Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 1. (a) Comparison of serum afamin content between control group and PCOS group. (b) The viability of KGN cells was decreased with increasing TP concentration. (c, d) Afamin inhibits the increase of MDA and 8-OHdG content in KGN cells caused by TP. (e-h) Afamin restored the decreased antioxidant enzyme activity including CAT, GR, GSH-Px, SOD in KGN cells caused by TP (i, j). Afamin reduces the increase in intracellular ROS levels caused by TP—Data represent mean ± standard error, Bar = 100 μm,*P < 0.05, **P <0.01.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques: Comparison, Control, Concentration Assay, Activity Assay

Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 2. (a) Afamin restored the decreased levels of mitochondrial membrane potential in KGN cells caused by TP. (b) Afamin reduces the increase in intracellular ROS levels caused by TP. (c) The relative mean MitoSOX fluorencence intensity was ana- lyzed by image J caused by TP. (d) Afamin restores the reduced ATP levels caused by TP. (e) Afamin restores the elevated activity of Capase3 caused by TP. (f) Afamin restored abnormal expression of apoptosis-related genes including Caspase 3, BAX, BCL2 caused by TP. (g) Representative cytometry data (dot-plot) for detection of apoptotic cells caused by TP (h) the average value of the percentage of apopotic cells annexin-positive cells for Q1-LR + Q1-UR in the population caused by TP. Data represent mean ± standard error, Bar = 100 μm,*P < 0.05, **P <0.01.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques: Membrane, Activity Assay, Expressing, Cytometry

Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 3. (a‒f) 3-AT (CAT inhibitor), MSA (GSH-PX inhibitor), BSO (GSH synthesis inhibitor) and BCNU (GR inhibitor) medi- ated the effects of afamin on MDA, 8-OHdG content and antioxidant enzyme activity including CAT, GR, GSH-Px, SOD in KGN cells caused by TP. (g, h) CAT (CAT inhibitor), MSA (GSH-PX inhibitor), BSO (GSH synthesis inhibitor) and BCNU (GR inhibitor) mediated the effects of afamin on mitochondrial membrane potential. Data represent mean ± standard error, Bar = 100 μm,*P < 0.05, **P < 0.01.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques: Activity Assay, Membrane

Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 4. (a‒f) Mitochondrial respiratory chain complex I inhibitor (ROT) mediated the effects of afamin on MDA, 8-OHdG con- tent and antioxidant enzyme activity including CAT, GR, GSH-Px, SOD in KGN cells caused by TP. (g) Effects of Afamin on the activities of mitochondrial respiratory chain complex I in KGN cells exposed to TP. (h) After treatment with ROT and addi- tion of afamin, TMRM was analyzed by cellular imaging caused by TP. Data represent mean±standard error, Bar = 100 μm,*P < 0.05, **P < 0.01.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques: Activity Assay, Imaging

Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 5. (a) After introducing the SIRT1 siRNA into KGN cells, effects of SiSIRT1-1, SiSIRT1-2, SiSIRT1-3 and SiSIRT1-4 on SIRT1 gene expression was detected. (b) After introducing the SIRT1 siRNA into KGN cells, effects of SiSIRT1-1 and SiSIRT1- 2 on SIRT1 protein expression was detected. (c) When exposed to TP, the effects of afamin on SIRT1 the protein expression was detected. (d) When exposed to TP, the effects of afamin on SIRT3 and SIRT5 the protein expression was detected. (e) When exposed to TP, the effects of afamin on HO-1 the protein expression was detected. (f‒i) When silencing of SIRT1, the effects of afamin on MDA content and antioxidant enzyme activity including GR, GSH-Px, SOD in KGN cells were detected caused by TP. (j) When silencing of SIRT1, the effects of afamin on ATP was analyzed by according to corresponding assay kit caused by TP. (k) When silencing of SIRT1, MitoSOX fluorencence intensity was analyzed by Image J caused by TP and afamin. (l) When silencing of SIRT1, MitoSOX fluorencence was detected by cellular imaging caused by TP and afamin. Data represent mean ± standard error, Bar = 100 μm,*P < 0.05, **P < 0.01.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques: Gene Expression, Expressing, Activity Assay, Imaging

Journal: Molecular Biology

Article Title: Afamin Ameliorates Testosterone Propionate (TP)-Induced Oxidative Stress and Mitochondrial Damage in Human Ovarian Granulosa Tumor Cells (KGN) by Upregulating the Expression of SIRT1

doi: 10.1134/s0026893324060074

Figure Lengend Snippet: Fig. 8. Schematic diagram summarizes the protective effects of afamin ameliorates TP-induced Oxidative Stress and mitochon- dria damage of KGN cells in a SIRT1-dependent manner. Afamin might ameliorate TP-induced oxidative stress of KGN cells by improving the antioxidant capacity of SOD, CAT, GSH-Px, and GSH and restoring the mitochondrial function. Meanwhile, afamin ameliorated oxidative stress-induced KGN cell apoptosis via Caspase3, BAX, and BCL2.

Article Snippet: Also, KGN cells were treated with afamin (50 ng/mL) and then exposed to TP (50 μΜ) for 12 h in the absence or presence of the mitochondrial respiratory chain complex I inhibitor rotenone (ROT; Med Chem Express (MCE) Inc., Monmouth Junction, NJ, USA), at 2 μΜ.

Techniques:

Journal: Frontiers in Immunology

Article Title: Inhibition of EETosis with an anti-citrullinated histone antibody: a novel therapeutic approach for eosinophilic inflammatory disorders

doi: 10.3389/fimmu.2025.1533407

Figure Lengend Snippet: Histone citrullination is required for EETosis. (A) Citrullinated nucleosome detection in EETs harvested 3 hours post stimulation with 2 µM A23187, 40.5 nM PMA, and 0.5 µM platelet activating factor (PAF) without or with 1 U/mL thrombin ( n = 2-4). Quantification of A23187- and PMA-induced EET release at t = 3 h in the presence of different concentrations of JBI-589 (B) and AFM-30a (C) ( n = 4-7). Data were normalized to A23187- or PMA-induced EET release without inhibitor (set as 100% EET release). **** P <0.0001, ordinary one-way ANOVA with Dunnett’s multiple comparison test.

Article Snippet: To study EETosis signaling pathways, eosinophils were pre-incubated for 30 min at 37°C with indicated concentration of PAD2 inhibitor AFM-30a hydrochloride (MedChemExpress, HY-125099A) or PAD4 inhibitors JBI-589 (MedChemExpress, HY-153450), GSK484 (MedChemExpress, HY-100514), or BMS-P5 (MedChemExpress, 33581) before addition of the stimulus.

Techniques: Comparison