sod1 inhibitor  (MedChemExpress)

Journal: Stem Cell Research & Therapy

Article Title: hucMSC-derived exosomes targeting macrophage polarization attenuate systemic inflammation in T1DM via INS/SOD1 delivery

doi: 10.1186/s13287-025-04521-0

Figure Lengend Snippet: Anti-inflammatory protein components in hucMSC-EXOs. ( A-F ) qPCR was performed to compare the effects of heat-inactivated hucMSC-EXOs and untreated hucMSC-EXOs on the expression levels of inflammatory cytokines in macrophages. ( G ) Topological analysis of hucMSC-EXOs proteins (top 200 by IBAQ value) via the STRING database, identifying the top 40 hub proteins ranked by interaction degree. Their degrees of interaction are visualized as a concentric circle plot. ( H-K ) ELISA was used to detect the INS ( H-I ) content and SOD1 ( J-K ) content of the hucMSCs culture supernatant and hucMSC-EXOs. (L‒O) qPCR analysis the effects of EXO alone, EXO combined with S961, and EXO combined with ATN-224 on the expression of inflammatory factors in macrophages. ( P ) WB experiment analysis the influence of EXO alone, EXO combined with S961, and EXO combined with ATN-224 treatment on PI3K/AKT and P65 phosphorylation in macrophages. * P < 0.05; ** P < 0.01 ; *** P < 0.001

Article Snippet: THP-1 cells were differentiated into M0 macrophages with 100 nM phorbol myristate acetate (PMA; HY-18739, MCE, US) for 24 h, and then divided into six experimental groups: PMA control, lipopolysaccharide (LPS; 50 ng/mL; L2880, Sigma, US), LPS + 5 nM INS (GC2646, Genxion, China), LPS + 1 μg SOD1 (HY-P71048A, MCE, US), LPS + hucMSC-EXOs (8.5 × 10^5 particles), LPS + heat-inactivated hucMSC-EXOs (8.5 × 10^5 particles), LPS + hucMSC-EXOs + 5 nM INS receptor antagonist S961 (HY-P2093B, MCE, US), and LPS + hucMSC-EXOs + 20 nM SOD1 inhibitor ATN-224 (HY-16074, MCE, US).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Phospho-proteomics

Journal: Microbiology Spectrum

Article Title: Superoxide Initiates the Hyphal Differentiation to Microsclerotia Formation of Macrophomina phaseolina

doi: 10.1128/spectrum.02084-21

Figure Lengend Snippet: Microsclerotia stimulation assay supported that O 2 − stimulates microsclerotia formation in M. phaseolina . (A) Mycelial growth rate at different concentrations of H 2 O 2 -amended PDA plates. (B) Mycelial growth rate at different concentrations of diethyldithiocarbamate trihydrate (DETC)-amended PDA plates. The best concentration was defined as the highest concentration not affecting mycelial growth, as indicated by the red line, which was 0.3 mM for DETC and 1 mM for H 2 O 2 . (C) Microsclerotia stimulation rate of 0.3 mM DETC and 1 mM H 2 O 2 . An increase of microsclerotia formation of approximately 10% was estimated in 0.3 mM DETC-amended PDA plates. (D) Microsclerotia formation in a plate and in the area 3.5 cm from the center of a 9-cm petri dish. The DETC treatment stimulated high microsclerotia formation. Three biological replicates were included in each experimental repeat, and the experiment was repeated three times. The bars indicate 1 mm. (E) 5-mm plugs were sampled at the 3.5 cm from the center of a 9-cm petri dish for photographing before and after staining. The bars indicate 1 mm. Each staining was repeated three times, and each included three independent biological replicates. Because DETC blocks the process of O 2 − into H 2 O 2 without reducing microsclerotia formation, the results support that O 2 − is the main ROS stimulus for microsclerotia formation in M. phaseolina .

Article Snippet: The SOD1 inhibitor sodium diethyldithiocarbamate trihydrate (DETC) (Alfa Aesar, Thermo Fisher Scientific, Ward Hill, MA), and 30% vol H 2 O 2 (PanReac AppliChem) were freshly prepared in the 9-cm petri dish PDA plates to minimize the reduction of oxidants, and the assay was conducted as described for the antioxidants.

Techniques: Concentration Assay, Staining

Journal: Molecular Therapy Oncolytics

Article Title: Oleanolic acid blocks the purine salvage pathway for cancer therapy by inactivating SOD1 and stimulating lysosomal proteolysis

doi: 10.1016/j.omto.2021.08.013

Figure Lengend Snippet: OA selectively inactivates SOD1 to degrade HGPRT and 5′-NT via the ROS/AMPK/mTORC1/macroautophagy/lysosome pathway (A) Western blot showing the time course of the effects of OA treatment (200 μM) on the abundance of phospho-AMPK (Thr172), AMPK, phospho-ACC (Ser79), ACC, phospho-ULK1 (Ser555), phospho-ULK1 (Ser757), ULK1, phospho-4EBP1 (Thr37/46), 4EBP1, and LC3-I/II in A549 cells. (B) The effects of two ULK1 inhibitors, ULK-101 (1 μM) and MRT68921 (1 μM), on levels of phospho-ATG14 (Ser29), ATG14, LC3-I/II, HGPRT, and 5′-NT in OA-treated A549 cells. (C) Western blot showing the influence of 3-MA (1 mM) on HGPRT, 5′-NT, and LC3-I/II expression in OA-treated A549 cells. (D) Western blot showing the time course of the effects of 200 μM OA treatment on the expression of phospho-AMPK (Thr172), AMPK, phospho-ACC (Ser79), ACC, phospho-ULK1 (Ser757), ULK1, phospho-4EBP1 (Thr37/46), 4EBP1, LC3-I/II, HGPRT, and 5′-NT in A549 cells with and without PRKAA1 KO. (E) Measurement of reactive oxygen species (ROS) generation induced by OA (200 μM) and ROS removal following N -acetyl- l -cysteine (NAC) (5 mM) treatment in A549 cells. (F) Time course of the reversion of phospho-AMPK (Thr172), phospho-ACC (Ser79), ACC, HGPRT, 5′-NT, and LC3-I/II in OA-treated A549 cells after NAC (5 mM). The values of phospho-AMPK (Thr172) were normalized as follows: the raw abundance of phospho-AMPK (Thr172) of each lane was first normalized by the corresponding actin, and the actin-normalized proteins of all lanes were then further normalized by the actin-normalized protein on the fourth lane (control group treated by OA for 8 h). Normalized phospho-ACC (Ser79) was acquired by the same computation approach. (G) Restoration of impaired cell proliferation and PCNA expression by NAC with OA treatment (200 μM). Viable cell proliferation was measured with an ATPlite kit. (H) Measurement of ROS generation in OA-treated A549 cells with and without SOD1 overexpression. (I) Under OA treatment, SOD1 overexpression enhanced HGPRT and 5′-NT stability and restored PCNA expression. Error bars represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, Student’s t test. Ns, no significance.

Article Snippet: The reagents used in this study included OA (Sigma-Aldrich, St. Louis, MO, USA), dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA), hypoxanthine (MedChemExpress, Monmouth Junction, NJ, USA), inosine (MedChemExpress, Monmouth Junction, NJ, USA), cycloheximide (Sigma-Aldrich, St. Louis, MO, USA), MG132 (MedChemExpress, Monmouth Junction, NJ, USA), CQ (Cell Signaling Technology, Boston, MA, USA), BAF (Cell Signaling Technology, Boston, MA, USA), ULK-101 (MedChemExpress, Monmouth Junction, NJ, USA), MRT68921 (MedChemExpress, Monmouth Junction, NJ, USA), IMP (MedChemExpress, Monmouth Junction, NJ, USA), 3-MA (MedChemExpress, Monmouth Junction, NJ, USA), NAC (MedChemExpress, Monmouth Junction, NJ, USA), SOD1 inhibitor LCS-1 (Sigma-Aldrich, St. Louis, MO, USA), and NP40 buffer (Beyotime, Shanghai, China).

Techniques: Western Blot, Expressing, Over Expression

Journal: Molecular Therapy Oncolytics

Article Title: Oleanolic acid blocks the purine salvage pathway for cancer therapy by inactivating SOD1 and stimulating lysosomal proteolysis

doi: 10.1016/j.omto.2021.08.013

Figure Lengend Snippet: The PSP is overactivated in human lung cancer and breast cancer, with a negative linkage to patient survival (A) Western blot showing HGPRT and 5′-NT expression in tumorous and paired normal adjacent tissues from patients with lung adenocarcinoma (n = 5). (B) Representative immunohistochemistry stained images of lung tissue microarrays using HGPRT and 5′-NT antibodies from samples obtained from patients with lung cancer (n = 34). Scale bars for ×40 images, 625 μm; scale bars for ×400 images, 50 μm. The midlines of the boxplots represent the median values of the data, with the upper and lower limits of the box indicating the third and first quartiles and the whiskers of the boxplot up to 1.5 times the interquartile ranges. The p values were calculated with non-parametric Wilcoxon rank-sum tests. N, paired normal adjacent lung tissues; T, tumorous lung tissues. (C) Representative immunohistochemistry stained images of lung tissue microarrays using HGPRT and 5′-NT antibodies in samples from patients with breast cancer (n = 20). Scale bars for ×40 images, 625 μm; scale bars for ×400 images, 50 μm. The midlines of the boxplots represent the median value of the data, with the upper and lower limits of the box indicating the third and first quartiles and the whiskers up to 1.5 times the interquartile ranges. The p values were calculated with non-parametric Wilcoxon rank-sum tests. N, paired normal adjacent breast tissues; T, tumorous breast tissues. (D) Enzyme activity of HGPRT and 5′-NT in tumorous and paired normal adjacent tissues derived from patients with lung adenocarcinoma (n = 5). The p values were acquired by Student’s t test. (E) Differentially expressed PSP metabolites between tumorous and normal adjacent lung tissues from patients with lung cancer (n = 34). (F and G) Kaplan-Meier curves showing the association of expression of HPRT1 or NT5E and overall survival of patients with lung or breast cancer from the public TCGA and GTEx databases. The p values were gained by log-rank test. (H) A model proposed by the current study for the herbal compound OA. OA impaired cancer cell growth by blocking the PSP by degrading HGPRT and 5′-NT in this pathway through the SOD1/ROS/AMPK/mTORC1/autophagy/lysosome pathway.

Article Snippet: The reagents used in this study included OA (Sigma-Aldrich, St. Louis, MO, USA), dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA), hypoxanthine (MedChemExpress, Monmouth Junction, NJ, USA), inosine (MedChemExpress, Monmouth Junction, NJ, USA), cycloheximide (Sigma-Aldrich, St. Louis, MO, USA), MG132 (MedChemExpress, Monmouth Junction, NJ, USA), CQ (Cell Signaling Technology, Boston, MA, USA), BAF (Cell Signaling Technology, Boston, MA, USA), ULK-101 (MedChemExpress, Monmouth Junction, NJ, USA), MRT68921 (MedChemExpress, Monmouth Junction, NJ, USA), IMP (MedChemExpress, Monmouth Junction, NJ, USA), 3-MA (MedChemExpress, Monmouth Junction, NJ, USA), NAC (MedChemExpress, Monmouth Junction, NJ, USA), SOD1 inhibitor LCS-1 (Sigma-Aldrich, St. Louis, MO, USA), and NP40 buffer (Beyotime, Shanghai, China).

Techniques: Western Blot, Expressing, Immunohistochemistry, Staining, Activity Assay, Derivative Assay, Blocking Assay