Journal: Oxidative Medicine and Cellular Longevity

Article Title: Catalpol Enhances Random-Pattern Skin Flap Survival by Activating SIRT1-Mediated Enhancement of Autophagy

doi: 10.1155/2022/5668226

Figure Lengend Snippet: CAT alleviates oxidative stress in random skin flaps. (a) Immunohistochemical staining of proteins SOD1 in random skin flaps (200 magnification; scan bar 25 μ m). (b) Quantification of integral absorbance of SOD1 in IHC. (c) Western blotting result of SOD1, eNOS, and HO1 in the control and CAT group. (d–f) Quantification of SOD1, eNOS, and HO1 expressions in each group. Significance: ∗∗ p < 0.01 vs. the control group. Data were expressed as means ± SEM, n = 6.

Article Snippet: In the molecular studies, the following antibodies were used: anti-Cadherin 5 (A02632-2; Boster Biological Technology, Wuhan, China), anti-GAPDH (AP0063; Biogot Technology, Shanghai, China), anti-VEGF (A12303, ABclonal, China), anti-SIRT1, anti-LC3II, anti-Superoxide Dismutase 1 (SOD1), anti-Cathepsin D (CTSD), anti-Caspase 3 (CAPS3), and anti-Heme Oxygenase 1 (HO1) (13161-1-AP, 14600-1-AP, 10269-1-AP, 21327-1-AP, 19677-1-AP, and 27282-1-AP; Proteintech Group, Chicago, IL, USA); the primary antibody anti-Bax, anti-Bcl-2, anti-AMPK, anti-p-AMPK, anti-mTOR, anti-p-mTOR, anti-Cleaved-Caspase 3 (C-CAPS3), and antiendothelial nitric oxide synthase (eNOS) were obtained from Cell Signaling Technology (CST) (2772S, 15071S, 5832S, 2535S, 2983, 5536S, 9664S, and 32027S; Beverly, MA, USA); the primary antibody anti-SQSTM1/p62, anti-CD34 and anti-Matrix Metalloproteinase 9 (MMP9) were obtained from Abcam(ab56416, ab81289, ab283575; Cambridge, UK).

Techniques: Immunohistochemical staining, Staining, Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Catalpol Enhances Random-Pattern Skin Flap Survival by Activating SIRT1-Mediated Enhancement of Autophagy

doi: 10.1155/2022/5668226

Figure Lengend Snippet: The beneficial effects of CAT on random flap viability are abolished by 3MA. (a, b) Representative images of flap necrotic area on days 3 and 7 in CAT and CAT+3MA groups after the surgery. Quantification of the percentage of survival area is shown in the histogram. (c, d) Images of the inner side of flaps in the CAT and CAT+3MA groups on day 7 after the surgery. Quantitative analysis of percentage of tissue water content in two groups. (e, f) LDBF images and quantitative analysis of flap blood flow in CAT and CAT+3MA groups. (g, h) H&E staining analysis and quantitative analysis of subcutaneous blood vessels of flaps in the CAT and CAT+3MA groups (200 magnification; scale bar, 50 μ m). (i, j) Immunohistochemical staining and quantitative analysis of CD34-positive vessels in CAT and CAT+3MA groups. (k, l) Immunofluorescence staining and quantitative analysis of LC3II-positive cells in dermal layer (scan bar, 10 μ m). (m, n) Western blotting and quantification analysis of autophagy proteins Belin1, LC3II, CTSD, and p62 in flaps after treating CAT with or without 3-MA. (o–r) Western blotting and quantification analysis of angiogenesis-related protein VEGFA and MMP9, oxidative stress-related protein SOD1 and HO1, and apoptosis-related protein Bax and Bcl-2 in each group. Significance: ∗ p < 0.05 and ∗∗ p < 0.01 vs. the CAT group. Data were expressed as means ± SEM, n = 6.

Article Snippet: In the molecular studies, the following antibodies were used: anti-Cadherin 5 (A02632-2; Boster Biological Technology, Wuhan, China), anti-GAPDH (AP0063; Biogot Technology, Shanghai, China), anti-VEGF (A12303, ABclonal, China), anti-SIRT1, anti-LC3II, anti-Superoxide Dismutase 1 (SOD1), anti-Cathepsin D (CTSD), anti-Caspase 3 (CAPS3), and anti-Heme Oxygenase 1 (HO1) (13161-1-AP, 14600-1-AP, 10269-1-AP, 21327-1-AP, 19677-1-AP, and 27282-1-AP; Proteintech Group, Chicago, IL, USA); the primary antibody anti-Bax, anti-Bcl-2, anti-AMPK, anti-p-AMPK, anti-mTOR, anti-p-mTOR, anti-Cleaved-Caspase 3 (C-CAPS3), and antiendothelial nitric oxide synthase (eNOS) were obtained from Cell Signaling Technology (CST) (2772S, 15071S, 5832S, 2535S, 2983, 5536S, 9664S, and 32027S; Beverly, MA, USA); the primary antibody anti-SQSTM1/p62, anti-CD34 and anti-Matrix Metalloproteinase 9 (MMP9) were obtained from Abcam(ab56416, ab81289, ab283575; Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Immunofluorescence, Western Blot