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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) <t>IL-6,</t> and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.
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Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) IL-6, and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.

Journal: Asian Journal of Andrology

Article Title: Near-infrared photobiomodulation therapy for age-related erectile dysfunction: molecular and physiological restoration in a mouse model

doi: 10.4103/aja202532

Figure Lengend Snippet: Histological analysis of corpus cavernosum tissue fibrosis and comprehensive analysis of erectile and senescence markers contents in different groups. ( a ) Penis samples in the young with NL, young with NIR, aged with NL, and aged with NIR were prepared for the detection of corpus cavernosum tissue fibrosis using Masson’s trichrome staining. Semi-quantitative image analysis of ( b ) smooth muscle/collagen ratio, ( c ) p21 expression, ( d ) γ-H2A.X expression, and ( e ) eNOS expression in corpus cavernosum tissues. ( f ) Representative western blotting images depicting the protein bands of interest. Quantitative assessment of relative protein expression levels for ( g ) eNOS, ( h ) α-SMA, ( i ) cell cycle inhibitor p16, ( j ) cell cycle inhibitor p21, ( k ) γ-H2A.X, ( l ) IL-6, and ( m ) IL-8. Data are expressed as the mean±standard error of the mean of 6 mice. Statistical analysis was performed using an unpaired t -test. * P <0.05, ** P <0.01, and *** P <0.001. NS: not significant; NL: natural light; NIR: near-infrared; γ-H2A.X: phosphorylated H2A histone family member X; eNOS: endothelial nitric oxide synthase; α-SMA: alpha-smooth muscle actin; IL-6: interleukin-6; IL-8: interleukin-8; AOD: average optical density.

Article Snippet: The polyvinylidene fluoride membrane was incubated with primary antibodies, including mouse anti-alpha-smooth muscle actin (anti-αSMA; 1:2000; ab7817; Abcam, Cambridge, UK), rabbit anti-eNOS (1:1000; #32027; CST, Danvers, MA, USA), rabbit anti-γ-H2A.X (1:4000; ab11175; Abcam,), rabbit anti-p21 (1:2000; 10355-1-AP; Proteintech, Wuhan, China), rabbit anti-p16 inhibitor of cyclin-dependent kinase 4 (INK4A) (1:3000; 10883-1-AP; Proteintech), rabbit anti-interleukin (IL)-6 (1:3000; 66146-1-Ig; Proteintech), and rabbit anti-IL-8 (1:1000; 27095-1-AP; Proteintech), at 4°C overnight.

Techniques: Staining, Expressing, Western Blot