tsp1  (Athens Research)

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: LSKL suppresses TSP1/TGF-β/SMAD pathway and CC fibrosis in DM rats. (A) Representative immunoblot of TSP1, and active TGF-β1 in CC of the two groups. Semi-quantification of TSP1 (B) and active TGF-β1 (C) in CC of the two groups. (D) Representative immunoblot of p-SMAD2/3, SMAD2/3, and collagen IV in CC of the three groups after 4 weeks of treatment. Semi-quantification of the ratio of SMAD2/3 to their phosphorylated forms (E) and collagen IV (F) in CC of the three groups. Representative immunofluorescence (×200) (G) and semi-quantification (H) of collagen IV in CC of the three groups. Representative immunofluorescence (×200) (I) and semi-quantification (J) of fibronectin in CC of the three groups. a p<0.05, compared to control, b p<0.05, compared to DMED. DMED: diabetes mellitus-induced erectile dysfunction, TSP1: thrombospondin-1, TGF: transforming growth factor, LSKL: Leu-Ser-Lys-Leu, CC: corpus cavernosum, DM: diabetes mellitus.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Western Blot, Immunofluorescence, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: HG activates TSP1/TGF-β/SMAD pathway and induces fibrosis in CCSMCs. (A) Representative immunofluorescence of α-SMA in CCSMCs in intro. Representative immunoblot (B) and semi-quantification of TSP1 (C), and active TGF-β1 (D) in conditioned medium of CCSMCs. Representative immunoblot (E) and semi-quantification of TSP1 (F), The ratio of SMAD2/3 to their phosphorylated forms (G), SMAD2/3 (H), and collagen IV (I) in CCSMCs. a p<0.05, compared to control. α-SMA: α-smooth muscle actin, HG: high glucose, TGF: transforming growth factor, TSP1: thrombospondin-1, CC: corpus cavernosum, SMCs: smooth muscle cells.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Immunofluorescence, Western Blot, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: LSKL suppresses TSP1/TGF-β/SMAD pathway and fibrosis induced by TSP1 in CCSMCs. (A) Representative immunoblot of p-SMAD2/3, SMAD2/3, and collagen IV in CCSMCs of the four groups after TSP1 treatment. Semi-quantification of the ratio of SMAD2/3 to their phosphorylated forms (B), SMAD2/3 (C), and collagen IV (D) in CC of the four groups. (F) Representative immunoblot of p-SMAD2/3, SMAD2/3, and collagen IV in CCSMCs of the three groups after TSP1 and LSKL treatment. Semi-quantification of the ratio of SMAD2/3 to their phosphorylated forms (E), SMAD2/3 (G), and collagen IV (H) in CCSMCs of the three groups. Representative immunofluorescence (I) and semi-quantification (J) of collagen IV among the groups in CCSMCs. a p<0.05, compared to control; b p<0.05, compared to HG. TSP1: thrombospondin-1, LSKL: Leu-Ser-Lys-Leu, TGF: transforming growth factor, CC: corpus cavernosum, SMCs: smooth muscle cells.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Western Blot, Immunofluorescence, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: Blockade of TSP1 attenuates TGF-β/SMAD pathway and fibrosis induced by HG in CCSMCs. Representative immunoblot (A) and semi-quantification of TSP1 (B), the ratio of SMAD2/3 to their phosphorylated forms (C), and collagen IV (D) in CCSMCs. Representative immunofluorescence (×200) (E) and semi-quantification (F) of collagen IV among the groups in CCSMCs. Representative immunoblot (G) and semi-quantification of TSP1 (H), and active TGF-β1 (I) in conditioned medium of CCSMCs. Representative immunoblot (J) and semi-quantification of TSP1 (K), the ratio of SMAD2/3 to their phosphorylated forms (L), and collagen IV (O) in CCSMCs. Representative immunofluorescence (×200) (M) and semi-quantification (N) of collagen IV among the groups in CCSMCs. a p<0.05, compared to control; b p or c p<0.05, compared to HG. HG: high glucose, LSKL: Leu-Ser-Lys-Leu, TSP1: thrombospondin-1, TGF: transforming growth factor, CC: corpus cavernosum, SMCs: smooth muscle cells.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Western Blot, Immunofluorescence, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: Secretion of TSP1 induced by HG in CCFs stimulates fibrosis in CCSMCs. (A) Representative immunofluorescence of DCN in CCFs in intro. Representative immunoblot (B) and semi-quantification of TSP1 (C) in CCFs. Representative immunoblot (E) and semi-quantification of TSP1 (D), and active TGF-β1 (F) in conditioned medium of fibroblasts. Representative immunoblot (H) and the ratio of SMAD2/3 to their phosphorylated forms (I), SMAD2/3 (G), and collagen IV (J) in CCSMCs cultured by conditioned medium of CCFs. a p<0.05, compared to control. DCN: decorin, HG: high glucose, TSP1: thrombospondin-1, TGF: transforming growth factor, CCFs: CC fibroblasts, CC: corpus cavernosum, SMCs: smooth muscle cells.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Immunofluorescence, Western Blot, Cell Culture, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: Blockade of TSP1 in CCFs induced by HG attenuates fibrosis in CCSMCs. Representative immunoblot (A) and semi-quantification of TSP1 (B) in CCFs. Representative immunoblot (C) and semi-quantification of TSP1 (D), and active TGF-β1 (E) in conditioned medium of CCFs. Representative immunoblot (F) and the ratio of SMAD2/3 to their phosphorylated forms (G), SMAD2/3 (H), and collagen IV (I) in CCSMCs cultured by conditioned medium of CCFs. Data are expressed as means±standard deviation. a p<0.05, compared to control; b p or c p<0.05, compared to HG. HG: high glucose, TSP1: thrombospondin-1, TGF: transforming growth factor, CC: corpus cavernosum, SMCs: smooth muscle cells, CCFs: CC fibroblasts.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: Western Blot, Cell Culture, Standard Deviation, Control

Journal: The World Journal of Men's Health

Article Title: Blocking TSP1 Ameliorates Diabetes Mellitus-Induced Erectile Dysfunction by Inhibiting the TGF-β/SMAD Pathway

doi: 10.5534/wjmh.240065

Figure Lengend Snippet: Proposed mechanism of effects of TSP1 on DMED. STZ: streptozotocin, DMED: diabetes mellitus-induced erectile dysfunction, TGF-β1: transforming growth factor-β1, LSKL: Leu-Ser-Lys-Leu, TSP1: thrombospondin-1, CCFs: corpus cavernosum fibroblasts, CCSMCs: corpus cavernosum smooth muscle cells.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against TSP1 (1:1,000, #37879; Cell Signaling Technology), TGF-β1 (1:1,000, # AB179695 ; Abcam), phospho-SMAD2/3 (Ser1177) (1:500, #8685; Cell Signaling Technology), collagen IV (1:1,000, #19674-1-AP; Proteintech), SMAD2/3 (1:2,000, #8828; Cell Signaling Technology), and β-actin (1:10,000, #66009-1-Ig; Proteintech).

Techniques: