The effect of proteasome inhibitor (AM114) on apoptosis in IL-1β-treated peripheral blood macrophage cultured cells from rheumatoid arthritis patients

Selvarajan, Chitra; Ganesan, Nalini; Srinivasan, Lokeswari T.; Gopalakrishnan, Rajasekhar

doi:10.1016/j.injr.2015.10.005

Cited by 4 publications

(1 citation statement)

References 17 publications

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“…The IC50 concentrations of AM114 and Aspirin were 3.6 µmol and 3900 µmol in THP-1 cells, respectively. Previous reports have indicated that AM114 and Aspirin demonstrated cytotoxic effects on macrophages derived from the peripheral blood of rheumatoid arthritis (RA) patients at respective IC50 concentrations of 0.88 µmol and 1120 µmol [14]. Some of the studies explored the cytotoxicity of Aspirin against human adenocarcinoma HT-29 cell line and mouse neuro2a neuroblastoma cells with IC50 concentrations of 5 mmol and 2.5 mmol [15,5].…”

Section: Discussionmentioning

confidence: 99%

EXPLORING THE THERAPEUTIC POTENTIAL OF AM114: A BORONIC CHALCONE DERIVATIVE INDUCE APOPTOSIS AND SUPPRESS PROINFLAMMATORY CYTOKINES AND CHEMOKINES IN INTERLEUKIN-1β STIMULATED HUMAN THP-1 DERIVED MACROPHAGES

SELVARAJAN,

GANESAN

2024

Int J Pharm Pharm Sci

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Objective: Chalcones and their derivatives display a wide range of pharmacological activities. This study examined the effects of AM114, a boronic-chalcone derivative, on human THP-1-derived macrophages with and without interleukin-1β (IL-1β) stimulation. Methods: AM114 and Aspirin-treated THP-1-derived macrophages underwent activation with or without interleukin-1β. The IC50 concentrations of AM114 and Aspirin were determined through an MTT test. Apoptosis was measured using various techniques, including staining with acridine orange/Ethidium bromide, Hoechst 33342, and rhodamine 123 assays. Caspase-3 activity was measured using the spectrofluorimetric technique, while DNA fragmentation was assessed via agarose gel electrophoresis. Pro-inflammatory cytokines such as interleukin-6 (IL-6) and chemokines like interleukin-8 (IL-8) were measured using enzyme-linked immunosorbent assays. Results: AM114 and Aspirin showed dose-dependent cytotoxic effects on THP-1 macrophages. Induction of apoptosis was detected in AM114-treated THP-1 macrophages activated with IL-1β compared to macrophages without IL-1β. The gradation of dye uptake, membrane blebbing, increased caspase-3 activity, and DNA fragmentation ensures the induction of apoptosis, which indicates the cell's morphological changes, biochemical processes, and mitochondrial activity. Treating AM114 in IL-1β-activated THP-1 macrophages significantly reduced pro-inflammatory cytokines (IL-6) and chemokines (IL-8), suggesting its anti-cytokine potential in inflammatory diseases. Conclusion: The study results emphasize that AM114 could act as an anti-inflammatory agent by triggering apoptosis and reducing the release of cytokines and chemokines in inflammatory conditions. As a result, it may be used as a therapeutic option for inflammatory diseases.

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Section: Discussionmentioning

confidence: 99%

EXPLORING THE THERAPEUTIC POTENTIAL OF AM114: A BORONIC CHALCONE DERIVATIVE INDUCE APOPTOSIS AND SUPPRESS PROINFLAMMATORY CYTOKINES AND CHEMOKINES IN INTERLEUKIN-1β STIMULATED HUMAN THP-1 DERIVED MACROPHAGES

SELVARAJAN,

GANESAN

2024

Int J Pharm Pharm Sci

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Cysteine Cathepsins: In Health and Rheumatoid Arthritis

Ganesan¹

2017

Proteases in Human Diseases

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Effect of AM114 on apoptosis and proinflammatory cytokines in IL-1-beta treated human THP-1 derived macrophages

Selvarajan

Mohan²,

Ganesan

et al. 2023

Preprint

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Background: Chalcones and their derivatives are precursors of flavonoids and isoflavonoids abundantly present in edible plants. It displays a wide range of pharmacological activities, such as anticancer, antiinflammatory, antibacterial and antioxidant. AM-114 (3,5-Bis-[benzylidene-4-boronic acid]-1-methylpiperidin-4-one), a boronic-chalcone derivative exhibits potent anticancer activity through inhibition of the proteasome on human colon cancer cell line. However, the cytotoxic and anti-inflammatory effects of AM114 on THP-1 derived macrophages remain to be studied. Aim of the study: Inflammation is a complex reaction managed by a variety of immune cells such as monocytes and macrophages. In response to inflammatory stimuli, the macrophages secrete increased proinflammatory cytokines, chemokines such as interleukin-6 (IL-6) and interleukin-8 (IL-8). The proteasome complex is essential for several cellular processes including protein degradation, cellular differentiation and antigen presentation. In this study, human monocyte cell line THP-1 stimulated with interleukin-1-beta (IL-1-beta) was used as a model to investigate the in vitro effects of AM114, a proteasome inhibitor (PI), on apoptosis and release of proinflammatory cytokines. Materials and methods: The effect of AM114, with or without IL-1-beta stimulated THP-1derived macrophages, was assessed by comparing with aspirin. The cell viability was determined by MTT assay. The qualitative measurement of apoptosis was determined by acridine orange/ethidium bromide (AO/EtBr), Hoechst 33342 staining and rhodamine 123 assays. The quantitative measurement of apoptosis was carried out to determine the caspase-3 activity by spectrofluorimetric method and DNA fragmentation by agarose gel electrophoresis. The release of proinflammatory cytokines and chemokines such as IL-6 and IL-8 was also measured by enzyme linked immunosorbent assay (ELISA) method. The ANOVA was used to compare the different groups. Results: In the present study the IC50 concentration of AM114 and Aspirin on THP-1 cells were determined as 3.6 micromolar and 3900 micromolar respectively. The THP-1 derived macrophages treated with IL-1-beta showed insufficient apoptosis and increased release of proinflammatory cytokines. However the treatment of AM114, on stimulated THP-1 derived macrophages showed pronounced apoptotic features. The caspase-3 activity was increased 1.9 fold in AM114 with IL-1-beta treated THP-1 derived macrophages as compared to the unstimulated THP-1 derived macrophages pretreated with AM114. An increase of 3.2 fold in caspase-3 activity was observed in stimulated THP-1 derived macrophages pretreated with AM114 in comparison with stimulated THP-1 derived macrophages pretreated with aspirin. A significant increase of 5.0 and 3.0 fold DNA damage were observed in stimulated THP-1 derived macrophages treated with AM114 and aspirin as compared to unstimulated cells. Moreover AM114 treated cells showed significant decrease (p value less than 0.001) in the release of IL-6 and IL-8 on stimulated THP-1 derived macrophages. Conclusion: The results on the induction of apoptosis and suppression of proinflammatory cytokines suggest that AM114 is more effective than aspirin in stimulated THP-1 derived macrophages.

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The effect of proteasome inhibitor (AM114) on apoptosis in IL-1β-treated peripheral blood macrophage cultured cells from rheumatoid arthritis patients

Cited by 4 publications

References 17 publications

EXPLORING THE THERAPEUTIC POTENTIAL OF AM114: A BORONIC CHALCONE DERIVATIVE INDUCE APOPTOSIS AND SUPPRESS PROINFLAMMATORY CYTOKINES AND CHEMOKINES IN INTERLEUKIN-1β STIMULATED HUMAN THP-1 DERIVED MACROPHAGES

EXPLORING THE THERAPEUTIC POTENTIAL OF AM114: A BORONIC CHALCONE DERIVATIVE INDUCE APOPTOSIS AND SUPPRESS PROINFLAMMATORY CYTOKINES AND CHEMOKINES IN INTERLEUKIN-1β STIMULATED HUMAN THP-1 DERIVED MACROPHAGES

Cysteine Cathepsins: In Health and Rheumatoid Arthritis

Effect of AM114 on apoptosis and proinflammatory cytokines in IL-1-beta treated human THP-1 derived macrophages

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