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BMC Microbiology Apr 2024Leishmaniasis as a neglected tropical disease (NTD) is caused by the inoculation of Leishmania parasites via the bite of phlebotomine sand flies. After an infected bite,...
BACKGROUND
Leishmaniasis as a neglected tropical disease (NTD) is caused by the inoculation of Leishmania parasites via the bite of phlebotomine sand flies. After an infected bite, a series of innate and adaptive immune responses occurs, among which neutrophils can be mentioned as the initiators. Among the multiple functions of these fighting cells, neutrophil extracellular traps (NETs) were studied in the presence of Leishmania major promastigotes and salivary gland homogenates (SGH) of Phlebotomus papatasi alone, and in combination to mimic natural conditions of transmission.
MATERIAL & METHODS
The effect of L. major and SGH on NETs formation was studied in three different groups: neutrophils + SGH (NS), neutrophils + L. major (NL), neutrophils + L. major + SGH (NLS) along with negative and positive controls in 2, 4 and 6 h post-incubation. Different microscopic methods were used to visualize NETs comprising: fluorescence microscopy by Acridine Orange/ Ethidium Bromide staining, optical microscopy by Giemsa staining and scanning electron microscopy. In addition, the expression level of three different genes NE, MPO and MMP9 was evaluated by Real-Time PCR.
RESULTS
All three microscopical methods revealed similar results, as in NS group, chromatin extrusion as a sign of NETosis, was not very evident in each three time points; but, in NL and especially NLS group, more NETosis was observed and the interaction between neutrophils and promastigotes in NL and also with saliva in NLS group, gradually increased over times. Real-time reveals that, the expression of MPO, NE and MMP9 genes increased during 2 and 4 h after exposure, and then decreased at 6 h in most groups.
CONCLUSION
Hence, it was determined that the simultaneous presence of parasite and saliva in NLS group has a greater impact on the formation of NETs compared to NL and NS groups.
Topics: Animals; Humans; Phlebotomus; Leishmania major; Matrix Metalloproteinase 9; Extracellular Traps; Neutrophils; Salivary Glands
PubMed: 38575882
DOI: 10.1186/s12866-024-03270-z -
Scientific Reports Aug 2023The implication of inflammation in the pathophysiology of several types of cancers has been under intense investigation. Conjugated fatty acids can modulate inflammation...
The implication of inflammation in the pathophysiology of several types of cancers has been under intense investigation. Conjugated fatty acids can modulate inflammation and present anticancer effects, promoting cancer cell death. In this paper, we evaluated the efficacy of new conjugated fatty acids isolated from marine Opisthopterus tardoore (Tapra fish) in human breast cancer cell lines MCF-7. Linoelaidic acid, a marine fish (O. tardoore) derived unsaturated fatty acids, showed effective anticancer activity against MCF-7. Cell viability (MTT) assay revealed a dose-dependent decline in cancer cell viability. It was noteworthy that 5 µM linoelaidic acid decreased the MCF-7 cell viability by 81.82%. Besides that, linoelaidic acid significantly (P< 0.05) increased the level of tumor necrosis factor-α (TNF-α) and interleukin-1 receptor antagonist (IL-1ra) studied by ELISA. Not only that, linoelaidic acid significantly decreased the reduced glutathione level and increased the oxidized glutathione level in MCF-7 cells indicating the oxidative stress inside the cell. Two different cell staining methods with acridine orange-ethidium bromide and DAPI confirmed that the linoelaidic acid rendered their detrimental effect on cancer cells. To decipher the mode of apoptosis Western blotting was performed in which the expression pattern of several proteins (p53, IL-10, and IL-1ra) established the apoptosis in the studied cell lines after linoelaidic acid exposure. Hence it may be conferred that linoelaidic acid has prompt anticancer activity. Therefore this drug can be used further for the treatment of cancer.
Topics: Humans; Linoleic Acid; MCF-7 Cells; Reactive Oxygen Species; Interleukin 1 Receptor Antagonist Protein; Cell Death; Fatty Acids; Dietary Fats, Unsaturated; Caspases
PubMed: 37644076
DOI: 10.1038/s41598-023-34885-3 -
International Journal of Pharmaceutics Apr 2024Gold core mesoporous silica shell (AuMSS) nanorods are multifunctional nanomedicines that can act simultaneously as photothermal, drug delivery, and bioimaging agents....
Gold core mesoporous silica shell (AuMSS) nanorods are multifunctional nanomedicines that can act simultaneously as photothermal, drug delivery, and bioimaging agents. Nevertheless, it is reported that once administrated, nanoparticles can be coated with blood proteins, forming a protein corona, that directly impacts on nanomedicines' circulation time, biodistribution, and therapeutic performance. Therefore, it become crucial to develop novel alternatives to improve nanoparticles' half-life in the bloodstream. In this work, Polyethylenimine (PEI) and Red blood cells (RBC)-derived membranes were combined for the first time to functionalize AuMSS nanorods and simultaneously load acridine orange (AO). The obtained results revealed that the RBC-derived membranes promoted the neutralization of the AuMSS' surface charge and consequently improved the colloidal stability and biocompatibility of the nanocarriers. Indeed, the in vitro data revealed that PEI/RBC-derived membranes' functionalization also improved the nanoparticles' cellular internalization and was capable of mitigating the hemolytic effects of AuMSS and AuMSS/PEI nanorods. In turn, the combinatorial chemo-photothermal therapy mediated by AuMSS/PEI/RBC_AO nanorods was able to completely eliminate HeLa cells, contrasting with the less efficient standalone therapies. Such data reinforce the potential of AuMSS nanomaterials to act simultaneously as photothermal and chemotherapeutic agents.
Topics: Humans; HeLa Cells; Photothermal Therapy; Erythrocyte Membrane; Silicon Dioxide; Gold; Tissue Distribution; Phototherapy; Antineoplastic Agents; Nanotubes; Doxorubicin; Neoplasms
PubMed: 38493844
DOI: 10.1016/j.ijpharm.2024.124007 -
Food Science & Nutrition Mar 2024Influenza remains one of the most serious infectious diseases. Gallic acid is one of the most common and representative phenolic acids found in various plants. This is...
Influenza remains one of the most serious infectious diseases. Gallic acid is one of the most common and representative phenolic acids found in various plants. This is an interesting subject to explore how gallic acid could inhibit H1N1 influenza virus infection by reducing the production of virulent proteins and interrupting autophagy machinery for influenza virus replication on the host cell. Cellular viability was assessed by XTT assay. The inhibitory effects on the H1N1 influenza virus were assessed by hemagglutination assay, plaque assay, and qRT-PCR. Western blot analysis was used for detecting protein levels of M1, M2, NP, LC3B, and beclin-1. Autophagy activity was demonstrated by acridine orange staining assay. The result demonstrated that there was no cytotoxic effect of gallic acid on A549 cells, and gallic acid could restore the cellular viability of H1N1 influenza virus-infected A549 cells within the experimental concentration treatment. Moreover, gallic acid could effectively restrain viral activity of the H1N1 influenza virus. After the treatment of gallic acid, the production of virulent H1N1 influenza virus proteins, that is, M1, M2, and NP protein were reduced. As for autophagic mechanism, both of the LC3B II conversion and the level ratio of LC3B II to LC3B I were notably decreased. The acridine orange staining assay also revealed decreased accumulation of autophagosomes in H1N1 influenza virus-infected cells. In conclusion, gallic acid suppresses H1N1 influenza viral infectivity through restoration of autophagy pathway and inhibition of virulent M1, M2, and NP protein production.
PubMed: 38455214
DOI: 10.1002/fsn3.3852 -
Saudi Pharmaceutical Journal : SPJ :... Mar 2024is traditionally used to treat breast cancer in several Arab countries. Scientific studies have reported different effects of this plant on some cancer cell lines. The...
is traditionally used to treat breast cancer in several Arab countries. Scientific studies have reported different effects of this plant on some cancer cell lines. The current study determined the anti-cancer potential of the methanolic extract of against four different types of breast cancer cell lines . The extract was prepared by maceration and phytoconstituents were identified by LC-MS analysis. The IC value was determined against MDA-MB-231, MCF-7, 4 T1, and MCF-10 cell lines using the MTT assay. Further investigations were carried out using IC concentration of the extract (40.09 µg/ml) to determine live/dead cells by acridine orange/ethidium bromide staining. The effect on the expression of reactive oxygen species (ROS) was evaluated by flow cytometry. The results were analyzed using one-way ANOVA followed by Tukey's test. The LC-MS analysis revealed the presence of 34 and 30 phytoconstituents in positive and negative modes respectively. The extract was most effective against 4 T1 cells in a dose-dependent manner (P < 0.001) with an IC value of 40.09 µg/ml and showed negligible effect against MCF-10 cells. It increased apoptosis in 77.84 % of 4 T1 cells, as determined by acridine orange/ethidium bromide staining. The extract also increased the ROS expression in the 39.57 % of 4 T1 cells. The study results showed that Ephedra foeminea extract possesses an anti-cancer effect against 4 T1 cells by increasing the expression of ROS and inducing apoptosis in the 4 T1 cells. The result suggests methanolic extract possesses a reasonable anti-cancer effect due to its effect on apoptosis and oxidative pathways. The results confirm the traditional belief that is effective against breast cancerز.
PubMed: 38328794
DOI: 10.1016/j.jsps.2024.101960 -
Advances in Clinical and Experimental... Feb 2024Asthma is a chronic illness that causes recurrent inflammation and airway constriction. The primary risk factors for asthma development are exposure to environmental...
BACKGROUND
Asthma is a chronic illness that causes recurrent inflammation and airway constriction. The primary risk factors for asthma development are exposure to environmental allergens and house dust mites, which can trigger deoxyribonucleic acid (DNA) damage. Oxidative stress can also cause DNA impairments and plays a crucial role in the progression of human immunological disorders.
OBJECTIVES
The aim of the study was to evaluate the effects of oridonin (ORD) on proliferation, inflammation and apoptosis in interleukin 4 (IL-4)-stimulated human bronchial epithelial (16HBE) cells.
MATERIAL AND METHODS
Proliferation was assessed using a 5-Bromo-2-deoxyuridine (BrdU) assay, while acridine orange (AO), ethidium bromide (EB), propidium iodide, and 4',6-diamidino-2-phenylindole (DAPI) measured apoptosis. The protein expression levels of apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), cleaved caspase-1, and nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) were detected with western blot.
RESULTS
The results established that IL-4 stimulation markedly decreased (p < 0.05) the proliferation of 16HBE cells, while the administration of ORD increased their proliferation. Apoptosis and DNA damage were enhanced in the IL-4-stimulated group, whereas ORD exhibited anti-apoptotic activity. Moreover, the treatment with ORD significantly reduced (p < 0.05) the IL-4-induced expression of cleaved caspase-1, ASC and NLRP3 proteins.
CONCLUSIONS
The findings suggest that NLRP3 is a direct target for ORD-mediated anti-inflammatory actions in injured 16HBE cells. Therefore, ORD may be a novel therapy against NLRP3-related disorders, including pediatric asthma (PA).
Topics: Child; Humans; NLR Family, Pyrin Domain-Containing 3 Protein; Inflammasomes; Interleukin-4; Apoptosis; Inflammation; Caspase 1; Epithelial Cells; Asthma; DNA; Interleukin-1beta; Diterpenes, Kaurane
PubMed: 37486694
DOI: 10.17219/acem/166253 -
Balkan Medical Journal Jan 2024The glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide (LIRA) is a potential hypoglycemic drug with anti-atherosclerosis (AS) effects. Autophagy in the...
BACKGROUND
The glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide (LIRA) is a potential hypoglycemic drug with anti-atherosclerosis (AS) effects. Autophagy in the vascular smooth muscle cells (VSMCs) facilitates AS. However, the role of autophagy in the anti-AS mechanism of LIRA remains unclear.
AIMS
To examine the role and mechanisms of autophagy in LIRA’s improvement of the biological characteristics of VSMCs in high glucose conditions.
STUDY DESIGN
Experimental animal study.
METHODS
VSMCs isolated from the thoracic aorta of male SD rats were subjected to a high glucose (HG) condition (25 mM) in Dulbecco’s Modified Eagle’s Medium with or without LIRA, the GLP-1 receptor antagonist exendin9-39 (Exe9-39), a phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), and autophagy inhibitors (3-methyladenine [3-MA] and bafilomycin A1 [Baf A1]). Acridine orange staining, western blotting, transmission electron microscopy, and mCherry-GFP-LC3 transfection were performed to evaluate the autophagy flux. Additionally, VSMC migration, calcification, proliferation, and apoptosis in HG conditions were observed.
RESULTS
Addition of LIRA alone or in combination with autophagy inhibitors significantly downregulated Beclin, increased the LC3-II/LC3-I ratio, and upregulated p62 in VSMCs in HG conditions. Furthermore, autophagolysosome formation was markedly curbed after treatment with LIRA and/or autophagy inhibitors. Inhibition of autophagy by LIRA and/or the autophagy inhibitors attenuated VSMC phenotype conversion, proliferation, migration, and calcification and promoted VSMC apoptosis in HG conditions. This protective role of LIRA was augmented by LY294002, but inhibited by Exe9-39.
CONCLUSION
LIRA plays a significant role in the improvement of the biological features of VSMCs in HG conditions.
Topics: Rats; Male; Animals; Liraglutide; Muscle, Smooth, Vascular; Signal Transduction; Phosphatidylinositol 3-Kinases; Rats, Sprague-Dawley; Autophagy; Atherosclerosis; Glucose
PubMed: 37953594
DOI: 10.4274/balkanmedj.galenos.2023.2023-8-44 -
PloS One 2024N-linked glycosylation is a pivotal post-translational modification that significantly influences various aspects of protein biology. Autophagy, a critical cellular...
N-linked glycosylation is a pivotal post-translational modification that significantly influences various aspects of protein biology. Autophagy, a critical cellular process, is instrumental in cell survival and maintenance. The hepatitis B virus (HBV) has evolved mechanisms to manipulate this process to ensure its survival within host cells. Significantly, post-translational N-linked glycosylation in the large surface protein of HBV (LHBs) influences virion assembly, infectivity, and immune evasion. This study investigated the role of N-linked glycosylation of LHBs in autophagy, and its subsequent effects on HBV replication and secretion. LHBs plasmids were constructed by incorporating single-, double-, and triple-mutated N-linked glycosylation sites through amino acid substitutions at N4, N112, and N309. In comparison to the wild-type LHBs, N-glycan mutants, including N309Q, N4-309Q, N112-309Q, and N4-112-309Q, induced autophagy gene expression and led to autophagosome accumulation in hepatoma cells. Acridine orange staining of cells expressing LHBs mutations revealed impaired lysosomal acidification, suggesting potential blockage of autophagic flux at later stages. Furthermore, N-glycan mutants increased the mRNA expression of HBV surface antigen (HBsAg). Notably, N309Q significantly elevated HBx oncogene level. The LHBs mutants, particularly N309Q and N112-309Q, significantly enhanced HBV replication, whereas N309Q, N4-309Q, and N4-112-309Q markedly increased HBV progeny secretion. Remarkably, our findings demonstrated that autophagy is indispensable for the impact of N-linked glycosylation mutations in LHBs on HBV secretion, as evidenced by experiments with a 3-methyladenine (3-MA) inhibitor. Our study provides pioneering insights into the interplay between N-linked glycosylation mutations in LHBs, host autophagy, and the HBV life cycle. Additionally, we offer a new clue for further investigation into carcinogenesis of hepatocellular carcinoma (HCC). These findings underscore the potential of targeting either N-linked glycosylation modifications or the autophagic pathway for the development of innovative therapies against HBV and/or HCC.
Topics: Humans; Hepatitis B virus; Carcinoma, Hepatocellular; Glycosylation; Liver Neoplasms; Hepatitis B Surface Antigens; Hepatitis B; Autophagy; Membrane Proteins; Polysaccharides
PubMed: 38489292
DOI: 10.1371/journal.pone.0299403 -
BMC Complementary Medicine and Therapies Jun 2024Cervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS)...
BACKGROUND
Cervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation.
METHODS
The proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS.
RESULTS
EESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes.
CONCLUSION
This study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer.
Topics: Humans; Sophora; HeLa Cells; Uterine Cervical Neoplasms; Female; Cell Proliferation; Plant Extracts; Apoptosis; Antineoplastic Agents, Phytogenic; Mice; Ethanol
PubMed: 38831394
DOI: 10.1186/s12906-024-04502-5