Elastase Inhibitor Research Articles

Rubus urticifolius Compounds with Antioxidant Activity, and Inhibition Potential against Tyrosinase, Melanin, Hyaluronidase, Elastase, and Collagenase.

In our study, using chromatographic techniques, we isolated three bioactive compounds, which were structurally elucidated as (E)-2-(3-(3,4-dimethoxyphenyl)acrylamido)-N-methylbenzamide (1), 4-Hydroxyquinoline-2-carboxylic acid (2), and (E)-2-Cyano-3-(4-hydroxyphenyl)acrylic acid (3), using spectroscopic methods. The anti-melanogenic, anti-inflammatory, antioxidant, and anti-aging properties were evaluated in vitro by measuring the activity of pharmacological targets including tyrosinase, melanin, NF-κB, hyaluronidase, elastase, collagenase, and Nrf2. Our results show that compound 1 is the most active with IC50 values of 14.19 μM (tyrosinase inhibition), 22.24 μM (melanin inhibition), 9.82-12.72 μM (NF-κB inhibition), 79.71 μM (hyaluronidase inhibition), 80.13 μM (elastase inhibition), 76.59 μM (collagenase inhibition), and 116-385 nM (Nrf2 activation) in the THP-1, HEK001, WS1, and HMCB cells. These findings underscore the promising profiles of the aqueous extract of R. urticifolius at safe cytotoxic concentrations. Additionally, we report, for the first time, the isolation and characterisation of these nitrogenous compounds in the R. urticifolius species. Finally, compound 1, isolated from R. urticifolius, is a promising candidate for the development of more effective and safer compounds for diseases related to skin pigmentation, protection against inflammation, and oxidative stress.

Plectranthus scutellarioides (L.) R.Br. Leaf Extract as Sunscreen, Skin Lightening, and Antiaging

Plectranthus scutellaroides is a plant with attractive colored leaves. It contains secondary metabolite compounds such as flavonoids and phenolics, which provide skin protection against ultraviolet rays from the sun. The research aims to investigate the activity of P. scutellarioides leaf extract from various variants and different solvents on sunscreen, skin lightening, and anti aging activities. The best extract is formulated in gel dosage form. Each P. scutellaroides leaf was extracted using ethanol, ethyl acetate, and hexane as solvents. The obtained extract was assessed for Sun Protecting Factor (SPF), UVA Protection grade (PA), antioxidant activity, and inhibition of tyrosinase, collagenase, elastase, and hyaluronidase. Determination was conducted in vitro using spectrophotometric methods. The highest sunscreen activity was observed in the ethanol extract Va (Va-ET), which exhibited SPF and PA values of 25.618±0.265, and 0.681±0.007 (star 3), respectively at a concentration of 100 ug/mL. The most significant skin-lightening activity was observed in the ethanol extract Vc (Vc-ET), with an IC50 value inhibiting tyrosinase of 39.059 ug/mL. Regarding the anti-aging activity of the extracts, as determined by their antioxidant activity and inhibition of collagenase, elastase, and hyaluronidase, the most promising extracts were obtained from ethanol extract Va (Va-ET) and ethyl acetate extract Va (Va-EA), with IC50 values of 79.734, 76.838, 143.384, and 122.467 ug/ mL, respectively. The Va-ET was formulated into a gel dosage form, and there was a significant effect of the gelling agent on the SPF value of the extract after formulation (p < 0.05). All extracts exhibit activity as sunscreen, skin lightening, and anti-aging, with the Va-ET showing the highest efficacy. Gelling agents significantly influence the SPF value of extracts after formulation into the gel dosage form. Among them, F3, formulated from the Va-ET of P. scutellaroides using Carbopol-940 as a gelling agent, exhibits the highest efficacy.

Anti-wrinkle and Antioxidant Effects of Elderflower (Sambucus nigra) Water Extract as a Functional Ingredient of Cosmetics

Purpose: This study examined the potential of elderflower (<i>Sambucus nigra</i>) water extract (SNE) as a cosmetic material by evaluating its anti-wrinkle, antioxidant, and cytotoxic effects.Methods: We determined the antioxidant effect of SNE by measuring its 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical–scavenging and superoxide dismutase (SOD)-like activity and determined the anti-wrinkle effect of SNE by measuring its elastase inhibition activity. In addition, we assayed the viability of B16-F10 mouse melanoma and CCD-986sk human dermal fibroblast cells in the presence of SNE.Results: SNE showed a DPPH radical–scavenging activity of 77%, 76.4%, and 75.2% at a concentration of 1000, 500, and 250 μg/mL, respectively, and SOD-like activity of 78%, 70%, and 66% at a concentration of 200, 100, and 50 μg/mL, respectively. In addition, 500 μg/mL of SNE exhibited 48% elastase inhibition activity. Furthermore, B16-F10 cells showed a viability of 118%, 115%, and 109% when treated with 200, 100, and 50 μg/mL of SNE, respectively, and CCD-986sk cells showed high cell viability when treated with 200, 100, and 50 μg/mL of SNE.Conclusion: The results of this study indicate that SNE has excellent antioxidant and anti-wrinkle effects, confirming its high applicability as a functional cosmetic material.

The clinical effectiveness of sivelestat in treating sepsis patients with both acute respiratory distress syndrome and septic cardiomyopathy

BackgroundWe aimed to assess the efficacy of the neutrophil elastase inhibitor, sivelestat, in the treatment of sepsis-induced acute respiratory distress syndrome (ARDS) and septic cardiomyopathy (SCM).MethodsBetween January 2019 and December 2021, we conducted a randomized trial on patients who had been diagnosed with sepsis-induced acute respiratory distress syndrome (ARDS) and septic cardiomyopathy (SCM) at Wuhan Union Hospital. The patients were divided into two groups by random envelop method, the Sivelestat group and the Control group. We measured the serum concentrations of Interleukin (IL)-6, IL-8, Tumor necrosis factor-α (TNF-α), and High-mobility group box 1 (HMGB1) at five time points, which were the baseline, 12 h, 24 h, 48 h, and 72 h after admission to the ICU. We evaluated the cardiac function by sonography and the heart rate variability (HRV) with 24-hour Holter recording between the time of admission to the intensive care unit (ICU) and 72 h after Sivelestat treatment.ResultsFrom January 2019 to December 2021, a total of 70 patients were included in this study. The levels of IL-6, IL-8, and TNF-α were significantly lower in the Sivelestat group at different time points (12 h, 24 h, 48 h, and 72 h). HMGB1 levels were significantly lower at 72 h after Sivelestat treatment (19.46 ± 2.63pg/mL vs. 21.20 ± 2.03pg/mL, P = 0.003). The stroke volume (SV), tricuspid annular plane systolic excursion (TAPSE), early to late diastolic transmitral flow velocity (E/A), early (e’) and late (a’) diastoles were significantly low in the Control group compared with the Sivelestat group. Tei index was high in the Control group compared with the Sivelestat group (0.60 ± 0.08 vs. 0.56 ± 0.07, P = 0.029). The result of HRV showed significant differences in standard deviation of normal-to-normal intervals (SDNN), low frequency (LF), and LF/HF (high frequency) between the two groups.ConclusionsSivelestat can significantly reduce the levels of serum inflammatory factors, improve cardiac function, and reduce heart rate variability in patients with Sepsis-induced ARDS and SCM.

System proteinase/inhibitors in the pathogenesis of experimental kidney tuberculosis

BACKGROUND: Damage to the kidney parenchyma of various origins is associated with the accumulation of extracellular matrix proteins, the regulation of which involves the matrix metalloproteinase/inhibitor system. AIM: To evaluate the pathogenetic role of the proteinase/blood inhibitor system in experimental renal tuberculosis. MATERIAL AND METHODS: When modeling the process, a clinical strain 5582 of the Beijing genotype with multidrug resistance of mycobacteria was used. The study was conducted on 20 rabbits, divided into three groups: the first (n=6) included infected untreated animals; in the second (n=7) and third (n=7) anti-tuberculosis therapy was administered orally for 18.5 weeks; in the third, treatment was carried out in combination with a single injection of autologous mesenchymal stem cells into the ear vein. Blood levels of matrix metalloproteinases-1 and -9, tissue inhibitor of metalloproteinases-1, cystatin C and neutrophil elastase were determined. Morphological changes in the kidney parenchyma at the end of the experiment were assessed using 26 indicators, including tubulointerstitial and vascular changes. The median and interquartile ranges were calculated, the Kruskal–Wallis test, analysis of covariance, and the projective classification method were used. RESULTS: The progression of kidney tuberculosis was accompanied by a significant increase in the level of neutrophil elastase by 1.9 times, matrix metalloproteinase-9 by 2.2 times, and cystatin C by 1.5 times compared to the initial level. The treatment carried out in the second and third groups contributed to the normalization of these indicators. The concentrations of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in all three groups did not change relative to the baseline level. Covariance analysis showed that in the pathogenesis of renal tuberculosis changes in matrix metalloproteinase-1 (p=0.003), matrix metalloproteinase-9 (p=0.002), tissue inhibitor of metalloproteinases-1 (p=0.01) and cystatin C (p=0.0001) were primarily associated with vascular changes, found in all study groups. A linear combination containing three morphological characteristics allowed us to identify differences between the three groups with 90% accuracy. CONCLUSION: Renal tuberculosis, caused by a multidrug-resistant pathogen, is characterized by an imbalance in the proteinase/inhibitor system and impaired renal function, the degree of which is associated with the morphological characteristics of vascular and tubulointerstitial changes.

Skin anti-aging potential of Launaea procumbens extract: Antioxidant and enzyme inhibition activities supported by ADMET and molecular docking studies

Aging is a natural process that occurs in all living organisms. Particularly, the skin embodies aging since it serves as a barrier between the body and its surroundings. Previously, we reported the wound healing effect of Launaea procumbens and identified compounds therein. The study aims to explore the skin anti-aging properties of the plant extract. To that effect, the antioxidant potential of L. procumbens methanolic extract (LPM) was assessed using two complementary DPPH and FRAP assays. The enzyme inhibitory effect of the extract on collagenase, elastase, hyaluronidase, and tyrosinase was evaluated to assess the direct skin anti-aging effects. Similarly, the anti-inflammatory activity was evaluated to explore the indirect anti-aging effects via the assessment of extract inhibitory effects on cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX). In addition, ADMET and molecular docking studies were performed to explore the interaction mechanisms of identified compounds in LPM with target enzymes. LPM demonstrated significant antioxidant activity in DPPH (IC50 = 29.08 µg/mL) and FRAP (1214.67 µM FeSO4/g extract) assays. Plant extract showed significant inhibition of collagenase, elastase, hyaluronidase, and tyrosinase (IC50 = 52.68, 43.76, 31.031, and 37.13 µg/mL, respectively). The extract demonstrated significant COX-2 and 5-LOX inhibition capacity with IC50 values of 8.635 and 10.851 µg/mL, respectively. The molecular docking study revealed the high potential of the identified compounds to bind to the active sites of enzymes crucially involved in the skin aging process. ADMET analysis of the compounds revealed their good absorption, distribution, and metabolism profiles, and they were found to be safe as well. Study findings suggest L. procumbens as a promising source for the development of natural skin anti-aging and antioxidant compounds. This, in turn, may facilitate its incorporation into cosmetic formulations after further investigation.

Evaluation of potential anti-aging effects of Achillea phrygia Boiss. & Balansa (Asteraceae)

This study aims to determine the anti-aging effects of Achillea phrygia, an endemic plant, by evaluating its sun protection factor (SPF) level, antioxidant activity, total phenolic content, extracellular matrix-degrading enzymes (ECM) inhibition, genotoxic/anti-genotoxic, and cytotoxic activities. The SPF level was assessed using an in vitro quantitative method, while antioxidant capacity was determined through DPPH, β-carotene, and hydroxyl-radical (H2O2) scavenging assays. The total phenolic content was quantitatively conducted using the Folin Ciocalteu reagent. The inhibition of ECM-degrading enzymes was determined using matrix metalloproteinase-1 (MMP-1), hyaluronidase, and elastase enzymes. Genotoxic/anti-genotoxic properties were assessed using the AMES Salmonella/microsome assay, and cytotoxicity effects were assessed through the MTT assay. The results indicated that A. phrygia showed moderate SPF activity (SPF = 4.013) and exhibited IC50 values of 0.183 ± 0.03, 0.079 ± 0.51, and 1.18 ± 0.35 mg/mL for DPPH, β-carotene, and hydroxyl-radicals, respectively. The total phenolic content was measured to be 23.56 ± 1.42 mg GAE/g dry extract. Furthermore, the extract demonstrated inhibition of MMP-1 (47.98%) and elastase (39.2%) activities. Importantly, it did not induce DNA damage and showed antigenotoxic activity ranging from 10% to 65.6%. The cytotoxicity assay revealed an IC50 value of 42.41±4.05 µg/mL. These findings suggest that A. phrygia could be utilized as a cosmetic ingredient in skincare products due to its ability to protect against UV radiation, exhibit antioxidant properties, prevent extracellular matrix degradation, and inhibit DNA damage.

Ginseng fruit rare saponins (GFRS), a promising anti-wrinkle agent: Evidence of its antioxidant effect and its capacity to prevent matrix metalloproteinase (MMPs) expression in vitro and in vivo

Rare saponins show numerous significant biological activities, however, there has been no research on using ginseng fruit rare saponins (GFRS) to minimize skin wrinkles. This paper studied the anti-wrinkle effect, related mechanism, material basis of efficacy, and structure-activity relationship of GFRS in vivo and in vitro. The findings demonstrate that GFRS has potent collagenase and elastase inhibition capabilities in addition to superior ultraviolet absorption properties. Secondly, GFRS can strengthen the endogenous antioxidant defense system, thus lessening the UVB-induced ultraviolet damage that L929 cells and zebrafish experience. Matrix metalloproteinases (MMPs) studies have shown that GFRS could significantly inhibit the UVB-induced MMP1 and MMP9 proteins production. Finally, the main ginsenoside component in GFRS was identified by LC-MS, and 16 kinds of ginsenoside were analyzed, including rare ginsenoside such as Rg6, F4, and R/S-Rg3, which may be caused by the transformation of prototype saponins Re and Rd caused by citric acid heat treatment (CAHT). After doing a quantitative HPLC analysis on twelve of them, it was discovered that CAHT could enrich the content of rare saponins in GFS. By evaluating these ginsenoside’s anti-wrinkle properties. It is found that the main material basis for GFRS’s anti-wrinkle action may be ginsenoside Rg6 or F4. In summary, GFRS exhibits promising properties as an anti-wrinkle treatment because of its robust anti-oxidation, anti-collagenase, anti-elastase activity, favorable ultraviolet absorption effect, and capacity to regulate MMPs in vivo and in vitro models.

Valorization of Wine Lees: Assessment of Antioxidant, Antimicrobial and Enzyme Inhibitory Activity of Wine Lees Extract and Incorporation in Chitosan Films

Wine lees, as one of the main by-products of winery wastes, is considered a rich source of bioactive compounds such as polyphenols. Recovery of such bioactive compounds would amplify the valorization of winery by-products, as well as promote their use in cosmetics and health applications. Towards this aim, the objective of this work is the recovery of phenolic compounds from wine lees by ultrasound-assisted extraction. The effect of extraction time, and ethanol content on the extraction yield of total phenolics was investigated, while the phenolic profile of the extract was monitored by GC-MS. Under the optimized conditions, the total phenolic content was calculated at 302.6 mg gallic acid equivalents g− 1 dry extract; the dry extract was investigated for its biological activities, exhibiting excellent antioxidant and antimicrobial activity. Moreover, the extract was found to be a potent inhibitor of tyrosinase, elastase, collagenase, α-glucosidase, and α-amylase with IC50 values of 53.3, 83.5, 82.7, 19.0, and 31.3 µg mL− 1, respectively. Finally, the wine lees dry extract was incorporated into chitosan films, reinforcing the properties of the films, and providing them with excellent antioxidant activity. The results support that wine lees extract exhibits high antioxidant activity and can be used as a highly efficient inhibitor of enzymes with pharmaceutical and health interest. Moreover, the chitosan film enriched with wine lees extract is a promising material with high antioxidant potential and can lead to the development of novel food packaging films in the food industry.Graphical

Enhancing the Antioxidant, Anti-tyrosinase and Anti-elastase Activity of Pigmented Rice Extracts by Sonication, Heating, Enzyme Digestion and Fermentation

In the modern cosmetics industry, there exists a major trend for replacement of synthetic chemicals with plant-based ingredients in skincare formulation. This research aimed to assess the efficiency of different methods to extract bioactive compounds (as measured by total phenolic content; TPC and total flavonoid content; TFC) responsible for three important cosmeceutical properties (antioxidation, anti-tyrosinase, and anti-elastase) from selected rice grains. The grains of two varieties of purplish-black rice (Neaw Leum Pua; NLP) and black rice (Hom Nil; HN) were extracted with phosphate buffered saline (PBS) and then supplementary treated with sonication, heating, and proteolytic enzyme digestion or fermented with Aspergillus niger. The results showed that sonication slightly enhanced TPC and TFC, but significantly increased antioxidant capacity and anti-tyrosinase activity. The high temperature of 60℃ and 100℃ significantly increased antioxidant capacity based on FRAP assay but reduced that based on DPPH radical scavenging assay. Anti-tyrosinase activity was enhanced by high temperature but that of anti-elastase was inhibited. The addition of papain to the seed extracts slightly enhanced anti-elastase and anti-tyrosinase activity. Notably, fermented rice grains exhibited much greater yields of TPC and TFC, and higher biochemical activities than other methods. The fermented rice exhibited the highest TFC after 3 days; the highest TPC, antioxidant capacity, and tyrosinase inhibition after 6 days; and the highest elastase inhibition activity after 9 days. Therefore, fermentation was the most promising technique to enhance all three desirable properties for cosmetic ingredients when pigmented rice was used as the raw material.

Inhibition of neutrophil elastase by the bacterial serine protease inhibitor ecotin in cystic fibrosis airway samples

Abstract Cystic fibrosis (CF), a prevalent fatal genetic condition in North America, disrupts respiratory function by impeding chloride ion transport across mucosal surfaces. This leads to the thickening of mucus, resulting in constant lung inflammation and elevated levels of neutrophils. Neutrophil elastase (NE), a potent serine protease crucial for host defense, is released by these neutrophils. However, in excess, NE can cause detrimental host tissue damage. Abundant in CF airways, NE is a key predictor of lung disease progression. While past studies have explored targeting NE in CF, no optimal inhibitors have reached clinics. Our study tested ecotin, a bacterial periplasmic serine protease inhibitor, for its potential to impede NE activity in CF airway samples and curb NE release from human neutrophils. Results show that ecotin significantly reduces NE activity in sputum samples obtained from several CF patients. Additionally, we found that ecotin inhibits NE release from neutrophils exposed to CF lung stimuli like Staphylococcus aureus bacteria. These results establish ecotin as a promising NE inhibitor in CF with clinical potential.

The beneficial effects of neutrophil elastase inhibitor on gastrointestinal dysfunction in sepsis.

To investigate the effects of neutrophil elastase inhibitor (sivelestat sodium) on gastrointestinal function in sepsis. A reanalysis of the data from previous clinical trials conducted at our center was performed. Septic patients were divided into either the sivelestat group or the non-sivelestat group. The gastrointestinal dysfunction score (GIDS), feeding intolerance (FI) incidence, serum levels of intestinal barrier function and inflammatory biomarkers were recorded. The clinical severity and outcome variables were also documented. A total of 163 septic patients were included. The proportion of patients with GIDS ≥2 in the sivelestat group was reduced relative to that in the non-sivelestat group (9.6% vs. 22.5%, p = 0.047) on the 7th day of intensive care unit (ICU) admission. The FI incidence was also remarkably reduced in the sivelestat group in contrast to that in the non-sivelestat group (21.2% vs. 37.8%, p = 0.034). Furthermore, the sivelestat group had fewer days of FI [4 (3, 4) vs. 5 (4-6), p = 0.008]. The serum levels of d-lactate (p = 0.033), intestinal fatty acid-binding protein (p = 0.005), interleukin-6 (p = 0.001), white blood cells (p = 0.007), C-reactive protein (p = 0.001), and procalcitonin (p < 0.001) of the sivelestat group were lower than those of the non-sivelestat group. The sivelestat group also demonstrated longer ICU-free days [18 (0-22) vs. 13 (0-17), p = 0.004] and ventilator-free days [22 (1-24) vs. 16 (1-19), p = 0.002] compared with the non-sivelestat group. In conclusion, sivelestat sodium administration appears to improve gastrointestinal dysfunction, mitigate dysregulated inflammation, and reduce disease severity in septic patients.

Unveiling the metabolites underlying the skin anti-ageing properties of Cytinus hypocistis (L.) L. through a biochemometric approach

The genus Cytinus, recognised as one of the most enigmatic in the plant kingdom, has garnered attention for its bioactive potential, particularly its skin anti-ageing properties. Despite this recognition, much remains to be accomplished regarding deciphering and isolating its most active compounds. This study aimed to identify the compounds responsible for C. hypocistis skin anti-ageing potential. Using multivariate analysis, a biochemometric approach was applied to identify the discriminant metabolites by integrating extracts' chemical profile (Liquid Chromatography-High-Resolution Mass Spectrometry, LCHRMS) and bioactive properties. The identified bioactive metabolite was structurally elucidated by 1D and 2D Nuclear Magnetic Resonance (NMR). Among the studied bioactivities, the anti-elastase results exhibited a significant variation among the samples from different years. After the biochemometric analysis, the compound 2,3:4,6-bis(hexahydroxydiphenoyl)glucose, with a molecular mass of 784.075 Da, was structurally elucidated as the discriminant feature responsible for the outstanding human neutrophil elastase inhibition. Remarkably, the subfraction containing this compound exhibited a tenfold improvement in neutrophil elastase inhibition efficacy compared to the crude extract; its effectiveness fell within the same range as SPCK, a potent irreversible neutrophil elastase inhibitor. Moreover, this subfraction displayed no cytotoxicity or phototoxicity and excellent efficacy for the tested anti-ageing properties. Hydrolysable tannins were confirmed as the metabolites behind C. hypocistis skin anti-ageing properties, effectively mitigating critical molecular mechanisms that influence the phenotypically distinct ageing clinical manifestations. Pedunculagin was particularly effective in inhibiting neutrophil elastase, considered one of the most destructive enzymes in skin ageing.

Nanoemulsions Containing Passiflora quadrangularis L. Fruit Extracts for Cosmetic Application and Skin Efficacy Study

This study aimed to determine the anti-collagenase, anti-elastase, and anti-hyaluronidase activities of Passiflora quadrangularis fruit extracts (epicarp, mesocarp, endocarp, and seed), develop stable nanoemulsions, and evaluate the efficacy of the nanoemulsions containing extracts in human volunteers. The results indicated that the epicarp and seed extracts exhibited collagenase, elastase, and hyaluronidase inhibition effects. Gallic acid was identified in the extracts, with the highest concentration found in the endocarp extract (1449.35 mg gallic acid/g extract) followed by the seed extract (839.63 mg gallic acid/g extract). The endocarp and seed extracts demonstrated good stability at different temperatures. Consequently, these extracts were selected for incorporation into nanoemulsions due to their high yield, excellent anti-tyrosinase and anti-aging activity, and good stability, making them suitable for cosmetic products. The nanoemulsions were prepared using the ultrasonication method, resulting in a particle size of 133.1 ± 0.8 nm, a polydispersity index of 0.33 ± 0.03, and a zeta potential of −61.8 ± 0.6 mV, indicating good stability. The nano-serum containing extracts was non-irritating and safe for the skin. Skin evaluation among human volunteers after 60 days of application revealed improvements in skin hydration and reduction in wrinkles. In conclusion, nanoemulsions containing P. quadrangularis fruit extracts have the potential to be effective cosmetic products.

A neutrophil elastase inhibitor, sivelestat, attenuates sepsis-induced acute kidney injury by inhibiting oxidative stress

BackgroundSivelestat, a selective inhibitor of neutrophil elastase (NE), can mitigate sepsis-related acute lung injury. However, the role of sivelestat in inhibiting oxidative stress and attenuating sepsis-related acute kidney injury (AKI) remains unclear. Here, we reported the effects of sivelestat against oxidative stress-induced AKI by suppressing the production of oxidative stress indicators. Materials and methodsA male Sprague-Dawley rat model of sepsis was established by cecal ligation and puncture (CLP). Sivelestat or normal saline was administered into jugular vein with a sustained-release drug delivery system. Indicators of inflammation and AKI, including white blood cells (WBC), neutrophils, lymphocytes, C-reactive proteins (CRP), procalcitonin (PCT), blood urea nitrogen (BUN), creatinine (Cr) and uric acid (UA), were assessed at 24 h post-sivelestat treatment. Indicators of liver injury, including direct bilirubin (DBIL), indirect bilirubin (IBIL), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), were also assessed at 24 h post-sivelestat treatment. Indicators of oxidative stress, including superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px), were assessed at 12 h and 24 h post-sivelestat treatment. At 24 h post-sivelestat treatment, H&E staining of kidney and liver tissue was performed to observe pathological alterations. ResultsAt 24 h post normal saline or sivelestat (0.2 g/kg body weight) treatment, WBC, neutrophil, CRP, PCT, MDA, BUN, Cr, UA, AST, ALT, DBIL and IBIL were increased, while SOD and GSH-Px were decreased, in septic rats treated with normal saline compared with that in non-septic rats treated with normal saline (all p < 0.05). The changes of these indicators were reversed in septic rats treated with sivelestat compared with that in septic rats treated with normal saline (all p < 0.05). Similar results were found regarding the levels of oxidative stress indicators at 12 h post-sivelestat treatment. The degenerative histopathological changes in both kidney and liver tissues were ameliorated upon sivelestat treatment. ConclusionsSivelestat plays a protective role in sepsis-related AKI by inhibiting oxidative stress. Our study reveals a possible therapeutic potential of sivelestat for oxidative stress-induced AKI.

Features of the spectrum of immune markers in patients with juvenile depression with clinically high risk of psychosis

IntroductionIdentification of biomarkers associated with the risk of psychosis manifestation in juvenile patients with depression may contribute to a better understanding of the pathogenesis of mental disorders and early diagnosis.ObjectivesTo determine the level of pro-inflammatory and anti-inflammatory cytokines and other inflammatory indicators in the plasma of juvenile patients with depression and clinically high risk of psychosis, and to study the correlation of these markers with the severity of psychopathologic symptoms.Methods80 young men aged 16-24 years with the first depressive episode (F32.1-2, F32.38, F32.8) were examined. Based on the severity of attenuated psychotic symptoms (APS) in the structure of depression according to the SOPS scale, all patients were divided into two groups - with clinically high risk of psychosis (n=58) and with depression without APS (n=22). The HDRS-21 and SANS scales were also used for psychometric assessment. Serum level of cytokines TNF-α, IL-6, IL-8, IL-10, TNF-α/IL-6 ratio, TNF-α/IL-10 ratio, leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) activity, C-reactive protein (CRP) concentration, and the level of autoantibodies to S-100B protein were determined.ResultsBoth groups of patients showed a high level of inflammation assessed by LE and α1-PI activity (p&gt;0.05). Significantly higher level of IL-6 (p=0.03), CRP concentration (p=0.026) and TNF-α/IL-10 ratio (p=0.032) were found in patients with clinically high risk of psychosis. This group was also characterised by high level of autoantibodies to the S-100B protein compared to patients with depression without APS (p=0.048). In the high clinical risk group, correlations were found between the SOPS positive subscale score and the level of TNF-α (R=0.32, p=0.017), IL-8 (R=-0.3, p=0.034), TNF-α/IL-6 ratio (R=0.30, p=0.021) and TNF-α/IL-10 ratio (R=0.32, p=0.014). The SOPS negative subscale score correlated with CRP concentration (R=0.3, p=0.043). The SOPS total score correlated with TNF-α/IL-10 ratio (R=0.31, p=0.021). In this group of patients, the level of IL-10 was found to correlate with the duration of the disease (R=0.48, p&lt;0.001). In patients with depression without APS, the level of IL-6 was correlated with the severity of depression according to the HDRS scale, and the level of TNF-α was associated with the duration of the depressive episode (R=0.51, p=0.029).ConclusionsThe obtained results confirm the involvement of inflammation in the development of juvenile depression. Qualitative and quantitative characteristics of the spectrum of immune markers and the cytokine profile, and correlations with the severity of psychopathologic symptoms were revealed in patients with clinically high risk of psychosis.Disclosure of InterestNone Declared

Subgroups of patients with late onset schizophrenia-like psychoses revealed by the analysis of glutathione-dependent enzymes and inflammation markers

IntroductionWhile chronic inflammation and enhanced imbalance of pro- and antioxidant, including glutathione-dependent, systems contribute substantially to pathogenesis of mental disorders in old age, extent of oxidative stress and degree of inflammatory processes severity are varying among patients with late onset schizophrenia.ObjectivesRevealing various phenotypes in patients with late onset schizophrenia basing on measurement of activity levels for blood glutathione-dependent enzymes and inflammation markers and analysis of their links with clinical features of the patients.MethodsOf 59 studied women patients 34 were with late onset (after 40 years) and 25 with very late onset (after 60 years) schizophrenia or schizophrenia-like psychoses (F20; F22.8; F25; F23; F06.2 by ICD-10). 34 mentally healthy women elder than 50 years comprised controls. Glutathione reductase (GR), glutathione-S-transferase (GST), neutrophil elastase (NE), and α1-roteinase inhibitor (α1-PI) activities were measured in blood. PANSS, CDSS and CGI-S were used to assess the severity of psychotic symptoms, depression and treatment effectiveness.ResultsIn the whole group of patients, GR was lower (p&lt;0.05), and α1-PI was higher (p&lt;0.0001) than in control group. Clustering the patients by their biochemical and immunological signs revealed two clusters (C1, n=34, and C2, n=25) significantly differing by GST (p&lt;0.0001), NE (p&lt;0.0001), and α1-PI (p&lt;0.001) activities. As compared with controls, GST and α1-PI were higher (p&lt;0.05 and p&lt;0.0001), and NE was lower (p&lt;0.05) in C1. As compared with controls, GR activity was lower (p&lt;0.05), NE activity was higher (p&lt;0.001), and α1-PI activity was much higher (p&lt;0.001) in C2. Patients of C1 and C2 did not differ in age, diagnosis, severity of the disease, but differed in clinical features of the course of the disease: significantly more patients with very late onset schizophrenia (76%) were met in C1 (χ2=13.41, p&lt;0.001). Also, different clinical-biological correlations were found in these clusters. Particularly, negative correlations of baseline NE activity with PANSS general psychopathology subscale scores (R=-0.39, p&lt;0.05) and with total PANSS scores (R=-0.39, p&lt;0.05) were found in C1. Positive correlation of GST activity with PANSS positive subscale score was found in C2 (R=0.43, p&lt;0.05).ConclusionsThe revealed clusters differ in the extent of the glutathione antioxidant system impairment and in levels of the immune response markers. The revealing of the patient subgroups on the basis of biological markers reflecting impairments in metabolic and immune systems can represent interest in the search for individual treatment approaches.Disclosure of InterestNone Declared

Immunological predictors of rhythmic transcranial magnetic stimulation (rTMS) efficiency in patients with treatment-resistant schizophrenia

IntroductionModel and clinical studies demonstrate the efficiency of rhythmic transcranial magnetic stimulation (rTMS) in diseases associated with neuroinflammation. The therapeutic potential of rTMS is related to modulation of neuroplasticity in the CNS, activation of neurogenesis and reduction of neuroinflammatory processes. Presumably, one of the factors that determines the efficiency of rTMS can be the features of the immune status of patients.ObjectivesTo reveal the features of the spectrum of inflammatory markers in patients with treatment-resistant schizophrenia with different efficiency of rTMS.Methods 31 male patients aged 16 to 47 years (mean age 29.9 ± 8.4 years) with treatment-resistant schizophrenia who developed a first psychotic episode in adolescence (19-25 years) were examined. The course of rTMS was conducted for 3 weeks (15 sessions). Depending on the dynamics of clinical and psychometric parameters after the course of rTMS, the patients were divided into three groups: group 1 - with worsening of clinical condition (n=8); group 2 - without therapeutic effect (n=12); group 3 - with good therapeutic response (n=11). Before rTMS, leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) activity, and the levels of autoantibodies to S-100B protein and myelin basic protein (MBP) in the plasma of patients were determined. The parameters of 18 healthy male donors without clinical signs of psychiatric and somatic pathology were used as controls.ResultsAll groups of patients were characterised by moderate and high levels of immune system activation, determined by a complex of inflammatory and autoimmune markers. At the same time, the high level of immune system activation in patients with low MTR efficiency was associated with low LE activity in plasma (within the reference range or below the lower limit - 200.6 (168.5- 220.3) nmol/min·mL), which was not consistent with the overall level of inflammation. This group of patients also showed high levels of antibodies to MBP compared to control values (p&lt;0.05). The low LE activity can be explained by the transmigration of neutrophils from the blood to the brain due to a critical increase in the permeability of the blood-brain barrier, which is largely controlled by LE.ConclusionsThe study confirmed the participation of immune mechanisms in the formation of therapeutic resistance in schizophrenia and revealed the characteristics of the spectrum of immune markers in patients with low efficiency of rTMS.Disclosure of InterestNone Declared

Antioxidant and in vitro cosmeceutical activities of germinated oats

AbstractBackground and ObjectivesOat (Avena sativa L.) is well known for its biofunctional properties, primarily attributed to cereal phytochemicals. Germination may amplify the formation of secondary metabolites in oat, enhancing its nutritional and bioactive quality. Despite this, the impact of germination of oat on skin health remains unknown. This study investigated how germination influences the in vitro antioxidant activity and enzyme inhibitory activity related to skin health (tyrosinase and elastase).FindingsOat was germinated at 25°C for 6 days, and the germinated samples were collected every 2 days. The samples were then divided into kernel and radicle (including germ) fractions for analysis. The radicle fraction exhibited significantly higher antioxidant activity (2,2‐diphenyl‐1‐picrylhydrazyl [DPPH] radical scavenging) and in vitro cosmeceutical activity compared to the whole kernel. Phenolic acids were identified to be responsible for the antioxidant activity and tyrosinase inhibition at all germination times, whereas phytosterols including β‐sitosterol played the key role in elastase inhibition. Avenanthramides, synthesized during oat germination, were proved to be crucial in inhibiting the enzymes deteriorating skin health.ConclusionsThis study revealed that germination, as a bioconversion process, enhanced antioxidant activity and cosmeceutical activity of oat by inhibiting the key enzymes responsible for skin aging. However, further research is imperative to fully comprehend the roles and mechanisms of these substances produced during germination.Significance and NoveltyThis study revealed the impacts of germination with cereals, offering valuable insights into the strategies for future investigations in the field of skin health and bioactive cereal compounds.

Sivelestat (Neutrophil Elastase Inhibitor) as an Anti-inflammatory and Anti-viral Agent: An In Silico Study.

Background Sivelestat is a potent and specific neutrophil elastase inhibitor. It is clinically used in treating lung injury and respiratory distress syndrome. This engaged us to undertake the present study in which sivelestat was studied as an anti-inflammatory and anti-viral agent. Methodology The docking study of sivelestat on matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), chikungunya virus nonstructural protein-2 (CVnsP2) protease, and influenza A (H1N9) virus neuraminidase was assessed using the Chemistry at Harvard Macromolecular Mechanics (CHARMM) Dock (CDOCK) method. Furthermore, molecular physicochemical; bioactivity; absorption, distribution, metabolism, and excretion (ADME); toxicity; and Search Tool for Interacting Chemicals (STITCH) analyses were performed by using the Molinspiration (Molinspiration Cheminformatics, Slovensky Grob, Slovak Republic), SwissADME SwissADME (Swiss Institute of Bioinformatics, Quartier Sorge - Bâtiment Amphipôle, Switzerland), pkCSM (University of Melbourne, Melbourne, Australia), and STITCH-free online tools. Results The molecular physicochemical assessment of the ligand (sivelestat) showed no (zero) violation and agreed with the thumb rule of five, otherwise known as Lipinski's rule of five. ADME prediction of the ligand (sivelestat) is shown to possess a low gastrointestinal absorption (GIA) property. Similarly, toxicity analysis of the ligand (sivelestat) is predicted to have a hepatotoxicity effect. STITCH analysis reveals that the ligand (sivelestat) has exhibited interactions with the three human proteins. Conclusions The present molecular docking studies showed that the ligand (sivelestat) has successfully docked with all four enzymes of interest. Hence, the current finding has provided a good understanding of sivelestat as an effective suppressor activity against all four enzymes: MMP-2, MMP-9, CVnsP2 protease, and influenza neuraminidase.