in preventing cardiovascular changes induced by L-NAME were due to the suppression of the TNF-α pathway.

Zebrafish nos2a benefits bacterial proliferation via suppressing ROS and inducing NO production to impair the expressions of inflammatory cytokines and antibacterial genes

The enzyme nitric oxide synthase 2 or inducible NOS (NOS2), reactive oxygen species (ROS) and nitric oxide (NO) are important participants in various inflammatory and immune responses. However, the functional significances of the correlations among piscine NOS2, ROS and NO during pathogen infection remain unclear. In teleost, there are two nos2 genes (nos2a and nos2b). It has been previously reported that zebrafish nos2a behaves as a classical inducible NOS, and nos2b exerts some functions similar to mammalian NOS3. In the present study, we reported the functional characterization of zebrafish nos2a during bacterial infection. We found that zebrafish nos2a promoted bacterial proliferation, accompanied by an increased susceptibility to Edwardsiella piscicida infection. The nagative regulation of zebrafish nos2a during E. piscicida infection was characterized by the impaired ROS levels, the induced NO production and the decreased expressions of proinflammatory cytokines, antibacterial genes and oxidant factors. Furthermore, although both inducing ROS and inhibiting NO production significantly inhibited bacterial proliferation, only inhibiting NO production but not inducing ROS significantly increased resistance to E. piscicida infection. More importantly, ROS supplementation and inhibition of NO completely abolished this detrimental consequence mediated by zebrafish nos2a during E. piscicida infection. All together, these results firstly demonstrate that the innate response mediated by zebrafish nos2a in promoting bacterial proliferation is dependent on the lower ROS level and higher NO production. The present study also reveals that inhibition of NO can be effective in the protection against E. piscicida infection.

Evaluation of Crocin Content and In Vitro Antioxidant and Anti-Glycation Activity of Different Saffron Extracts.

Crocin, a glycoside carotenoid that exhibits several health benefits, is mainly obtained from saffron (Crocus sativus L.), whose quality and content of phytochemicals can be strongly affected by environmental conditions. Therefore, in this work, the crocin content and in vitro antioxidant activity of saffron extracts obtained from three different varieties (Greek, Sicilian, and Iranian saffron) were assessed. Crocin content in saffron extracts was quantified via ultra-performance liquid chromatography coupled with mass spectrometry. The antioxidant activity of saffron extracts was evaluated using the oxygen radical absorbance capacity (ORAC) assay and nitric oxide (NO) radical scavenging test. The Maillard reaction was used to assess anti-glycation activity. Although the Sicilian and Iranian saffron extracts contained higher amounts of crocin (128 ± 6 ng/mL and 126 ± 4 ng/mL, respectively) compared to the Greek extracts (111 ± 2 ng/mL), ORAC values (50.9 ± 0.5) and % NO inhibition (35.2 ± 0.2) were higher for the Greek variety, which displayed a total phenolic content about two-fold greater than that of the other two extracts. Sicilian and Greek saffron had similar anti-glycation activities, while Iranian saffron was less effective. These results suggest that the antioxidant activity of saffron extracts could be ascribed to their naturally occurring complex mixture of phytochemicals, deserving further investigation as supplements to prevent pathological conditions induced by radical species.

Novel trans-caffeate hydrazide derivatives: synthesis, inhibition of nitric oxide (NO) production and molecular docking studies

Eight new caffeyl hydrazide derivatives (4a–4h) were synthesised via a convenient esterification of caffeic acid with some substituted aryl acid hydrazides. The synthesised caffeyl derivatives were evaluated for their inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 macrophages. The fluorobenzoylhydrazide derivatives 4f, 4 g and 4h were found to be the most powerful anti-inflammatory compounds with IC50 values ranging from 11.90 to 24.17 μM, which were more potent than the reference compound L-NMMA (IC50 32.8 μM). Additionally, synthesised compounds have been rationalised by using molecular docking studies which were performed in order to understand insights on the action mechanism of newly synthesised inhibitors against inflammatory mediator (iNOS). Obtained data indicate that compounds 4f, 4h, 4a and 4 g were observed to effectively bind to iNOS receptor with dock score values of −11.62, −10.81, −10.78 and −10.51 kcal/mol, respectively.

The Effects of Nitric Oxide Synthase Inhibition on Epinephrine-Induced Arrhythmia and Myocardial Damage

We have recently reported that methylene blue (MethyB) was able to inhibit epinephrine-induced arrhythmias and cardiac muscle injury. In this study, we investigated the effect of nitric oxide synthase inhibition by NG-nitro-L-arginine methyl ester (L-NAME) on cardiac arrhythmias, and myocardial damage induced by epinephrine in rats. Whether nitric oxide inhibition would affect the antiarrhythmic and cardiac protective actions of MethyB was also examined. L-NAME (40 mg/kg), L-arginine (200 mg/kg) + L-NAME, or MethyB (100 mg/kg) + L-NAME were given intraperitoneally (i.p.). Cardiac arrhythmia was then induced with intravenous (i.v.) injection of 10 μg/kg epinephrine. Results showed that epinephrine injection caused marked bradycardia (221.0 ± 1.37 vs. 409.4 ± 3.18 beats/min), shortened QTc interval (0.096 ± 0.0093 vs. 0.177 ± 0.0008 s), increased QRS duration (0.040 ± 0.0035 vs. 0.0185 ± 0.0002 s), decreased R wave amplitude (0.176 ± 0.0051 vs. 0.21 ± 0.0009 mv), ST segment height (-0.026 ± 0.007 vs. -0.002 ± 0.0005 mv), and induced ventricular extrasystoles. L-NAME given to untreated control rats resulted in a decrease in heart rate (288.2 ± 0.88 vs. 409.4 ± 3.18 beats/min), and increased R wave amplitude (0.436 ± 0.004 vs. 0.21 ± 0.0009 mv) compared to controls. L-NAME did not cause extrasystoles in untreated control rats but significantly increased the number of extrasystoles and duration of arrhythmia in the epinephrine-treated group. The administration of L-arginine (200 mg/kg, i.p.) to epinephrine plus L-NAME-treated rats resulted in increased heart rate and markedly decreased the number of extrasystoles and duration of arrhythmia. Methylene blue given at 100 mg/kg to rats treated with epinephrine and L-NAME caused a marked increase in heart rate. It also normalized QRS duration, prevented ST segment depression, markedly suppressed ventricular extrasystoles, and decreased the duration of arrhythmia compared with either epinephrine or L-NAME plus epinephrine-treated groups. Epinephrine injection caused disorganization, and necrosis of cardiac cells, interstitial hemorrhage, and cellular infiltrations. These changes were markedly improved by treatment with either L-NAME or L-NAME/MethyB. These results suggest that (i) inhibiting nitric oxide synthase by L-NAME increases epinephrine-induced arrhythmia which is inhibited by L-arginine or MethyB; (ii) either L-NAME alone or in combination with MethyB prevented cardiac muscle injury induced by epinephrine; (iii) L-NAME did not prevent the cardiac protective and antiarrhythmic actions of MethyB.

Polyoxygenated cyclohexene derivatives and other constituents of Uvaria rufa stem

Six undescribed compounds, uvarirufols D and E, (+)-uvarigranol B, (−)-uvarigranol E, 6-acetoxy-5-hydroxy-7-methoxyflavanone and cherrevenaphthalene D, along with twelve known compounds, including polyoxygenated cyclohexenes, flavonoids, and lignans, were isolated from the methanol extract of Uvaria rufa stems. Their structures were elucidated by spectroscopic analyses and the absolute configurations were determined using electronic circular dichroism. Several isolates were evaluated for cytotoxic, antitubercular and anti-inflammatory potentials. (−)-6-Acetylzeylenol showed moderate inhibitory activity against Mycobacterium tuberculosis, with MIC value of 47.10 μg/mL. Cherrevenaphthalene D exhibited weak antimycobacterial activity and potent inhibitory effect on lipopolysaccharide-induced nitric oxide (NO) production in RAW 264.7 cells (EC50 = 8.54 μM). 8-Hydroxy-5,7-dimethoxyflavanone displayed moderate level of NO inhibition (EC50 = 43.62 μM) with little cytotoxicity. The polyoxygenated cyclohexenes and lignans were inactive against HCT 116 and 22Rv1 cancer cells (IC50 > 100 μM).

Lithospermum erythrorhizon Siebold & Zucc. extract reduces the severity of endotoxin-induced uveitis

BackgroundUveitis is an inflammatory eye condition that threatens vision, and effective anti-inflammatory treatments with minimal side effects are necessary to treat uveitis. PurposeThis study aimed to investigate the effects of Lithospermum erythrorhizon Siebold & Zucc. against endotoxin-induced uveitis in rat and mouse models. MethodsEndotoxin-induced uveitis models of rats and mice were used to evaluate the effects of l. erythrorhizon treatment. Clinical inflammation scores and retinal thickness were assessed in the extract of l. erythrorhizon-treated rats. Histopathological examination revealed inflammatory cell infiltration into the ciliary body. Protein concentration, cellular infiltration, and prostaglandin-E2 levels were measured in the aqueous humor of the extract of l. erythrorhizon-treated rats. Protective effects of l. erythrorhizon on the anterior segment of the eye were examined in mice with endotoxin-induced uveitis. Additionally, we investigated the effect of l. erythrorhizon on the expression of pro-inflammatory cytokines [tumor necrosis factor alpha, interleukin-6, and interleukin-8] in lipopolysaccharide-stimulated THP1 human macrophages and examined the involvement of nuclear factor kappaB/activator protein 1 and interferon regulatory factor signaling pathways. Furthermore, three components of l. erythrorhizon were identified and assessed for their inhibitory effects on LPS-induced inflammation in RAW264.7 macrophage cells. ResultsTreatment of the extract of l. erythrorhizon significantly reduced clinical inflammation scores and retinal thickening in rats with endotoxin-induced uveitis. Histopathological examination revealed decreased inflammatory cell infiltration into the ciliary body. The extract of l. erythrorhizon effectively reduced the protein concentration, cellular infiltration, and PG-E2 levels in the aqueous humor of rats with endotoxin-induced uveitis. In mice with endotoxin-induced uveitis, the extract of l. erythrorhizon demonstrated a protective effect on the anterior segment of the eye by reducing inflammation and retinal thickening. The extract of l. erythrorhizon suppressed the expression of pro-inflammatory cytokines (tumor necrosis factor alpha, interleukin-6, and interleukin-8) in lipopolysaccharide-induced inflammation in THP1 human macrophages, by modulating nuclear factor kappaB/activator protein 1 and interferon regulatory factor signaling pathways. Moreover, shikonin, acetylshikonin, and β, β-dimethylacryloylshikonin showed dose-dependent inhibition of nitric oxide, tumor necrosis factor alpha and interleukin-6 production in RAW264.7 macrophage cells. ConclusionThe extract of l. erythrorhizon is a potential therapeutic agent for uveitis management. Administration of the extract of l. erythrorhizon led to reduced inflammation, retinal thickening, and inflammatory cell infiltration in rat and mouse models of uveitis. The compounds (shikonin, acetylshikonin, and β, β-dimethylacryloylshikonin) identified in this study played crucial roles in mediating the anti-inflammatory effects of l. erythrorhizon. These findings indicate that the extract of l. erythrorhizon and its constituent compounds are promising candidates for further research and development of novel treatment modalities for uveitis.

New Monoterpene Glycoside Paeoniflorin Derivatives as NO and IL-1β Inhibitors: Synthesis and Biological Evaluation.

Several monoterpene glycoside compounds were extracted from Paeonia lactiflora Pall. Among them, paeoniflorin, a water-soluble monoterpene glycoside found in the root of Paeonia lactiflora Pall, exhibits excellent antioxidant pharmacological functions. Initially, Sc(CF3SO3)3 was employed as the catalyst for paeoniflorin's dehydration and rearrangement reactions with alcohols. Subsequently, structural modifications were performed on paeoniflorin through a series of responses, including acetylation, deacetylation, and debenzoylation, ultimately yielding 46 monoterpene glycoside derivatives. The potential inhibitory effects on the pro-inflammatory mediators interleukin-1 beta (IL-1β) and nitric oxide (NO) were assessed in vitro. The results revealed that compounds 29 and 31 demonstrated notable inhibition of NO production, while eight derivatives (3, 8, 18, 20, 21, 29, 34, and 40) displayed substantial inhibitory effects on the secretion of IL-1β. Computational research was also undertaken to investigate the binding affinity of the ligands with the target proteins. Interactions between the proteins and substrates were elucidated, and corresponding binding energies were calculated accordingly. The findings of this study could provide valuable insights into the design and development of novel anti-inflammatory agents with enhanced pharmacological properties.

Isolation, purification and characterization of bioactive surfactant from the probiotic bacterium Lactobacillus casei NM512 isolated from infant microbiota

Background Lactobacillus casei NM512 was previously isolated from the feces of breast-fed infants and has great potential for probiotic treatment of diseases that are based on immune deregulation. These strains fulfill the main criteria required for safe human consumption and could therefore be useful in pharmaceutical and therapeutic preparations. However, further studies are needed to evaluate and decipher the in vivo activities of the isolated strains with regard to their immunomodulatory function. Objective This work aims to isolate, identify and characterize the bioactive surfactant component from the probiotic bacterium L. casei NM512. Materials and methods The isolation of the surfactant from L. casei NM512 was performed after incubation the strain for 72 h at 37°C with shaking at 120 rpm. The surfactant extracted from cells by acid precipitation. A combination of different methods (TLC, FT-IR, NMR, and GC mass) was used to analyze the composition of the isolated surfactant. The surfactant was also investigated for its antimicrobial activity using the well agar diffusion method, antioxidant ability targeting the DPPH free radical, Cytotoxicity ability using MTT assay, and anti- inflammatory activity was done by determination the inhibition of nitric oxide (NO) production in RAW 264.7 macrophages. Results and conclusion The obtained data show that the surfactant is a mixture of carbohydrates, lipids and proteins with glycolipopeptide nature that expressed remarkable antibacterial and antioxidant activities. Furthermore the isolated surfactant showed safety by keeping the cell viability and anti-inflammatory activity. Here we introduce a novel biosurfactant for medical and pharmaceutical purposes.

Gut microbiome of Crocodylus porosus and cellular stress: inhibition of nitric oxide, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells.

Crocodiles are renowned for their resilience and capacity to withstand environmental stressors, likely influenced by their unique gut microbiome. In this study, we determined whether selected gut bacteria of Crocodylus porosus exhibit anti-inflammatory effects in response to stress, by measuring nitric oxide release, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells. Using the Griess assay, the findings revealed that among several C. porosus gut bacterial isolates, the conditioned media containing the metabolites of two bacterial strains (CP27 and CP36) inhibited nitric oxide production significantly, in response to the positive control, i.e., taxol-treatment. Notably, CP27 and CP36 were more potent at reducing nitric oxide production than senloytic compounds (fisetin, quercetin). Using enzyme linked immunosorbent assays, the production of pro-inflammatory cytokines (IL-1β, TNF-α, PGE2), was markedly reduced by treatment with CP27 and CP36, in response to stress. Both CP27 and CP36 contain a plethora of metabolites to exact their effects [(3,4-dihydroxyphenylglycol, 5-methoxytryptophan, nifedipine, 4-chlorotestosterone-17-acetate, 3-phenoxypropionic acid, lactic acid,f-Honaucin A,l,l-Cyclo(leucylprolyl), 3-hydroxy-decanoic acid etc.], indicative of their potential in providing protection against cellular stress. Further high-throughput bioassay-guided testing of gut microbial metabolites from crocodiles, individually as well as in combination, together with the underlying molecular mechanisms, in vitro and in vivo will elucidate their value in the rational development of innovative therapies against cellular stress/gut dysbiosis.

In Vitro and In Vivo Anti-Aging Effect of Coffee Berry Nanoliposomes.

Encapsulation of bioactive compounds in the liposome system provides several advantages, such as enhancing the stability and lowering the toxicity of active compounds. Coffee berry extract (CBE) has previously been established to have in vitro anti-aging properties and to retard the aging of human skin. The purposes of this study were to encapsulate CBE in nanoliposomes and to assess its stability and in vitro anti-aging potential in human dermal fibroblasts (HDF), as well as in healthy human skin. In the HDF model, anti-aging potential was determined by nitric oxide (NO) and collagenase inhibition assays and a superoxide dismutase (SOD) activity assay, whereas in healthy human skin (in vivo), the skin elasticity and brightness were examined. First, liposomal CBE (L-CBE) was created with a particle size of 117.33 ± 2.91 nm, a polydispersity index (PDI) of 0.36 ± 0.03, and a zeta potential of -56.13 ± 1.87 mV. The percentages of encapsulation efficacy (%EE) and loading efficacy (%LE) were 71.26 ± 3.12% and 2.18 ± 0.18%, respectively. After undergoing a 12-week stability test, the L-CBE retained more phenolic content than the free CBE when stored at 4 °C, room temperature, and 45 °C. Compared to free CBE, the L-CBE demonstrated a more consistent, elevated, and prolonged release of phenolics from the lipid system. In human dermal fibroblasts, L-CBE showed lower toxicity, and at its maximum nontoxic concentration (10 mg/mL), it exhibited slightly higher anti-aging effects than CBE, including NO inhibition, enhanced SOD activity, and anti-collagenase activities. In clinical trials (30 volunteer subjects), none of the participants' skin was irritated when the L-CBE, the CBE, or base creams were applied. After 2 weeks of application, the L-CBE and CBE creams both demonstrated an improvement in skin elasticity and a reduction in melanin levels, and after 4 weeks, L-CBE cream showed a significantly greater improvement in skin elasticity and lightening. The results demonstrate that the encapsulation of the CBE in liposomal systems could increase its stability and skin penetration, reduce its toxicity, and maintain its anti-aging effect, which is powerful enough to be exploited in anti-aging and whitening agents for application in cosmetics and cosmeceuticals.

Ultraviolet B-induced increase of cyanidin-3-O-galactoside, quercetin-3-O-galactoside, and ursolic acid enhances antioxidant and anti-inflammatory activities in apples

Postharvest ultraviolet (UV) treatment is a promising method for enhancing the coloration of apple skin. However, the effects of this treatment on the bioactivities of apple skin components are not fully understood. This study aimed to evaluate changes in antioxidant and proinflammatory cytokine inhibition activities as well as the components of apple skin extracts following UV-B treatment. The results showed that UV-B treatment had minimal effects on the quality parameters of the fruit, such as firmness, soluble solids content, and titratable acidity, but it stimulated the accumulation of cyanidin-3-O-galactoside (C3gal). Quercetin-3-O-galactoside (Q3gal), quercetin-3-O-glucoside (Q3glu), and ursolic acid were the major components that exhibited significantly increased content in UV-B–irradiated apple skin extracts presenting 24.9, 7.9, and 11.1-fold higher content than dark treatment, respectively, with this study the first to reveal that UV-B treatment alters ursolic acid content in apples. UV-B treatment also increased both 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid antioxidant activities, although excessive UV-B intensity (>4 W m−2 s−1) decreased DPPH activity. Furthermore, UV-B–treated skin showed significantly suppressed nitric oxide and tumor necrosis factor-α (TNF-α) production but not interleukin-6 (IL-6) inhibition, which was not a specific effect of UV-B treatment as it is common effect of apple skin extracts. C3gal and Q3gal significantly inhibited IL-6 but not nitric oxide and TNF-α suppression. Overall, our findings indicate that UV-B intensity positively correlates with antioxidant activity resulting from the accumulation of C3gal, Q3gal, Q3glu, and ursolic acid. However, UV-B intensity did not correlate with anti-inflammatory effects, although the extract of UV-B–treated apple skin exhibited relatively strong inhibition of inflammation.

A Study on Antioxidant and Anti-inflammatory of Salvia microphylla Ethanol Extract

Purpose: This study was aimed at investigating the anti-inflammatory and antioxidant properties of &lt;ji&gt;Salvia microphylla (SM) extract (SME).Methods: Powdered SM was treated with ethanol and decocted to acquire SME. The antioxidant activities of SME were assessed by evaluating its total polyphenolic, and flavonoid , contents and performing 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. The anti-inflammatory activities of SME were verified by examining the inhibition of nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS) activated RAW 264.7 macrophages.Results: The total polyphenol and flavonoid contents in SME were 102.26±0.39 mg GAE/g and 377.42±0.77 mg QUE/g, respectively. The DPPH and ABTS assays revealed the antioxidant activities of SME by exhibiting their ability to scavenge radicals in a concentration-dependent manner (50-1000 μg/mL). The cell viability of SME exceeded 99% at 50-1000 μg/mL. The concentration-dependent reduction in the expression of the inflammation mediator iNOS and subsequent NO production indicated an approximately 100% inhibition efficacy at 600 μg/mL (0.00±1.34%). These results confirmed the potential of SME as a component of cosmeceuticals.Conclusion: Based on the findings of this study, SME is explicitly acknowledged for its applicability as a natural preservative possessing antioxidant and anti-inflammatory activities as well as a raw material for cosmeceuticals intended to alleviate skin irritation.

Nanoemulsion of Zanthoxylum rhetsa (Roxb.) DC: Formulation, release characteristics and its potential use for anti-inflammation applications

Nanoemulsions of Zanthoxylum rhetsa (Roxb.) DC fruit extract combined with eucalyptus oil (ZRNEs) were formulated using mixed surfactants at various Tween 80/Span 80 mass ratios of 20/80 (ZRNE-6), 50/50 (ZRNE-10), and 80/20 (ZRNE-13). These formulations had corresponding hydrophilic-lipophilic balance (HLB) values of 6, 10, and 13, respectively. Effect of the HLB on properties of the nanoemulsions were evaluated. Among these formulations, ZRNE-10, was found to be optimal for loading ZR extract with eucalyptus. It exhibited the smallest particle size of 402 ± 12 nm with a PDI of 0.41 ± 0.01, as analyzed by dynamic light scattering technique. ZRNE-10 showed the highest encapsulation efficiency for terpinen-4-ol at 83.42 ± 0.51%. It demonstrated a gradual release profile, releasing 30% of terpinen-4-ol within 48 h in a pH 7.4 PBS buffer at 37 °C. Interestingly, higher HLB values accelerated the release rate of terpinen-4-ol. The release kinetics of ZRNE-10 were best described by the Higuchi model (R2 = 0.903). ZRNE-10 displayed robust antioxidant activity (59.5 ± 1.6%) and effective inhibition of nitric oxide (76.9 ± 0.8%). It also exhibited low cytotoxicity on normal human skin fibroblasts (IC50 ≈ 3 mg/mL). Stability assessments revealed that all ZRNE formulations maintained their physical attributes and bioactivities even after one month of storage at 25 °C, and even at an elevated temperature of 45 °C. Notably, the antioxidant activities of ZRNEs surpassed those of unencapsulated ZR extract. Given their favorable anti-inflammatory properties, low cytotoxicity, and maintained efficacy, ZRNEs hold promise as potential ingredients for topical use.