Cyclooxygenase-2 Activity Research Articles

Biogenic Selenium Nanoparticles Synthesized by Lactobacillus casei ATCC 393 Alleviate Acute Hypobaric Hypoxia-Induced Intestinal Barrier Dysfunction in C57BL/6 Mice.

Exposure to hypobaric hypoxia at high altitude will cause different tissue and organ damage over a long period of time. Studies have shown that hypobaric hypoxia can cause severe primary intestinal barrier dysfunction, and then cause multiple organ dysfunction. Our previous research showed that selenium nanoparticles (SeNPs) synthesized by Lactobacillus casei ATCC 393 (L. casei ATCC 393) can effectively alleviate intestinal barrier dysfunction caused by oxidative stress and inflammationin mice. This study was conducted to investigate the protective effect of biological SeNPs synthesized by L. casei ATCC 393 on intestinal barrier function in acute hypobaric hypoxic stress mice. The results showed that compared with the hypobaric hypoxic, the SeNPs synthesized by L. casei ATCC 393 by oral administration could effectively alleviate the shortening of intestinal villi, which decreased the level of diamine oxidase (DAO) and myeloperoxidase (MPO), and the expression level of tight junction protein in ileum was increased. In addition, SeNPs significantly increased the activities of superoxide dismutase (SOD), cyclooxygenase (COX-1) and glutathione peroxidase (GPx), and decreased the level of malondialdehyde (MDA), and inhibit the increase of hypoxia related factor. SeNPs effectively regulate the intestinal microecology disorder caused by hypobaric hypoxia stress, and maintain the intestinal microecology balance. In addition, oral administration of SeNPs had better protective effect on intestinal barrier function of mice under hypobaric hypoxia stress. These results suggested that SeNPs synthesized by L. casei ATCC 393 can effectively alleviate the damage of intestinal barrier function under acute hypobaric hypoxic stress, which may be closely related to the antioxidant activity of SeNPs.

Chemical profiling, in vitro biological activities and Pearson correlation between chemical profiling and anticancer activities of four Abies species from Turkey

Abies species are known to have various bioactive compounds and pharmacological activities. In the manuscript, their bioactive components and anticancer activities of four unexamined Abies in Anatolia ( Abies cilicica subsp. isaurica, Abies nordmanniana subsp. nordmanniana, Abies nordmanniana subsp. bornmuelleriana, Abies nordmanniana subsp. equi-trojani ) were investigated. The principal component analysis and drug likeness properties of the specific bioactive compounds were conducted on our datas. The results showed that solvent polarity affected the phytochemical composition of extracts and thus their anticancer activity. Gentisic acid, catechin, palmitic acid, stearic acid, oleic acid, caryophyllene, caryophyllene oxide and eicosene were identified as major components. The principal component analysis of the specific bioactive compounds results indicated that the compounds are able to have an effective anticancer activity against HeLa an C6 cells. At the same time, the drug likeness properties results showed that these compounds have a potential drug properties. Also, docking result demonstrated that oleic acid, stearic acid, catechin, palmitic acid and eicosene can perform a strong synergistic effect against Cyclooxygenase-2 activity. Hovewer, oleic acid had the best binding poses against six targets. According to the findings, this four Abies species could be considered for further phytopharmaceutical researchs. • The four Abies species from Turkey were researched. • GC–MS and HPLC-TOF/MS were used to determine phytochemicals of the extracts. • Abies ssp.'s anticancer effects against C6 and Hela cells were evaluated. • Molecular dockings showed interactions of anticancer effects and phytochemicals. • Abies ssp. could be considered for further pharmaceutical researchs.

A novel method for quality consistency evaluation of Glycyrrhiza formula granules based on a spectrum-effect study.

Quality consistency of Glycyrrhiza formula granules is essential for guaranteeing clinical efficacy. However, a suitable method to accurately and conveniently evaluate the consistency of the clinical efficacy of Glycyrrhiza formula granules is currently not available. This study established a method for the simultaneous determination of 12 active components in Glycyrrhiza formula granules using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry. The rate of inhibition of cyclooxygenase-2 by different batches of Glycyrrhiza formula granules was determined. Near-infrared spectra were collected for different batches of Glycyrrhiza formula granules to detect their biological activity in the inhibition of cyclooxygenase-2. The quality consistency of the 11 batches of Glycyrrhiza formula granules was evaluated using principal component and correlation analyses. The results showed significant differences in the formula granules of Glycyrrhiza uralensis produced by the different manufacturers. Some differences were also observed among batches of formula granules produced by the same manufacturer. Correlation analysis of the chemical components and cyclooxygenase-2 activity showed that glycyrrhizic acid, liquiritin, and isoliquiritin were the main active components of Glycyrrhiza. Correlation analysis of the near-infrared spectra and cyclooxygenase-2 inhibition activity showed a high correlation between the active components and three characteristic bands: 3383-3995, 4227-4651, and 5315-5878cm-1 . In this study, the main active anti-inflammatory components of Glycyrrhiza granules were screened. Thus, the near-infrared spectrum and characteristic active band of multi-index active components can be used to quickly detect the quality consistency of Glycyrrhiza formula granules, thereby improving the ability to control the quality and consistency of these granules.

Platelet-rich plasma counteracts interleukin-1 induced inhibition of collagen biosynthesis through recovery of impaired β1-integrin signaling and prolidase activity in human skin fibroblasts.

Although inflammation is the first step in wound healing, it contributes to down-regulation of collagen biosynthesis delaying the process of tissue regeneration. The study was conducted to evaluate the effects of platelet-rich plasma (PRP) on interleukin-1β (IL-1β) – dependent inhibition of collagen synthesis, prolidase activity, and β1-integrin signaling in cultured human skin fibroblasts. PRP was obtain using the SmartPReP®2 Autologous Platelet Concentrate+ System. Collagen biosynthesis and prolidase activity were measured in confluent human skin fibroblast cultures with IL-1β, PRP, hyaluronic acid (HA), and mixtures of IL-1β with PRP or HA. Immunofluorescence bioimaging analysis was employed to evaluate expression of β1 integrin receptor, cyclooxygenase-2 (COX-2) and nuclear factor-κB (NF-κB) protein. Incubation of fibroblasts with 2% PRP for 24 h contributed to about 500% increase in collagen biosynthesis and a significant increase in the expression of β1-integrin receptor and prolidase activity, compared to the control cells. In the cells treated with IL-1β, collagen biosynthesis, β1-integrin receptor expression, and prolidase activity were decreased while COX-2 and NF-κB expressions were significantly increased. All these processes were recovered to control values by PRP or HA; however, PRP was found to act more effectively than HA. It was found that PRP counteracted IL-1β -dependent inhibition of collagen synthesis through recovery of β1-integrin receptor expression and prolidase activity and down-regulation of COX-2 and NF-κB expressions in cultured fibroblasts. The data suggest that PRP evoke anti-inflammatory activity that is desirable in tissue regeneration.

New thiophene derivative augments the antitumor activity of γ-irradiation against colorectal cancer in mice via anti-inflammatory and pro-apoptotic pathways

BackgroundColorectal cancer (CRC) is one of the most common types of cancer worldwide and the second cause of cancer-related deaths. It usually starts as an inflammation that progresses to adenocarcinoma. The goal of the present study was to investigate the antitumor efficacy of a new thiophene derivative against CRC in mice and explore the possible associated molecular pathways. The potential of this thiophene derivative to sensitize the CRC tumor tissue to a low dose of gamma irradiation was also investigated.MethodsAdult male mice were divided into seven groups; control, group treated with dimethylhydrazine (DMH) for the induction of CRC. The DMH-group was further divided into six groups and treated with either cisplatin, thiophene derivative, γ-irradiation, cisplatin + γ-irradiation, thiophene derivative + γ-irradiation, or left untreated.ResultsDMH induced CRC as evidenced by the macroscopic examination of colon tissues and histopathology, and elevated the activities of cyclooxygenase2 (COX2) and nitric oxide synthase (iNOS). DMH also elevated kirsten rat sarcoma (KRAS) and downregulated the peroxisome proliferator activated receptor (PPARγ) as shown by RT-PCR and Western blotting. DMH exerted anti-apoptotic activity by reducing the expression of phosphorylated p53 and cleaved caspase3 at the gene and protein levels. The flow cytometry analysis showed that DMH elevated the necrosis and reduced the apoptosis compared to the other groups. The colon tissue from DMH-treated mice showed hyperplasia, aberrant crypt foci, loss of cell polarity, typical CRC of grade 4 with lymphocytes and macrophages infiltrating mucosa, muscularis mucosa, and submucosa score 3. Treatment with thiophene derivative or γ-irradiation ameliorated most of these deleterious effects of DMH. The concomitant action of thiophene derivative + γ-irradiation was typified by the better amelioration of tumor incidence and multiplicity, iNOS, PPARγ, p53, caspase 3, and histopathology of colon.ConclusionTaken together, the new thiophene derivative is a promising therapeutic candidate for treatment of colorectal cancer in mice. It also sensitizes the CRC tumor to the ionizing radiation through anti-inflammatory and pro-apoptotic pathways.

Betulin and Its Derivatives Reduce Inflammation and COX-2 Activity in Macrophages

Betulin is a heavily studied natural compound for its use as an anticancer or pro-regenerative agent. The structural similarity between betulin to steroids gives rise to the idea that the substance may as well act as an anti-inflammatory drug. This study is the first to describe the anti-inflammatory properties of betulinic acid, betulin, and its derivatives with amino acids 1,4-diaminebutane (Dab), 1,3-diaminepropane (Dap), Ornithine (Orn), and lysine (Lys) on murine macrophages from lymphoma site. The compounds were compared to dexamethasone. To establish the response of the macrophages to the natural compounds, we tested the viability as well as sensitivity to the inflammatory signaling (IFNγR). IL-6 secretory properties and HSP-70 content in the cells were examined. Furthermore, we characterized the effects of compounds on the inhibition of cyclooxygenase-2 (COX-2) activity both in the enzymatic assays and molecular docking studies. Then, the changes in COX-2 expression after betulin treatment were assessed. Betulin and betulinic acid are the low-cytotoxicity compounds with the highest potential to decrease inflammation via reduced IL-6 secretion. To some extent, they induce the reorganization of IFNγR with nearly no effect on COX-2 activity. Conversely, Bet-Orn and Bet-Lys are highly cytotoxic and induce the aggregation of IFNγR. Besides, Bet-Lys reduces the activity of COX-2 to a higher degree than dexamethasone. Bet-Orn is the only one to increase the HSP-70 content in the macrophages. In case of IL-6 reduction, all compounds were more potent than dexamethasone.

Diosmetin alleviates nonylphenol-induced liver damage by improving biochemical, inflammatory, apoptotic and histological profile in rats

Nonylphenol (NP) is an environmental pollutant that is recognized for its hazardous effects on humans and animals. NP has potential to induce oxidative stress that leads to hepatic toxicity. Diosmetin (DIOS) is a naturally occurring bioflavonoid that possesses several biological properties. The current study was designed to ascertain the curative effects of DIOS against NP prompted hepatotoxicity in rats. 32 male albino rats were randomly categorized in 4 groups i.e., control (0.1 % DMSO), NP (50 mg/kg), NP + DIOS (50 mg/kg + 100 mg/kg) and DIOS (100 mg/kg) group. Our results revealed that NP instigated substantial reduction in the antioxidant enzymes activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GSR), glutathione peroxidase (GPx), glutathione S-transferases (GST) and glutathione (GSH). NP administration raised the levels of reactive oxygen species (ROS) as well as the levels of malondialdehyde (MDA). Treatment with DIOS significantly (p < 0.05) recovered activities of antioxidant enzymes, ROS, and TBARS levels. Furthermore, DIOS treatment ameliorated the NP-induced increased level of inflammatory markers i.e., nuclear factor kappa B (NF-κB), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFα) and cyclooxygenase-2 (COX-2) activity. In addition, DIOS co-treatment also recovered the NP-provoked escalated levels of pro-apoptotic proteins (Bax, caspase-3 and caspase-9) and substantially reduced level of anti-apoptotic protein (Bcl-2) to normal levels. Besides, DIOS treatment recovered the potential histopathological damages in liver tissues. Therefore, DIOS might be an effective therapeutic agent for alleviating NP-induced hepatic toxicity due to its antioxidant, anti-inflammatory and anti-apoptotic potential.

Vitex trifolia as Cyclooxygenase-2 Inhibitors in Anti-Inflammatory Drug Discovery

Inflammation is involved in several stages of diseases such as cardiovascular diseases, asthma, diabetes, neurodegenerative diseases, or cancer. Cyclooxygenase-2 (COX-2) is an important enzyme that catalyzes the production of prostaglandins from arachidonic acid and has an important role in various inflammatory-related pathologies. The selective suppression of COX-2 over COX-1 is thought to be a useful approach for treating inflammation. Vitex trifolia is traditionally used for various inflammatory ailments, thus giving scope for anti-inflammatory studies. Since the action of Ethanol extract of Vitex trifolia (EEVT) on COX-2 has not been previously described. In this study, we further investigated the effect of Cyclooxygenase-2 inhibitory effects spectrophotometrically in ELISA readers and evaluated anti-inflammatory activity in the carrageenan-induced rat paw edema model. The edema volume was measured using a plethysmometer. The results showed that the EEVt (200, 400, and 800 mg/kg BW) had significant anti-inflammatory activity and the EEVt demonstrated a COX-2 Inhibitory effect. The percentage of COX-2 inhibition was 8,04%, 20,74%, 21,92%, and the percentage of inhibition of paw edema was 39,56%, 45,60%, and 40,56%, respectively. These findings suggest that Vitex trifolia leaves extracts to possess promising and remarkable anti-inflammatory activity, which is directed against the enzymatic activity of COX-2.

Interaction of meloxicam and phenylbutazone on the level of cyclooxygenase-2 in mice

The reason for the recent study was to inspect the therapeutic efficacy of meloxicam and phenylbutazone alone with their analgesic interaction and their subsequent inhibitory interaction at the level of cyclooxygenase-2 in mice. Meloxicam and phenylbutazone had the analgesic-median effective doses (ED50s) of 15.57 and 119.73 mg/kg, i.p., respectively, given once to mice separately as determined by the up-and-down procedure using a hot plate method. The estimated analgesic ED50s for meloxicam and phenylbutazone combination were at 12.84 and 98.75 mg/kg, i.p., correspondingly when given together at a ratio of 1:1 of their ED50s. The isobolographic analysis reveals that the analgesic interaction between meloxicam and phenylbutazone was antagonistic, as indicated by the interaction index of 1.65. The ELISA technique was used to estimate the cyclooxygenase-2 activity, reflecting that meloxicam or phenylbutazone significantly inhibited the cyclooxygenase-2 activity by 72 and 90%, respectively, compared to the control group. The combination composed of meloxicam and phenylbutazone has a lower limit of inhibition of the cyclooxygenase-2 activity (33%) in comparison to meloxicam or phenylbutazone. Meloxicam and phenylbutazone coadministration were significantly different from the control, meloxicam, and phenylbutazone groups concerning the cyclooxygenase-2 activity in mice. The sum of the data concluded that meloxicam and phenylbutazone have an excellent analgesic efficacy when administered alone. In contrast, the mixture of these two drugs has no benefit because of the antagonistic interaction at cyclooxygenase-2 in mice.

English

Aflatoxin B1 (AFB1) is a potent mycotoxin in humans and animals. The exposure to AFB1 is evidenced to implicate multi-organ toxicity in humans and animals, particularly hepatotoxicity. Genkwanin (GNK) is a bioactive non-glycosylated flavonoid with potential pharmacological properties. Therefore, the current study aimed to determine the dose-dependent role of GNK against AFB1-instigated hepatotoxicity. The investigation was carried out on 96 adult male albino rats, which were equally distributed into eight groups. The effect of 3 different doses of GNK (5, 10 and 20 mgkg-1) was evaluated against the toxicity elicited by 50 ugkg-1 of AFB1. After the administration of AFB1 and GNK by the oral gavage for 56 days, the biochemical and hepatic serum markers were determined in addition to histopathological observation. AFB1 exposure disrupted the biochemical profile by declining the activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione content), while elevating the concentration of reactive oxygen species and malondialdehyde level. Furthermore, AFB1 exposure notably elevated the levels of hepatic serum enzymes (alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase) along with the levels of inflammatory markers, nuclear factor kappa-B, tumor necrosis factor-α, Interleukin-6, Interleukin-1β and activity of cyclooxygenase-2. Besides, AFB1 induction caused histopathological impairments in hepatic tissues. Nonetheless, GNK co-administration remarkably ameliorated all the damages of the hepatic system induced by AFB1 administration to the rats. Therefore, it was demonstrated that the GNK could potentially cure AFB1-instigated hepatotoxicity attributing to its antioxidative and ant-inflammatory potential

Cyclooxygenase-2 Upregulated by Temozolomide in Glioblastoma Cells Is Shuttled In Extracellular Vesicles Modifying Recipient Cell Phenotype.

Temozolomide (TMZ) resistance is frequent in patients with glioblastoma (GBM), a tumor characterized by a marked inflammatory microenvironment. Recently, we reported that cyclooxygenase-2 (COX-2) is upregulated in TMZ-resistant GBM cells treated with high TMZ concentrations. Moreover, COX-2 activity inhibition significantly counteracted TMZ-resistance of GBM cells. Extracellular vesicles (EV) are considered crucial mediators in orchestrating GBM drug resistance by modulating the tumor microenvironment (TME) and affecting the surrounding recipient cell phenotype and behavior. This work aimed to verify whether TMZ, at low and clinically relevant doses (5-20 µM), could induce COX-2 overexpression in GBM cells (T98G and U87MG) and explore if secreted EV shuttled COX-2 to recipient cells. The effect of COX-2 inhibitors (COXIB), Celecoxib (CXB), or NS398, alone or TMZ-combined, was also investigated. Our results indicated that TMZ at clinically relevant doses upregulated COX-2 in GBM cells. COXIB treatment significantly counteracted TMZ-induced COX-2 expression, confirming the crucial role of the COX-2/PGE2 system in TMZ-resistance. The COXIB specificity was verified on U251MG, COX-2 null GBM cells. Western blotting of GBM-EV cells showed the COX-2 presence, with the same intracellular trend, increasing in EV derived from TMZ-treated cells and decreasing in those derived from COXIB+TMZ-treated cells. We then evaluated the effect of EV secreted by TMZ-treated cells on U937 and U251MG, used as recipient cells. In human macrophage cell line U937, the internalization of EV derived by TMZ-T98G cells led to a shift versus a pro-tumor M2-like phenotype. On the other hand, EV from TMZ-T98G induced a significant decrease in TMZ sensitivity in U251MG cells. Overall, our results, in confirming the crucial role played by COX-2 in TMZ-resistance, provide the first evidence of the presence and effective functional transfer of this enzyme through EV derived from GBM cells, with multiple potential consequences at the level of TME.

Celecoxib-mediated attenuation of non-alcoholic steatohepatitis is potentially relevant to redistributing the expression of adiponectin receptors in rats

Pharmacological inhibition of cyclooxygenase-2 (COX-2) activity ameliorated the severity of non-alcoholic steatohepatitis (NASH) rats. It is not completely understood that the role of COX-2 inhibitor celecoxib on adiponectin receptors (Adipo-R1/R2) expression in different tissues in NASH rats. Sprague-Dawley male NASH rats induced by a high-fat diet (HFD) were administrated with or without celecoxib for 8 weeks. Biochemical parameters of liver function, glucose, and lipid metabolism, and the levels of adiponectin, tumor necrosis factor-alpha (TNF-α), prostaglandin E2 (PGE2) in the serum or liver were collected according to the standard protocols. The mRNA and protein levels of Adipo-R1, Adipo-R2, and COX-2 in the liver, muscle, and visceral fat were performed by quantitative real-time polymerase chain reaction (q-PCR) and Western blot analysis, respectively. The results showed that celecoxib ameliorated the various clinical indicators and pathological characteristics in the NASH rats, including body weight, liver function, liver index, and redox activities in serum and hepatic samples. The serum concentrations of adiponectin and TNF-α and PGE2 were negatively correlated. As expected, these ameliorative effects of celecoxib were associated with the gene and protein levels up-regulation of Adipo-R1, Adipo-R2 in the liver and visceral fat tissues, and seeming to be compensatory down-regulation expression in muscle tissues (P <0.05). Additionally, COX-2 protein expression was negatively correlated with serum adiponectin levels, protein expression of adiponectin receptors from the liver and visceral fat, conversely, positively correlated with those from the muscle. Our current study demonstrate that celecoxib might effectively alleviate NASH rats in a unique manner closely relevant to redistributing the expression of adiponectin receptors in the liver, visceral fat, and muscle. However, the precise molecular mechanism needs further study.

Edwardsiella ictaluri T3SS Effector EseN Modulates Expression of Host Genes Involved in the Immune Response.

The type III secretion system (T3SS) effector EseN is encoded on the Edwardsiella ictaluri chromosome and is homologous to a family of T3SS effector proteins with phosphothreonine lyase activity. Previously we demonstrated that E. ictaluri invasion activates extracellular signal-regulated kinases 1 and 2 (ERK1/2) early in the infection, which are subsequently inactivated by EseN. Comparative transcriptomic analysis showed a total of 753 significant differentially expressed genes in head-kidney-derived macrophages (HKDM) infected with an EseN mutant (∆EseN) compared to HKDM infected with wild-type (WT) strains. This data strongly indicates classical activation of macrophages (the M1 phenotype) in response to E. ictaluri infection and a significant role for EseN in the manipulation of this process. Our data also indicates that E. ictaluri EseN is involved in the modulation of pathways involved in the immune response to infection and expression of several transcription factors, including NF-κβ (c-rel and relB), creb3L4, socs6 and foxo3a. Regulation of transcription factors leads to regulation of proinflammatory interleukins (IL-8, IL-12a, IL-15, IL-6) and cyclooxygenase-2 (COX-2) expression. Inhibition of COX-2 mRNA by WT E. ictaluri leads to decreased production of prostaglandin E2 (PGE2), which is the product of COX-2 activity. Collectively, our results indicate that E. ictaluri EseN is an important player in the modulation of host immune responses to E.ictaluri infection.

Influence of Inhibition of COX-2-Dependent Lipid Metabolism on Regulation of UVB-Induced Keratinocytes Apoptosis by Cannabinoids.

Inflammation and apoptosis are regulated by similar factors, including ultraviolet B (UVB) radiation and cannabinoids, which are metabolized by cyclooxygenase-2 (COX-2) into pro-apoptotic prostaglandin derivatives. Thus, the aim of this study was to evaluate the impact of cyclooxygenase-2 inhibition by celecoxib on the apoptosis of keratinocytes modulated by UVB, anandamide (AEA) and cannabidiol (CBD). For this purpose, keratinocytes were non-treated/treated with celecoxib and/or with UVB and CBD and AEA. Apoptosis was evaluated using microscopy, gene expressions using quantitate reverse-transcriptase polymerase chain reaction; prostaglandins using liquid chromatography tandem mass spectrometry and cyclooxygenase activity using spectrophotometry. UVB enhances the percentage of apoptotic keratinocytes, which can be caused by the increased prostaglandin generation by cyclooxygenase-2, or/and induced cannabinoid receptor 1/2 (CB1/2) expression. AEA used alone intensifies apoptosis by affecting caspase expression, and in UVB-irradiated keratinocytes, cyclooxygenase-2 activity is increased, while CBD acts as a cytoprotective when used with or without UVB. After COX-2 inhibition, UVB-induced changes are partially ameliorated, when anandamide becomes an anti-apoptotic agent. It can be caused by observed reduced generation of anandamide pro-apoptotic derivative prostaglandin-ethanolamide by COX. Therefore, products of cyclooxygenase-dependent lipid metabolism seem to play an important role in the modulation of UVB-induced apoptosis by cannabinoids, which is particularly significant in case of AEA as inhibition of cyclooxygenase reduces the generation of pro-apoptotic lipid mediators and thus prevents apoptosis.

Hepatoprotective role of vitexin against cadmium-induced liver damage in male rats: A biochemical, inflammatory, apoptotic and histopathological investigation.

Cadmium (Cd) is one of the potent occupational and environmental toxicants, which induces oxidative stress to the multiple organs of the body, including liver. The present investigation was planned to evaluate the protective role of vitexin against Cd-prompted hepatotoxicity in rats. 24 male rats were divided into 4 groups viz. control, Cd-induced group (5mg/kg), Cd +vitexin-treated group (2mg/kg+30mg/kg), and vitexin-treated group (30mg/kg). After 30 days of treatment, it was indicated that Cd escalated the level of liver function enzymes namely alanine transaminase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) as well as total bilirubin. Whereas the levels of albumin and total proteins were decreased in the rats. Additionally, it reduced the enzymatic activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GSR) and glutathione-S-transferase (GST), in addition to glutathione (GSH) content, whereas levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were escalated. Furthermore, level of nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) as well as the activity of cyclooxygenase-2 (COX-2) were increased. Besides, the level of Bax, caspase-9 and caspase-3 were elevated, while the Bcl-2 level was reduced following the Cd intoxication. Histopathological observation revealed significant hepatic tissue damage in Cd-administered rats. However, treatment of rats with vitexin significantly (p<0.05) improved the Cd-induced disruptions in biochemical parameters as well as histological damages. Therefore, it is concluded that vitexin could be used as a therapeutic agent to counter the Cd-generated hepatic toxicity in rats owing to its anti-oxidant, anti-apoptotic and anti-inflammatory potential.

Making a meal out of bugs

Making a meal out of bugs

Behavioral and inflammatory sex differences revealed by celecoxib nanotherapeutic treatment of peripheral neuroinflammation

Neuropathic pain affects millions of people worldwide, yet the molecular mechanisms of how it develops and persists are poorly understood. Given that males have historically been utilized as the primary sex in preclinical studies, less is known about the female neuroinflammatory response to injury, formation of pain, or response to pain-relieving therapies. Macrophages contribute to the development of neuroinflammatory pain via the activation of their cyclooxygenase-2 (COX-2) enzyme, which leads to the production of prostaglandin E2 (PGE2). PGE2 activates nociception and influences additional leukocyte infiltration. Attenuation of COX-2 activity decreases inflammatory pain, most commonly achieved by nonsteroidal anti-inflammatory drugs (NSAIDs), yet NSAIDs are considered ineffective for neuropathic pain due to off target toxicity. Using chronic constriction injury of the rat sciatic nerve, we show that males and females exhibit quantitatively the same degree of mechanical allodynia post injury. Furthermore, a low-dose nanotherapeutic containing the NSAID celecoxib is phagocytosed by circulating monocytes that then naturally accumulate at sites of injury as macrophages. Using this nanotherapeutic, we show that treated males exhibit complete reversal of hypersensitivity, while the same dose of nanotherapeutic in females provides an attenuated relief. The difference in behavioral response to the nanotherapy is reflected in the reduction of infiltrating macrophages at the site of injury. The observations contained in this study reinforce the notion that female neuroinflammation is different than males.

DPP4 Promotes Human Endothelial Cell Senescence and Dysfunction via the PAR2-COX-2-TP Axis and NLRP3 Inflammasome Activation.

Abnormal accumulation of senescent cells in the vessel wall leads to a compromised vascular function contributing to vascular aging. Soluble DPP4 (dipeptidyl peptidase 4; sDPP4) secretion from visceral adipose tissue is enhanced in obesity, now considered a progeric condition. sDPP4 triggers vascular deleterious effects, albeit its contribution to vascular aging is unknown. We aimed to explore sDPP4 involvement in vascular aging, unraveling the molecular pathway by which sDPP4 acts on the endothelium. Human endothelial cell senescence was assessed by senescence-associated β-galactosidase assay, visualization of DNA damage, and expression of prosenescent markers, whereas vascular function was evaluated by myography over human dissected microvessels. In visceral adipose tissue biopsies from a cohort of obese patients, we explored several age-related parameters in vitro and ex vivo. By a common mechanism, sDPP4 triggers endothelial cell senescence and endothelial dysfunction in isolated human resistance arteries. sDPP4 activates the metabotropic receptor PAR2 (protease-activated receptor 2), COX-2 (cyclooxygenase 2) activity, and the production of TXA2 (thromboxane A2) acting over TP (thromboxane receptor) receptors (PAR2-COX-2-TP axis), leading to NLRP3 (nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing 3) inflammasome activation. Obese patients exhibited impaired microarterial functionality in comparison to control nonobese counterparts. Importantly, endothelial dysfunction in obese patients positively correlated with greater expression of DPP4, prosenescent, and proinflammatory markers in visceral adipose tissue nearby the resistance arteries. Moreover, when DPP4 activity or sDPP4-induced prosenescent mechanism was blocked, endothelial dysfunction was restored back to levels of healthy subjects. These results reveal sDPP4 as a relevant mediator in early vascular aging and highlight its capacity activating main proinflammatory mediators in the endothelium that might be pharmacologically tackled.

Protective effects of aucubin against nonylphenol-induced liver toxicity by improving biochemical, inflammatory and histopathological indices

Nonylphenol (NP) is a ubiquitous environmental toxicant that is known to cause hepatotoxicity via generation of reactive oxygen species (ROS). Aucubin (AUC), an iridoid glucoside, is a potential phytochemical possessing antioxidant and anti-inflammatory features. The current study was intended to ascertain the hepatoprotective role of AUC against NP-generated liver toxicity in rats. 48 male rats were distributed into four groups. viz. Control, NP-intoxicated group (50 mg/kg), NP + AUC-treated group (50 mg/kg + 40 mg/kg) and AUC-treated group (40 mg/kg). All the doses were administered orally. Our findings indicated that the NP intoxication upregulated the alanine aminotransferase (ALT), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) level. Moreover, it brought down the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GSR) and glutathione-S-transferase (GST), as well as the glutathione (GSH) content. Conversely, the levels of malondialdehyde (MDA) and ROS were escalated. Besides, the levels of nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 as well as the cyclooxygenase-2 (COX-2) activity were elevated due to the NP administration. In addition to it, histopathological assessment displayed the prominent morphological alterations in hepatic tissues of rats. Nevertheless, treating the rats with AUC significantly (p < 0.05) abated all the NP-instigated liver damages in rats. Therefore, it was evinced that AUC could be used as a hepatoprotective agent against the NP-prompted liver toxicity owing to its antioxidant and anti-inflammatory properties.

Carvacrol Arrests the Proliferation of Hypopharyngeal Carcinoma Cells by Suppressing Ornithine Decarboxylase and Hyaluronidase Activities.

Carvacrol, a monoterpene known for its pharmacological activities, is present in the essential oil of Origanum majorana, Origanum vulgare, Thymus vulgaris, and Lippia graveolens. It is used in food as a flavoring and preservative agent in cosmetics and medicines because of its useful bioactivities in clinical practice. However, carvacrol was not much explored for its anticancer potential. Targeting enzymes involved in carcinogenesis, such as ornithine decarboxylase (ODC), cyclooxygenase-2 (COX-2), lipoxygenase-5 (LOX-5), and hyaluronidase (HYAL) by monoterpenes are amongst the efficient approaches for cancer prevention and treatment. In this study, the efficacy of carvacrol was investigated against deregulated cancer biomarkers/targets in organ-specific human cancer cell lines (FaDu, K562, and A549) utilizing in vitro, in silico, and in vivo approaches. The efficacy of carvacrol was evaluated on human cancer cell lines using neutral red uptake (NRU), sulpho rhodamine B (SRB), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. The mechanistic study was carried out in cell-based test systems. Further, the potency of carvacrol was confirmed by the quantitative real-time PCR analysis and molecular docking studies. The in vivo anti-tumor potential of carvacrol was performed on mice S-180 model, and the toxicity examination was accomplished through in silico approach. Carvacrol significantly impeded the growth of FaDu, K562, and A549 cell lines with IC50 values ranging from 9.61 ± 0.05 to 81.32 ± 11.83 μM. Further, the efficacy of carvacrol was explored against different cancer targets in FaDu, K562, and A549 cell lines. Carvacrol inhibits the ODC, COX-2, LOX-5, and HYAL activities in FaDu cell line and ODC, COX-2, and HYAL activities in K562 cell line. The results were validated by expression analysis revealing the downregulation of the targeted gene with a significant change in the transcript level of ODC and HYAL in FaDu cell line with a fold change of 1.56 and 1.61, respectively. A non-significant effect of carvacrol was observed on the downstream signaling pathway of PI3K and HIF-1α/vascular endothelial growth factor (VEGF) in FaDu cells. The cell cycle, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and Annexin V-fluorescein isothiocyanate (FITC) experiments demonstrate that carvacrol induces apoptosis of FaDu cells. Further, the potency of carvacrol was also evaluated in vivo on mice S-180 tumor model, wherein it inhibits tumor growth (72%) at 75 mg/kg body weight (bw). ADMET studies predicted carvacrol as a safe molecule. Overall, carvacrol delayed the growth of FaDu, K562, and A549 cell lines by targeting enzymes involved in the carcinogenesis process. The existence of one hydroxyl group at the para position of carvacrol could be responsible for the anti-proliferative activity. Thus, carvacrol could be used as a pharmacophore to develop a safe and effective multi-targeted anti-cancer medicament.