Undescribed C-Glycosylflavones from Corn Silk and Potential Anti-inflammatory Activity Evaluation of Isolates

Phytochemical investigation of corn silk resulted in isolation and characterization of four flavone C-glycosides, chrysoeriol 6-C-β-oliopyranosyl-7-O-β-D-glucopyranoside (1), 3′-methoxycassiaoccidentalin A (2), chrysoeriol 6-C-β-boivinopyranosyl-7-O-β-D-glucopyranoside (3), and ax-4"-OH-3′-methoxymaysin (4), a triterpenoid, friedelin (5), two sterols, (22E)-5,8-epidioxyergosta-6,22-dien-3β-ol (6) and 6β-hydroxystigmasta-4,22-diene-3-one (7), and a mixture of β-sitosterol and stigmasterol. Compounds 1 and 2 were previously undescribed. Structure elucidation of the isolated compounds was attained using spectral data including 1D and 2D NMR and HRESIMS. Compounds 1, 2, 5, and 6 inhibited iNOS activity in LPS-induced macrophages and decreased nitrite levels by 68.64+4.46, 65.67 + 6.47, 88.50 + 0.50, and 94.00 + 4.00 %, respectively, at 50 µM. Compound 5 also showed inhibition of NF-κB (51.00+1.50 %). Compounds 1 and 2 induced NAG-1 activity in chondrocytes by 1.80 + 0.05 and 2.00 + 0.13 fold, respectively. The extract of corn silk, however, did not exhibit inhibition of iNOS or NF-κB but induced NAG-1 by 1.80+ 0.51 fold.

Oleuropein-Rich Leaf Extract as a Broad Inhibitor of Tumour and Macrophage iNOS in an Apc Mutant Rat Model

Oleuropein, the major compound found in olive leaves, has been reported to exert numerous pharmacological properties, including anti-inflammatory, anti-diabetic and anti-cancer effects. The purpose of this study was to evaluate, for the first time, the effect of oleuropein-rich leaf extracts (ORLE) in already-developed colon tumours arising in Apc (adenomatous polyposis coli) mutated PIRC rats (F344/NTac-Apcam1137). Here, we were able to investigate in parallel the anti-cancer effect of ORLE, both in vivo and in vitro, and its anti-inflammatory effect on macrophages, representing a critical and abundant population in most solid tumour microenvironment. We found that in vivo ORLE treatment promoted apoptosis and attenuated iNOS activity both in colon tumours as in peritoneal macrophages of PIRC rats. We this confirmed in vitro using primary RAW264.7 cells: ORLE reduced iNOS activity in parallel with COX-2 and pro-inflammatory cytokines, such as IL-1β, IL-6 and TGF-β. These findings suggest that ORLE possess a strong anti-inflammatory activity, which could be crucial for dampening the pro-tumourigenic activity elicited by a chronic inflammatory state generated by either tumour cells or tumour-associated macrophages.

Eicosapentaenoic acid inhibits lipopolysaccharide (LPS)-induced nitrite production and cytokine release from J774 macrophages

Background Eicosapentaenoic acid (EPA), an omega-3 (ω-3) fatty acid, reduced cardiovascular (CV) events in high-risk patients (REDUCE-IT) but the mechanism is not fully understood. Activated macrophages, characterized by cytokine release and increased inducible nitric oxide synthase (iNOS) activity, contribute to atherosclerosis. As both a substrate for and potential inhibitor of cyclooxygenase (COX), EPA may reduce iNOS activity. Purpose The purpose of this study was to evaluate the dose-dependent effects of EPA on nitrite and cytokine release from lipopolysaccharide (LPS)-activated macrophages. Methods Murine J774 macrophages were pretreated with vehicle or EPA at 10, 20 and 40 μM for 2 h, then challenged with LPS at 1.0 μg/ml. After 24 hr, iNOS activity was measured by nitrite production using the Griess assay. EPA was compared to the COX inhibitor diclofenac at 1.0 μg/ml. Levels of interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) in cell supernatant were measured by immunochemistry using colchicine as a positive control. Results Activated macrophages caused a >4-fold increase in nitrite production (p<0.001) that was reduced by EPA in a dose-dependent manner. EPA decreased nitrite levels by 40, 62 and 77% at 10, 20 and 40 μM, respectively (p<0.01). Diclofenac separately reduced nitrite levels by 40% (p<0.01). EPA also reduced expression of IL-1β and TNF-α by 40% and 31%, respectively (p<0.01), in a manner similar to equimolar colchicine (10 μM). The reductions in IL-1β and TNF-α with EPA were dose-dependent. Conclusions EPA reduced macrophage activation as evidenced by decreased nitrite production and cytokine release similar to other anti-inflammatory agents. These findings indicate a novel effect of EPA on mechanisms of inflammation associated with vascular disease. FUNDunding Acknowledgement Type of funding sources: Private company. Main funding source(s): Amarin Pharma Inc., Elucida Research

COX2: A Prognostic Inflammogenic Marker Drives Cervical Carcinogenesis in Vivo through the NFκB/IAP/p53Axis

Cycloxygenase 2, a prostaglandin synthesizing enzyme, is a key player in inflammation-induced vasculogenesis that enables tumor growth. This study explores the central role of COX2 and its relative prosurvival proteins in evoking inflammatory events during the development of an in vivo cervical cancer model upon chronic treatment with 3-methylcholanthrene (3MC; a chemical carcinogen) in virgin-female Swiss albino mice. Chronic painting of the mouse cervix with 3MC solution triggered the persistent expression and activity of COX2, eventuating the overexpression of major prosurvival molecules (NFκB, XIAP, survivin, GM-CSF1) and proliferative antigens (Ki67, PCNA). COX2-arbitrated prosurvival signaling subsequently deranged the expression profiles of tumor suppressor proteins (p53/acetyl-p53, p21, Rb) within the cervix. COX2 mediated molecular alterations successively surged leukocyte influx within the cervix, catering to localized inflammation that gradually distorted its tissue architecture. Cervical carcinogenesis was further braced by higher levels of systemic ROS and RNS, escalated iNOS activity and compromised antioxidant enzyme capacities, which were accompanied by splenomegaly. Additionally, circulation of blood leucocytes with damaged DNA throughout the mouse body envisaged the impact of cervix-limited inflammation on mouse physiology. Conclusively, the present study deciphered the role of COX2 in affecting NFκB/IAP/p53 functions in sequestering the contributors of localized and systemic inflammogenesis to propel 3MC-mediated cervical carcinogenesis in vivo.

The role of the macrophage polarization type in the pathogenesis of endometrioid disease

Endometriosis today occupies one of the leading places in the structure of general gynecological pathology. Theories of the onset and progression of this disease are controversial. One of the most widespread theories is the assumption that endometriosis is a disease of macrophages. The question of which the macrophage phenotype, M1 or M2, is the leading one, however, remains controversial. The aim. To determine the type of macrophage polarization (M1/M2) and the quantitative activity of their marker enzymes (iNos/Arg1) in the endometrium and peritoneal fluid in endometrioid disease. Materials and methods. The total number of reproductive age (30.95±6.49) women enrolled in the study was 80. The main group consisted of 50 women with endometrioid disease. The control group included 30 women without signs of endometrioid disease. Women from the main group (n=24) and the control group (n=27) underwent endometrial sampling using an intrauterine Pipelle catheter in the first phase of the menstrual cycle before a surgery. During laparoscopic or laparotomy approaches, peritoneal fluid was taken (in the main group n=24, in the control group n=28). The type of macrophage polarization (M1 or M2) was determined based on the ratio of marker enzymes (Arg1, iNos) activity in each patient using a spectrophotometric method in the endometrium and peritoneal fluid. The polarization to the M1 phenotype was determined at iNos>Arg1, and at Arg1>iNos– the polarization to the M2 phenotype. Results. As a result of the study, it was revealed that in women with endometrioid disease, pelvic adhesions were much more common, 84.0% versus 46.7% in women without it, and especially 3 and 4 degree of severity (P<0.05). When assessing the type of macrophages in the peritoneal fluid, a significantly greater number of the main group women had the M2 phenotype of macrophage polarization compared to the control group (58.3% versus 28.6%, P=0.03). It was the macrophage polarization to the M2 phenotype that influenced the severity of endometrioid disease, especially the 4 degree of severity. The mean values of the iNOS activity in the main group women, both in the peritoneal fluid and in the endometrium, significantly differed from those in the control group patients (by 1.73 and 1.77 times, respectively). Conclusions. Thus, we can conclude that endometriosis is a disease, the development and progression of which is induced by the M2 phenotype of macrophages. Considering the increase in the mean levels of iNOS activity both in the peritoneal fluid and in the endometrium, it can be concluded that iNOS influences the pathogenesis of endometrioid disease.

Oleuropein-Rich Leaf Extract as a Broad Inhibitor of Tumour and Macrophage iNOS in Apc Mutant Rat Model

Oleuropein, the major compound of olive leaves, has been reported to exert numerous pharmacological properties, including anti-inflammatory, antidiabetic and anticancer. The purpose of this study is to evaluate, for the first time, the effect of oleuropein-rich leaf extracts (ORLE) in already-developed colon tumours colon tumours arising in an Apc (adenomatous polyposis coli) mutated PIRC rats (F344/NTac-Apcam1137). Here, we were able to investigate in parallel the anti-cancer effect of ORLE, both in vivo and in vitro, and its anti-inflammatory effect on macrophages, which represents a critical and abundant population in most solid tumours microenvironment. We found that in vivo ORLE treatment promoted apoptosis and attenuated iNOS activity both in colon tumours as in peritoneal macrophages of PIRC rats. We confirmed in vitro using primary RAW264.7 cells: ORLE reduced iNOS activity in parallel with COX-2 and pro-inflammatory cytokines, such as IL-1, IL-6 and TGF-. These findings suggest that ORLE possess a strong anti-inflammatory activity, which could be crucial for dampening the pro-tumourigenic activity elicited by a chronic inflammatory state generated by either tumour cells or tumour-associated macrophages.

The Effect of Cinnamaldehyde on iNOS Activity and NO-Induced Islet Insulin Secretion in High-Fat-Diet Rats

Introduction. Obesity and insulin resistance are associated with alterations in nitric oxide level and insulin secretion. Previous studies demonstrated that cinnamaldehyde (CNMA) improved islet insulin secretion and restored nitric oxide (NO) level, but its underlying mechanisms have not been investigated. This study aimed to investigate the effect of CNMA on inducible nitric oxide synthase (iNOS) activity and NO-induced islet insulin secretion in high-fat-diet (HFD) treated rats. Materials and Methods. Forty male Wistar rats (12 weeks old) were randomly divided into four equal groups, namely, control, CNMA, HFD, and HFD + CNMA. Control and CNMA groups were treated with standard laboratory animals’ diet, while HFD and HDF + CNMA groups were fed with an HFD diet enriched with 25% W / W tail fat for 16 weeks. CNMA was administrated orally (20 mg/kg body weight, daily) during the study period. Islet insulin secretion and the inducible NOS activity in the presence or absence of L-NAME (NO synthase inhibitor, 5 mmol/L) were evaluated. Results. L-NAME-suppressed insulin secretion in control, HFD, and HFD + CNMA groups; however, in the CNMA group, it could not exhibit such effect ( P < 0.01 ). Islets of HFD-treated animals showed significantly higher iNOS activity than controls. CNMA treatment significantly suppressed iNOS activities in CNMA and HFD + CNMA groups compared with control and HFD, respectively. Conclusion. These results suggest that the beneficial effect of CNMA on insulin secretion might be due to its inhibitory effect on iNOS activity.

Posible differential diagnosis of various chronic nonbacterial prostatites

Background: The purpose of the study was to diagnose possible chronic nonbacterial prostatitis (CNP) and chronic pelvic pain syndrome (CPPS) among patients, as well as differentiate between the inflammatory (category IIIA) or non-inflammatory (category IIIB) types in selecting and optimizing differential drug treatment of this category of patients. Material and methods: The study was conducted on 43 patients diagnosed with CNP/CPPS. The control group included 10 healthy men. Both the production of nitric oxides (NO) by phagocytes, as well as prostate secretion and ejaculate were determined according to the procedure described by Metelyskaya B.A., which was modified by Gudumac V, et al. Results: There was a 39.0% (p <0. 05) decrease in NO production by induced NO-synthase (iNOS), determined in the blood of 11 patients (from the main group – 2) with CNP/CPPS and a 115% (p <0.05) increase was determined in 32 patients (from the main group 1) if compared to the same indices in the control group. The prostatic secretion and ejaculate showed a higher macrophage iNOS activity by 80% (p <0.05) and 75% (p <0.05) if compared to the same parameters from the control group. The iNOS activity in prostatic fluid and split-ejaculate fractions from the main group – 2 did not differ from that of the control group. Conclusions: The assessment of NO production, prostate secretion and ejaculate allows to somewhat establish the main diagnosis of CNP and category III types (A – inflammatory and B – non-inflammatory prostatitis), which will significantly contribute to the optimization and selection of an appropriate differential treatment based on the drug action mechanisms