Journal: Oxidative Medicine and Cellular Longevity

Article Title: Cysteine-Rich Whey Protein Isolate (CR-WPI) Ameliorates Erectile Dysfunction by Diminishing Oxidative Stress via DDAH/ADMA/NOS Pathway

doi: 10.1155/2022/8151917

Figure Lengend Snippet: Changes of the DDAH/NOS pathway and apoptosis-related proteins in penile tissues after 100 mg kg −1 of and 300 mg kg −1 CR-WPI supplementation for 60 days. (a) Representative immunofluorescence staining of the eNOS-positive penile specimen (red) in the sham, DM, 100 mg kg −1 CR-WPI, and 300 mg kg −1 CR-WPI groups for 60 days. (b) The effects of CR-WPI on the protein expression of DDAH, NOS, Caspase 3, and Caspase 9 in diabetic rats. β -Actin was used as a loading control. ∗ P < 0.01 and ∗∗ P < 0.01 indicate a significant difference compared with the DM group. # P < 0.05 and ## P < 0.01 indicate a significant difference compared with the sham group.

Article Snippet: The membrane was blocked with 5% skim milk for 1 h and subsequently incubated at 4°C overnight with primary antibodies against β -actin (1 : 1000; sc1616, Santa Cruz Biotechnology, Santa Cruz, CA, USA), α -SMA (1 : 1000; AF1032, Affinity Biosciences), eNOS (1 : 1000; AF0096, Affinity Biosciences), phosphorylated eNOS (Ser 1177) (p-eNOS; 1 : 500; sc81510, Santa Cruz Biotechnology, Santa Cruz, CA, USA), nNOS (1 : 600, SC5302, Santa Cruz Biotechnology, Santa Cruz, CA, USA), DDAH1 (1 : 1000; ab2231, Abcam, Cambridge, UK), DDAH2 (1 : 500; ab184166, Abcam, Cambridge, UK), Caspase 3 (D3R6Y) (1 : 1000; CST14220, Cell Signaling Technology), and Caspase 9 (1 : 2000; ab202068, Abcam, Cambridge, UK).

Techniques: Immunofluorescence, Staining, Expressing, Control

Journal: International journal of molecular medicine

Article Title: Naringenin modulates the NO‑cGMP‑PKG signaling pathway by binding to AKT to enhance osteogenic differentiation in hPDLSCs.

doi: 10.3892/ijmm.2024.5391

Figure Lengend Snippet: Figure 3. Ability of NAR to promote osteogenesis in human periodontal ligament stem cells is mitigated after L‑NAME (eNOS inhibitor) treatment. (A) Alkaline phosphatase staining in different treatment groups. (B) Alizarin red staining in different treatment groups. (C) NO levels in different treatment groups. (D) Gene expression levels of RUNX2, OPN and sGC in different treatment groups. (E‑G) Protein expression of RUNX2, OPN, sGC and p‑eNOS in various treatment groups. *P<0.05, **P<0.01, ***P<0.001. NAR, naringenin; eNOS, endothelial nitric oxide synthase; RUNX2, Runt‑related transcription factor; OPN, osteopontin; sGC, soluble guanylate cyclase; NO, nitric oxide; p‑, phosphorylated; t‑, total; L‑NAME, NG‑nitro‑L‑arginine methyl ester.

Article Snippet: Cells treated with NAR (10 μmol/l) were also treated with NG‐nitro‐L‐arginine methyl ester (L‐NAME; 70 μM) to inhibit endothelial nitric oxide synthase (eNOS), 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ; 20 μM) to inhibit soluble guanylate cyclase (sGC) and AKT inhibitor VIII (58 nM) to inhibit AKT, as recommended by the manufacturer (MedChemExpress for all).

Techniques: Staining, Gene Expression, Expressing

Journal: International journal of molecular medicine

Article Title: Naringenin modulates the NO‑cGMP‑PKG signaling pathway by binding to AKT to enhance osteogenic differentiation in hPDLSCs.

doi: 10.3892/ijmm.2024.5391

Figure Lengend Snippet: Figure 6. Impact of AKT inhibitor VIII on NAR‑mediated osteogenic promotion in human periodontal ligament stem cells. (A) Alkaline phosphatase staining in different treatment groups. (B) Alizarin red staining in different treatment groups. (C) Gene expression of RUNX2 and OPN in different treatment groups. (D‑F) Protein levels of RUNX2, OPN, p‑AKT and p‑eNOS in different treatment groups. *P<0.05, **P<0.01, ***P<0.001. NAR, naringenin; RUNX2, Runt‑related transcription factor; OPN, osteopontin; eNOS, endothelial nitric oxide synthase; p‑, phosphorylated; t‑, total.

Article Snippet: Cells treated with NAR (10 μmol/l) were also treated with NG‐nitro‐L‐arginine methyl ester (L‐NAME; 70 μM) to inhibit endothelial nitric oxide synthase (eNOS), 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ; 20 μM) to inhibit soluble guanylate cyclase (sGC) and AKT inhibitor VIII (58 nM) to inhibit AKT, as recommended by the manufacturer (MedChemExpress for all).

Techniques: Staining, Gene Expression

Journal: International journal of molecular medicine

Article Title: Naringenin modulates the NO‑cGMP‑PKG signaling pathway by binding to AKT to enhance osteogenic differentiation in hPDLSCs.

doi: 10.3892/ijmm.2024.5391

Figure Lengend Snippet: Figure 8. Schematic diagram of the mechanism by which naringenin promotes the osteogenic differentiation of human periodontal ligament stem cells. eNOS, endothelial nitric oxide synthase; p, phosphorylated; NO, nitric oxide; sGC, soluble guanylate cyclase; cGMP, cyclic guanosine monophosphate; TRPC6, transient receptor potential cation channel, subfamily C, member 6; PKG, protein kinase G.

Article Snippet: Cells treated with NAR (10 μmol/l) were also treated with NG‐nitro‐L‐arginine methyl ester (L‐NAME; 70 μM) to inhibit endothelial nitric oxide synthase (eNOS), 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ; 20 μM) to inhibit soluble guanylate cyclase (sGC) and AKT inhibitor VIII (58 nM) to inhibit AKT, as recommended by the manufacturer (MedChemExpress for all).

Techniques:

Journal: Veterinary Sciences

Article Title: Camel Whey Protein Attenuates Acute Heat Stress-Induced Kidney Injury in Rats by Up-Regulating CYP2J Activity and Activating PI3K/AKT/eNOS to Inhibit Oxidative Stress

doi: 10.3390/vetsci11110524

Figure Lengend Snippet: Protein expression of NGAL, KIM-1, and components of the PI3K/AKT/eNOS signaling pathway, indicators of renal injury under acute heat stress in CYP2J3 −/− rats. ( A ) shows the Western blotting electrophoresis of protein bands for NGAL, KIM-1, and the PI3K/AKT/eNOS signaling pathways in CYP2J3 −/− rats, with β-actin serving as an internal reference. ( B ) illustrates the relative expression of NGAL protein. ( C ) displays the relative expression of KIM-1 protein. ( D ) depicts the relative expression of PI3K protein. ( E ) presents the relative total expression of p-AKT protein, while ( F ) shows the relative total expression of p-eNOS protein. Data are expressed as mean ± SEM, based on six animals per group across the seven groups. “*” indicates a significant difference compared to the control group ( p < 0.05), and “**” indicates a highly significant difference compared to the control group ( p < 0.01); “#” indicates a significant difference compared with the HS group ( p < 0.05) and “##” indicates a highly significant difference compared with the HS group ( p < 0.01).

Article Snippet: Cytochrome P450 subenzyme CYP2J activity fluorescence quantitative detection kit (GENMED SCIENTIFICS INC., South San Francisco, CA, USA-GMS18190.2 v.A); Bovine whey protein (BWP) was purchased from Hilmar, TX, USA. β-actin antibody (ab8227) was purchased from Abcam; PI3K-P110 (AF5112), P-AKT (Ser473) (AF0016), AKT (AF6261), P-eNOS (Ser1177) (AF3247), and eNOS (AF0096) were purchased from Affinity Biosciences; sheep anti-rabbit IgG(H + L)-HRP (LK2001) was purchased from the Tianjin Three Arrows Biotechnology Co. (Tianjin, China).

Techniques: Expressing, Western Blot, Electrophoresis, Control