SLN and chitosan nano-delivery systems for antibacterial effect of black seed (Nigella sativa) oil against S. aureus.

Staphylococcus aureus with current universal importance represents a main carrier of emerging antimicrobial resistance determinatives of global health concerns that have developed drug resistance mechanisms to the various available antibiotics. On the other hand, due to the antimicrobial potential of Nigella Sativa oil (NSO), it was hypothesized that incorporation of nano-carriers (NS-SLN and NS-chitosan (CH) nanoparticles) can enhance its antibacterial effects. This study evaluated the physico-chemical and antibacterial characteristics of NS-SLN and NS-CH. TEM images revealed a round shape with clear edges for both nanoparticles, and the average sizes were reported to be 196.4 and 446.6 nm for NS-SLN and NS-CH, respectively. The zeta potential and encapsulation efficiency were -28.9 and 59.4 mV and 73.22% and 88% for NS-SLN and NS-CH, respectively. The Minimum Inhibitory Concentrations for NSO, NS-SLN, and NS-CH against S. aureus were 480, 200, and 80 µg/mL, respectively. The results confirm significantly stronger antibacterial influences of NSO when loaded into chitosan nanoparticles as a potential candidate for nano-delivery of antimicrobial agents.

The effects of high doses of selenium supplementation on mRNA and protein levels of cMLCK levels and total antioxidant capacity in rat heart tissue.

BACKGROUND: High doses of selenium are associated with heart disease prevalence in high-risk areas. Cardiac myosin light chain kinase (cMLCK) is an essential enzyme for normal function of heart tissue. Therefore, we studied the effect of high doses of selenium on the expression of cMLCK gene and its protein in normal heart tissue in rats. MATERIALS AND METHODS: Twenty male rats were randomly divided into four groups: control, Se 0.3mg/kg, Se 1.5mg/kg, and Se 3mg/kg. Sodium-selenite was administered orally into drinking water for 20 weeks. Se levels of heart tissue were measured by atomic absorption. Serum creatine phosphokinase (CPK) and total serum antioxidant capacity were measured. Moreover, the concentration of MLCK protein and the gene expression level of cMLCK in normal heart tissue were analyzed. RESULTS: Excess Se in dietary can significantly increase CPK. Se concentration of heart tissue in the Se 3mg/kg group was significantly higher than the control. cMLCK mRNA levels were decreased by 0.3mg/kg and 3mg/kg sodium selenite intake. There was no significant difference between the three groups for total antioxidant capacity and MLCK protein. CONCLUSION: High concentrations of selenium can probably effect on normal function of the heart tissue by changing the expression levels of cMLCK.

Evaluation of the Effect of Pistacia atlantica Oleoresin on Blood Sugar, Pressure and Lipids in Patients With Type 2 Diabetes: A Single-Blind, Placebo-Controlled Trial.

BACKGROUND: Diabetes mellitus (DM) is a chronic metabolic disorder characterised by high blood sugar (BS) levels due to impaired insulin production or insulin resistance. It is a global health concern with significant implications for morbidity and mortality. Persian medicine has long utilised natural remedies, such as Pistacia atlantica Desf., for various diseases. In this randomised clinical trial, the effects of P. atlantica oleoresin in the improvement of lipid profiles, glucose indices and blood pressure (BP) were assessed in patients with Type 2 DM. MATERIALS AND METHODS: In this randomised, single-blind, placebo-controlled study, 42 patients with Type 2 DM were randomly allocated to receive either P. atlantica oleoresin or placebo capsule for 3 months. Patients were evaluated prior to and 12 weeks after the beginning of the intervention, in terms of changes in lipid profiles, glucose indices and BP. RESULTS: After 3 months, the mean BP in patients with DM receiving P. atlantica oleoresin was significantly reduced compared with the baseline (p = 0.001). Also, these changes were significantly higher than those of the control group. The mean of total cholesterol (p = 0.89), low-density lipoprotein (LDL) (p = 0.43) and triglyceride (TG) (p = 0.98) in the intervention group after 3 months was lower than that in the control group, but this difference was not statistically significant. CONCLUSION: After 3 months, there was no significant difference between the P. atlantica and control groups in terms of blood sugar and lipid profiles. The mean BP in patients with DM receiving P. atlantica oleoresin was significantly reduced compared with that in the beginning of the study. Also, these changes were significant compared with the control group.

Anti-Toxoplasma gondii activity of rose hip oil-solid lipid nanoparticles.

Toxoplasma gondii is a highly prevalent pathogen, reported from almost all geographical regions of the world. Current anti-T. gondii drugs are not effective enough in immunocompromised patients, encephalitis, chorioretinitis, and congenital toxoplasmosis. Therefore, the prescription of these drugs has been limited. Rose hip oil (RhO) is a natural plant compound, which shows antibacterial, anticancer, and anti-inflammatory activities. In the current study, the anti-T. gondii and cell toxicity effects of solid lipid nanoparticles (SLNs) loaded by RhO (RhO-SLNs) were evaluated. Emulsification sonicated-homogenization method was used to prepare SLNs. RhO-SLNs were characterized, and their anti-T. gondii and cell toxicity effects were evaluated using in vitro analyses. The particle size and the zeta potential of the nanoparticles were 152.09 nm and -15.3 mV nm, respectively. The entrapment efficiency percentage was 79.1%. In the present study, the inhibitory concentration (IC)50 against T. gondii was >1 µg/mL (p-value <.0001). The cell toxicity assay showed cytotoxicity concentration (CC)50 >10 mg/mL (p-value = .017). In addition, at least 75% of T. gondii-infected Vero cells remained alive at concentrations >10 mg/mL. The concentration of 1 mg/mL showed highest anti-Toxoplasma activity and lowest cell toxicity against the Vero cell. Our findings suggest that carrying natural plant compounds with SLNs could be considered an effective option for treatment strategies against T. gondii infections.

Development and characterization of a topical gel, containing lavender (Lavandula angustifolia) oil loaded solid lipid nanoparticles.

Gels loaded with nanocarriers offer interesting ways to create novel therapeutic approaches by fusing the benefits of gel and nanotechnology. Clinical studies indicate that lavender oil (Lav-O) has a positive impact on accelerating wound healing properly based on its antimicrobial and anti-inflammatory effects. Initially Lav-O loaded Solid Lipid Nanoparticles (Lav-SLN) were prepared incorporating cholesterol and lecithin natural lipids and prepared SLNs were characterized. Next, a 3% SLN containing topical gel (Lav-SLN-G) was formulated using Carbopol 940. Both Lav-SLN and Lav-SLN-G were assessed in terms antibacterial effects against S. aureus. Lav-SLNs revealed a particle size of 19.24 nm, zeta potential of -21.6 mv and EE% of 75.46%. Formulated topical gel presented an acceptable pH and texture properties. Minimum Inhibitory/Bactericidal Concentration (MIC/MBC) against S. aureus for LAv-O, Lav-SLN and Lav-SLN-G were 0.12 and 0.24 mgml- 1, 0.05 and 0.19 mgml- 1 and 0.045, 0.09 mgml- 1, respectively. Therefore, SLN can be considered as an antimicrobial potentiating nano-carrier for delivery of Lav-O as an antimicrobial and anti-inflammatory agent in topical gel.

The route of autophagy regulation by osteopontin: a review on the linking mechanisms.

Autophagy is a dynamic intracellular process of protein degradation, which is mostly triggered by nutrient deprivation. This process initiates with the formation of autophagosomes, which they capture cytosolic material that is then degraded upon fusion with the lysosome. Several factors have been found to be associated with autophagy modulation, of which extracellular matrix (ECM) components has attracted the attention of recent studies. Osteopontin (OPN) is an important extracellular matrix component that has been detected in a wide range of tumor cells, and is involved in cancer cell invasion and metastasis. Recently, a number of studies have focused on the relationship of OPN with autophagy, by delineating the intracellular signaling pathways that connect OPN to the autophagy process. We will summarize signaling pathways and cell surface receptors, through which OPN regulates the process of autophagy.

Comparison of chitosan and SLN nano-delivery systems for antibacterial effect of tea tree (Melaleuca alternifolia) oil against P. aeruginosa and S. aureus.

Treatment of wounds is challenging due to bacterial infections, including Staphylococcus aureus and Pseudomonas aeruginosa. Using the merits of alternative antimicrobials like tea tree oil (TTO) and nanotechnology, they can be helpful in combatting bacterial infections. Solid lipid nanoparticle (SLN) and chitosan (CS) nanoparticles show great potential as carriers for enhancing the stability and therapeutic benefits of oils. The aim of this study is to compare the influence of nanocarriers in enhancing the antibacterial effects of TTO. The study evaluates the physicochemical and antibacterial properties of TTO-SLN and TTO-CS against P. aeruginosa and S. aureus. The TTO-SLN nanoparticles showed a clear round shape with the average diameter size of 477 nm, while the TTO-CS nanoparticles illustrated very homogeneous morphology with 144 nm size. The encapsulation efficiency for TTO-CS and TTO-SLN was ∼88.3% and 73.5%, respectively. Minimum inhibitory concentration against S. aureus and P. aeruginosa for TTO-CS, TTO-SLN, and pure TTO were 35 and 45 µg ml-1, 130 and 170 µg ml-1, and 380 and 410 µg ml-1, respectively. Since TTO-CS revealed an impressively higher antimicrobial effects in comparison with TTO-SLN and TTO alone, it can be considered as a nanocarrier that produces the same antimicrobial effects with lower required amounts of the active substance.

The role of NFATc1 in the progression and metastasis of prostate cancer: A review on the molecular mechanisms and signaling pathways.

A common type of cancer among men is the prostate cancer that kills many people every year. The multistage of this disease and the involvement of the vital organs of the body have reduced the life span and quality of life of the people involved and turned the treatment process into a complex one. NFATc1 biomarker contributes significantly in the diagnosis and treatment of this disease by increasing its expression in prostate cancer and helping the proliferation, differentiation, and invasion of cancer cells through different signaling pathways. NFATc1 is also able to target the metabolism of cancer cells by inserting specific oncogene molecules such as c-myc that it causes cell growth and proliferation. Bone is a common tissue where prostate cancer cells metastasize. In this regard, the activity of NFATc1, through the regulation of different signaling cascades, including the RANKL/RANK signaling pathway, in turn, increases the activity of osteoclasts, and as a result, bone tissue is gradually ruined. Using Silibinin as a medicinal plant extract can inhibit the activity of osteoclasts related to prostate cancer by targeting NFATc. Undoubtedly, NFATc1 is one of the effective oncogenes related to prostate cancer, which has the potential to put this cancer on the path of progression and metastasis. In this review, we will highlight the role of NFATc1 in the progression and metastasis of prostate cancer. Furthermore, we will summarize signaling pathways and molecular mechanism, through which NFATc1 regulates the process of prostate cancer.

Formulation of Cinnamon (Cinnamomum verum) oil loaded solid lipid nanoparticles and evaluation of its antibacterial activity against Multi-drug Resistant Escherichia coli.

Today, the increment in microbial resistance has guided the researches focus into new antimicrobial compounds or transmission systems. Escherichia coli (E. coli) is an opportunistic pathogen, producing a biofilm responsible for a wide range of nosocomial infections which are often difficult to eradicate with available antibiotics. On the other hand, Cinnamomum verum (cinnamon oil) (CO) is widely used as a natural antibacterial agent and Solid lipid nanoparticles (SLNs) are promising carriers for antibacterial compounds due to their lipophilic nature and ease of transmission through the bacterial cell wall. In this study, nanoparticles containing cinnamon oil (CO-SLN) were prepared by dual emulsion method and evaluated in terms of particle size, shape, entrapment efficiency (EE), transmission electron microscopy (TEM), oil release kinetics, and cell compatibility. The antibacterial activity of CO-SLN and CO against 10 drug-resistant E. coli strains was investigated. The anti-biofilm activity of CO-SLN on the selected pathogen was also investigated. Nanoparticles with an average size of 337.6 nm, and zeta potential of -26.6 mV were fabricated and their round shape was confirmed by TEM images. The antibacterial effects of CO-SLN and CO were reported with MIC Value of 60-75 µg/mL and 155-165 µg/mL and MBC value of 220-235 µg/ml and 540-560 µg/ml, respectively. On the other hand, CO-SLN with 1/2 MIC concentration had the greatest inhibition of biofilm formation in 24 h of incubation (55.25%). The data presented indicate that the MIC of CO-SLN has significantly reduced and it seems that SLN has facilitated and promoted CO transmission through the cell membrane.

Formulation of Neem oil-loaded solid lipid nanoparticles and evaluation of its anti-Toxoplasma activity.

BACKGROUND: Toxoplasmosis is caused by an intracellular zoonotic protozoan, Toxoplasma gondii, which could be lethal in immunocompromised patients. This study aimed to synthesize Neem oil-loaded solid lipid nanoparticles (NeO-SLNs) and to evaluate the anti-Toxoplasma activity of this component. METHODS: The NeO-SLNs were constructed using double emulsification method, and their shape and size distribution were evaluated using transmission electron microscope (TEM) and dynamic light scattering (DLS), respectively. An MTT assay was employed to evaluate the cell toxicity of the component. The anti-Toxoplasma activity of NeO-SLNs was investigated using vital (trypan-blue) staining. Anti-intracellular Toxoplasma activity of NeO-SLNs was evaluated in T. gondii-infected Vero cells. RESULTS: The TEM analysis represented round shape NeO-SLNs with clear and stable margins. DLS analysis showed a mean particle size 337.6 nm for SLNs, and most of nanoparticles were in range 30 to 120 nm. The cell toxicity of NeO-SLNs was directly correlated with the concentration of the component (P-value = 0.0013). The concentration of NeO-SLNs, which was toxic for at least 50% of alive T. gondii (cytotoxic concentration (CC50)), was > 10 mg/mL. The ability of NeO-SLNs to kill Toxoplasma was concentration-dependent (P-value < 0.0001), and all concentrations killed at least 70% of alive tachyzoites. Furthermore, the viability of T. gondii- infected Vero cells was inversely correlated with NeO-SLNs concentrations (P-value = 0.0317), and in the concentration 100 µg/mL at least 75% of T. gondii- infected Vero cells remained alive. CONCLUSIONS: Overall, our findings demonstrated that the NeO-SLNs was able to kill T. gondii tachyzoites in concentration 100 µg/mL with a cell toxicity lower than 20%. Such results suggest that employing SLNs as carrier for NeO can effectively kill T. gondii tachyzoites with acceptable cell toxicity. Our findings also showed that SLNs capsulation of the NeO can lead to prolonged release of the extract, suggesting that NeO-SLNs could be also employed to clear cyst stages, which should be further investigated in animal models.

SPION based magnetic PLGA nanofibers for neural differentiation of mesenchymal stem cells.

Recently, magnetic platforms have been widely investigated in diagnostic, therapeutic and research applications due to certain properties, such as cell and tissue tracking and imaging, thermal therapy and being dirigible. In this study, the incorporation of magnetic nanoparticles (MNPs) in nanofibers has been proposed to combine the advantages of both nanofibers and MNPs to induce neural differentiation of mesenchymal stem cells. Magnetic poly (lactic-co-glycolic acid) nanofibers (containing 0%, 5% and 10% SPION) were fabricated and utilized as the matrix for the differentiation of mesenchymal stem cells (MSCs). Morphological, magnetic and mechanical properties were analyzed using FESEM, VSM and tensile test, respectively. The expression of neural markers (TUJ-1, NSE, MAP-2) was assessed quantitative and qualitatively utilizing RT-PCR and immunocytochemistry. Results confirmed the incorporation of MNPs in nanofibrous scaffold, presenting a saturation magnetization of 9.73 emu g-1. Also, with increase in magnetic particle concentration (0%-10%), tensile strength increased from 4.08 to 5.85 MPa, whereas the percentage of elongation decreased. TUJ-1 expression was 3.8 and 1.8 fold for 10% and 5% magnetic scaffold (versus non-magnetic scaffold) respectively, and the expression of NSE was 6.3 and 1.2-fold for 10% and 5%, respectively. Consequently, it seems that incorporation of magnetic biomaterial can promote the neural differentiation of MSCs, during which the augmentation of super paramagnetic iron oxide concentration from 0% to 10% accelerates the neural differentiation process.

Plasma Changes of Branched-Chain Amino Acid in Patients with Esophageal Cancer.

BACKGROUND Studies have indicated that branched amino acids play a crucial role in gene expression, protein metabolism, apoptosis, and restoration of hepatocytes and insulin resistance. This study aimed to compare the plasma levels of branched-chain amino acids in patients with esophageal cancer and normal individuals. METHODS Plasma levels of leucine and isoleucine of 37 patients with esophageal cancer and 37 healthy adults were investigated by high-pressure liquid chromatography. Data analysis was performed using SPSS (version 16) software, and t test was used to compare the plasma levels of branched-chain amino acids in the two groups. RESULTS In the patients group, the mean age ± SD was 63 ± 13.64 years, and 21 (56.8%) individuals were male. In the control group, the mean age ± SD was 64.24 ± 13.08 years, and 21 (54.1%) individuals were male. Plasma levels of leucine (37.68 ± 105) and isoleucine (22.43 ± 59.1) in patients with esophageal cancer were significantly reduced (p value of isoleucine:0.007, and leucine: 0.0001). CONCLUSION In the present study, the plasma levels of branched-chain amino acids in patients with esophageal cancer had changed. Evidence suggests that branched-chain amino acids are essential nutrients for cancer growth and are used by tumors in various biosynthetic pathways as energy sources. Thus, studies in this field can be useful in providing appropriate therapeutic approaches.

MiR-613 Promotes Cell Death in Breast Cancer Cells by Downregulation of Nicotinamide Phosphoribosyltransferase and Reduction of NAD.

NAD is mainly biosynthesized by the enzymatic action of nicotinamide phosphoribosyltransferase (NAMPT) through the salvage pathway. NAD is indispensable for the proper function and metabolism of all living cells, including cancer cells. Our previous researches revealed that inhibition of NAMPT by miRNA (miR) could suppress NAD levels and thereby hinder the growth and promotion of breast cancer (BC). Therefore, the current study was undertaken to investigate the inhibitory effects of miR-613 on NAMPT and BC cells' survival. Bioinformatics analysis and luciferase reporter assay confirmed that NAMPT 3'-untranslated region is a direct target for miR-613. The expression of miR-613 was noticed to be significantly decreased in both clinical tissue samples and BC cells by real-time PCR. Following transfection with miR-613 mimic, the expression of miR-613 was elevated in the BC cells leading to inhibition of NAMPT expression at both mRNA and protein level as measured by real-time PCR and western blotting, respectively. Inhibition of NAMPT led to a remarkable reduction in the concentration of NAD in the BC cells. The transfection also declined cell viability roughly 40% in MD Anderson-Metastatic Breast-231 (MDA-MB-231) cells. Consistently, the apoptosis rate was remarkably increased, around 65% in these cells as assayed by labeling the cells with Annexin V-fluorescein isothiocyanate (FITC) and Propidium Iodide. Targeting the NAMPT-mediated NAD salvage pathway by miR-613 is a novel approach for managing BC, which is worth further investigation.

Clinical Significance of Plasma Levels of Gluconeogenic Amino Acids in Esophageal Cancer Patients.

OBJECTIVE: Metabolic processes in the body of people with and without esophageal cancer (EC) are significantly different. Therefore, changes in the metabolism of amino acids in the body of EC patients can lead to metabolic disorders, such as increased gluconeogenesis. The aim of this study was the comparison of the plasma levels of gluconeogenic amino acids between patients with EC and the control group. METHODS: Plasma samples of 37 patients with EC who were selected before any treatment or surgery, and 37 healthy adults who did not have history of family cancer and malignant diseases were taken. Analysis of the plasma levels of amino acids including, alanine, asparagine, aspartate, glutamate, glutamine, glycine, serine, arginine, histidine, methionine, threonine, valine, tyrosine, isoleucine, phenylalanine, tryptophan was done by High Performance Liquid Chromatography (HPLC) based on reverse-phase-chromatography. Data analysis was done by SPSS-16 software. RESULTS: In the patient group the mean age ± SD was 63±13.64 and 21 (56.8%) were male.The plasma levels of the alanine, asparagine, histidine, methionine, threonine, valine amino acids in the patients with esophageal cancer was significantly reduced and glycine was increased (p-value<0.05). CONCLUSION: Gluconeogenic amino acids are the main precursor of glucose synthesis in the gluconeogenesis pathway. Cancer cells need more energy to grow and multiply, and glucose is used as the main fuel for cells. Given the importance of metabolic pathways in cancer cells, more detailed studies at the molecular level can provide new insights into early detection and appropriate treatment strategies for cancer.

Strontium and antimony serum levels in healthy individuals living in high- and low-risk areas of esophageal cancer.

BACKGROUND: It has been shown there is an upward trend for strontium (Sr) and antimony (Sb) levels from low-risk (LR) to high-risk (HR) areas of etiology of esophageal cancer in water, soil, and grains grown in Golestan province. In the present study, the serum levels of Sr and Sb were determined in healthy individuals living in these areas. METHODS: This cross-sectional study was performed on fasting blood serum of adult healthy individuals collected by cluster sampling. Subjects were divided into two groups, those living in either HR or LR areas. Strontium and antimony serum levels were measured using a graphite furnace atomic absorption spectroscopy. RESULTS: A total of 200 volunteers were enrolled from which 96 persons (48%) and 104 persons (52%) were from either HR or LR areas, respectively. The sex distribution was 40.9% male and 59.1% female, and the average age of enrolled people was 50.9 years. The average strontium levels were 30.44 ± 4.05 and 30.29 ± 3.74 µg/L in LR and HR, respectively. It also has been shown the average antimony levels were 15.21 ± 3.40, 14.81 ± 3.17, 15.13 ± 3.62, and 15.07 ± 3.62 µg/L in LR, HR, urban, and rural populations, respectively. CONCLUSION: The serum levels of strontium and antimony were not significantly different in healthy adults living in high- and low-risk areas of esophageal cancer. However, the average antimony serum levels in Golestan Province were above the reference interval in different countries.

microRNA-141 is associated with hepatic steatosis by downregulating the sirtuin1/AMP-activated protein kinase pathway in hepatocytes.

Sirtuin1 (SIRT1) is a crucial regulator of metabolism and it is implicated in the metabolic pathophysiology of several disorders inclusive of Type 2 diabetes and fatty liver disease (NAFLD). The aim of this study was to investigate the role of miR-141 in hepatic steatosis via regulation of SIRT1/AMP-activated protein kinase (AMPK) pathway in hepatocytes. Liver hepatocellular cells (HepG2) were treated with high concentration of glucose to be subsequently used for the assessment of miR-141 and SIRT1 levels in a model of hepatic steatosis. On the other hand, cells were transfected with miR-141 to investigate its effect on hepatocyte steatosis and viability as well as SIRT1 expression and activity along with AMPK phosphorylation. Targeting of SIRT1 by miR-141 was evaluated by bioinformatics tools and confirmed by luciferase reporter assay. Following the intracellular accumulation of lipids in HepG2 cells, the level of miR-141 was increased while SIRT1 mRNA and protein levels, as well as AMPK phosphorylation, was decreased. Transfection with miR-141 mimic significantly downregulated SIRT1 expression and activity while miR-141 inhibitor had the opposite effects. Additionally, modulation of miR-141 levels significantly influenced AMPK phosphorylation status. The results of luciferase reporter assay verified SIRT1 to be directly targeted by miR-141. miR-141 could effectively suppress SIRT1 and lead to decreased AMPK phosphorylation in HepG2 cells. Thus, miR-141/SIRT1/AMPK signaling pathway may be considered a potential target for the therapeutic management of NAFLD.

Serum levels of IL-32 in patients with type 2 diabetes mellitus and its relationship with TNF-α and IL-6.

Type 2 diabetes mellitus (T2DM) is an important public health worldwide. The main underlying mechanism of T2DM is insulin resistance which is associated with chronic inflammation. Interleukin-32 (IL-32) is a pro-inflammatory cytokine which has been implicated in pro-inflammatory responses of several human diseases. Previous studies have reported higher levels of IL-32 in inflammatory disease and obesity. The present study aimed to evaluate the serum concentrations of IL-32 in patients with T2DM and its association with cardio-metabolic parameters. This study was undertaken on 93 patients with TDM and 74 healthy controls. T2DM was diagnosed based on ADA criteria. Serum levels of IL-32, adiponectin, TNF-α, and IL-6 were measured by ELISA technique. Our findings revealed independent elevated levels of IL-32 in T2DM group (1061 (841.9-1601) pg/mL) compared to the control (630.4 (331.1-830.9) pg/mL). Furthermore, it was associated with increased risk of T2DM incidence. IL-32 indicated a positive correlation with body mass index, fasting blood glucose, TNF-α, and IL-6 in patients with T2DM. Furthermore, linear regression showed independent association between IL-32 and IL-6 plus TNF-α in patients' group. The results of the present study revealed higher levels of IL-32 in T2DM patients which have been associated with inflammatory markers. These results suggest the possible role of IL-32 in chronic inflammation in patients with T2DM.

Suppression of nicotinamide phosphoribosyltransferase expression by miR-154 reduces the viability of breast cancer cells and increases their susceptibility to doxorubicin.

BACKGROUND: Nicotinamide phosphoribosyltransferase (NAMPT) enzyme acts as the major enzyme in the nicotinamide adenine dinucleotide (NAD) synthesis salvage pathway. Deregulation of NAD could be associated with progression of several cancers such as breast cancer. Here, the consequence of NAMPT inhibition by miR-154 was investigated on breast cancer cells. METHODS: MDA-MB-231 and MCF-7 cancer cell lines were transfected with the mimic and inhibitors of miR-154-5p and their corresponding negative controls. Consequently, levels of NAMPT and NAD were assayed employing qRT-PCR, Western blotting and enzymatic method, respectively. Subsequently, flow cytometry and colorimetric methods were performed to evaluate apoptosis and cell viability. Bioinformatics analyses as well as luciferase assay were done to investigate whether the 3'-UTR of NAMPT is directly targeted by miR-154. RESULTS: According to the obtained results, NAMPT was recognized as a target for binding of miR-154 and the levels of this miRNA was inversely associated with both mRNA and protein levels of NAMPT in breast cancer cell lines. Functionally, miR-154 inhibited the NAD salvage pathway leading to a remarkable decrease in cell viability and increased rate of cell death. When breast cancer cells were simultaneously treated with doxorubicin and miR-154 mimic, cell viability was considerably reduced compared to treatment with doxorubicin alone in both cell lines. CONCLUSIONS: It was concluded that the inhibition of NAD production by miR-154 might be introduced as an appropriate therapeutic approach in order to improve breast cancer outcome either alone or in combination with other conventional chemotherapeutic agents.

MicroRNA-494 induces breast cancer cell apoptosis and reduces cell viability by inhibition of nicotinamide phosphoribosyltransferase expression and activity.

Breast cancer (BC) is the most prevalent cause of cancer-related death in women worldwide. BC is frequently associated with elevated levels of nicotinamide phosphoribosyltransferase (NAMPT) in blood and tumor tissue. MicroRNA-494 (miR-494) has been described to play key anti-tumor roles in human cancers. The aim of the present study was to investigate the inhibitory effect of miR-494 on NAMPT-mediated viability of BC cells. In this experimental study, MCF-7 and MDA-MB-231 cells were cultured and then transfected with miR-494 mimic, miR-494 inhibitor and their negative controls. The mRNA and protein expression of NAMPT were assessed using real-time PCR and Western blotting, respectively. Subsequently, intracellular NAD levels were determined by a colorimetric method. Finally, cell apoptosis was examined by flow cytometry. Bioinformatics evaluations predicted NAMPT as a miR-494 target gene which was confirmed by luciferase reporter assay. Our results showed an inverse relationship between the expression of miR-494 and NAMPT in both MCF-7 and MDA-MB-231 cell lines. miR-494 significantly down-regulated NAMPT mRNA and protein expression and was also able to reduce the cellular NAD content. Cell viability was decreased following miR-494 up-regulation. In addition, apoptosis was induced in MCF-7 and MDA-MB-231 cells by miR-494 mimic. Our findings indicate that miR-494 acts as a tumor suppressor and has an important effect in suppressing the growth of BC cells through NAMPT. Therefore, miR-494 might be considered as a novel therapeutic target for the management of human breast cancer.

Up-regulation of miR-381 inhibits NAD+ salvage pathway and promotes apoptosis in breast cancer cells.

Nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme involved in nicotinamide adenine dinucleotide (NAD) salvage pathway, is overexpressed in many human malignancies such as breast cancer. This enzyme plays a critical role in survival and growth of cancer cells. MicroRNAs (miRNAs) are among the most important regulators of gene expression, and serve as potential targets for diagnosis, prognosis, and therapy of breast cancer. Therefore, the aim of this study was to assess the effect of NAMPT inhibition by miR-381 on breast cancer cell survival. MCF-7 and MDA-MB-231 cancer cell lines were transfected with miR-381 mimic, inhibitor, and their corresponding negative controls (NCs). Subsequently, the level of NAMPT and NAD was assessed using real-time PCR, immuno-blotting, and enzymatic methods, respectively. In order to evaluate apoptosis, cells were labelled with Annexin V-FITC and propidium iodide and analyzed by flow cytometry. Bioinformatics analysis was performed to recognize whether NAMPT 3'-untranslated region (UTR) is a direct target of miR-381 and the results were authenticated by the luciferase reporter assay using a vector containing the 3'-UTR sequence of NAMPT. Our results revealed that the 3'-UTR of NAMPT was a direct target of miR-381 and its up-regulation decreased NAMPT gene and protein expression, leading to a notable reduction in intracellular NAD and subsequently cell survival and induction of apoptosis. It can be concluded that miR-381 has a vital role in tumor suppression by down-regulation of NAMPT, and it can be a promising candidate for breast cancer therapy.