Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: The expression levels of decorin were found reduced in muscles during ageing. (a) The forelimb grip strength and running times in young and aged group mice ( n = 6). (b) The ratio of gastrocnemius (Gast), quadriceps femoris (QU) and tibialis anterior (TA) weights/body weight ( n = 6). (c) Haematoxylin and eosin histological analysis and average cross‐sectional area of fibre sizes ( n = 6). (d) Dcn expression levels in young and aged mice in GSE132040 ( n = 3). (e) qRT‐PCR and Western blotting analysed the decorin protein and mRNA levels in gastrocnemius from 3‐ and 18‐month‐old mice ( n = 6). (f) Western blotting analysed the expression of p16INK4a, MuRF‐1, Atrogin‐1, α‐SMA and COL‐1 proteins ( n = 6). GAPDH: internal reference. Scale bar, 100 μm. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the young group. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Expressing, Muscles, Quantitative RT-PCR, Western Blot

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: Decorin deficiency in D‐gal–induced aged mice exacerbated skeletal muscle wasting. (a) The ratio of Gast/body weight in Dcn +/+ , Dcn +/+ –D‐gal and Dcn −/− –D‐gal groups ( n = 6). (b) The grip strength, muscular strength and running times of mice were analysed by electronic dynamometer, four‐limb hanging test and treadmill ( n = 6). (c) Representative H&E, fast muscle fibres and slow muscle fibres staining, quantifications of muscle fibre sizes distribution, average cross‐sectional areas (CSAs) and CSA of slow/fast muscle fibres ( n = 6). (d) Western blotting analysed the protein levels of α‐SMA, COL1, MuRF‐1, Atrogin‐1, MSTN, p16INK4a and p21 in Gast muscles ( n = 3). (e) Relative mRNA levels of IL‐6, IL‐1β and TNF‐α genes in Gast muscles. (f) Western blotting analysed the protein levels of NLRP3, p‐p65 and p65 in Gast muscles. Scale bar, 100 μm. Data are presented as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 compared with Dcn +/+ group. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Staining, Western Blot, Muscles

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: Dcn overexpression ameliorated age‐associated phenotypes of NOR‐10 cells induced by D‐gal. (a) SA‐β‐gal staining in NC–D‐gal and Dcn OE –D‐gal groups ( n = 3). (b) Protein levels of p53, p16INK4a, p21 and decorin in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups by Western blotting ( n = 3). (c) Protein levels of Atrogin‐1, MuRF‐1, MSTN, fibronectin and α‐SMA in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups by Western blotting; the relative protein levels of the target proteins were normalised to those of β‐actin ( n = 3). (d) Immunofluorescence staining of α‐SMA in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups ( n = 3). (e) Relative mRNA levels of IL‐6, IL‐1β and TNF‐α genes in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups. (f) Western blotting analysed the protein levels of NLRP3 and p‐p65 in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups. Scale bar, 100 μm. Data are presented as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 compared with NC and Dcn OE group. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Over Expression, Staining, Western Blot, Immunofluorescence

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: Recombinant decorin attenuated senescence and fibrosis in D‐gal–induced NOR‐10 cells and primary skeletal muscle cells. (a) Western blotting analysed the protein levels of p16INK4a, p53, p21 and decorin in Ctrl, D‐gal and D‐gal + decorin groups ( n = 3). (b) Western blotting analysed the protein levels of Atrogin‐1, MuRF‐1, MSTN, fibronectin and α‐SMA in Ctrl, D‐gal and D‐gal + decorin groups ( n = 3). (c) Relative mRNA levels of IL‐6, IL‐1β and TNF‐α genes in Ctrl, D‐gal and D‐gal + decorin groups. (d) Western blotting analysed the protein levels of NLRP3 and p‐p65 in Ctrl, D‐gal and D‐gal + decorin groups. (e) Western blotting analysed the protein levels of p53, p16INK4a and p21 in Ctrl, D‐gal and D‐gal + decorin groups. (f) Western blotting analysed the protein levels of PAX7 in Ctrl, D‐gal and D‐gal + decorin groups. Scale bar, 100 μm. Data are presented as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 compared with Ctrl group. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Recombinant, Western Blot

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: Akt/mTOR signalling pathway was involved in the regulation of decorin on skeletal muscle fibrosis and atrophy. (a) Western blotting analysed the expression levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K, p‐4E‐BP1, GAPDH and the corresponding statistical analysis in the Dcn +/+ , Dcn +/+ –D‐gal or Dcn −/− –D‐gal groups ( n = 3). (b) Western blotting analysed the expression protein levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K, p‐4E‐BP1 and β‐actin and the corresponding statistical analysis in the si‐NC, si‐Dcn, si‐NC–D‐gal and si‐Dcn–D‐gal groups ( n = 3). (c) Western blotting analysed the expression levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K, p‐4E‐BP1 and β‐actin and the corresponding statistical analysis in the NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups ( n = 3). (d) Western blotting analysed the expression levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K, p‐4E‐BP1 and β‐actin and the corresponding statistical analysis in the Ctrl, D‐gal and D‐gal + decorin groups ( n = 3). (e) Western blotting analysed the expression levels of p62, LC3b, GAPDH and the corresponding statistical analysis in the Dcn +/+ , Dcn +/+ –D‐gal or Dcn −/− –D‐gal groups ( n = 3). Data are presented as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 compared with Dcn +/+ , si‐NC, NC and Ctrl groups, respectively. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Western Blot, Expressing

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: ITGB1 was involved in the regulation of fibrosis and atrophy by decorin. (a) Western blotting analysed the expression levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K and β‐actin and the corresponding statistical analysis in Dcn + si‐NC, Dcn + si‐ITGB1, Dcn + si‐NC–D‐gal and Dcn + si‐ITGB1–D‐gal groups ( n = 3). (b) Western blotting analysed the effect of ITGB1 expression in NC, Dcn OE , NC–D‐gal and Dcn OE –D‐gal groups ( n = 3). (c) Western blotting analysed the effect of ITGB1 expression in NC, ITGB1 OE , NC–D‐gal and ITGB1 OE –D‐gal groups ( n = 3). (d) Lysates of NOR‐10 cells were immunoprecipitated with decorin antibody, and then Western blotting was performed with anti‐ITGB1 ( n = 3). (e) Representative images of decorin and ITGB1 expression in NOR‐10 cells obtained by co‐immunofluorescence staining ( n = 3). (f) Western blotting analysed the expression levels of Akt, p‐S473‐Akt, mTOR, p‐Ser2448‐mTOR, p70, p‐p70S6K and β‐actin and the corresponding statistical analysis in the NC, ITGB1 OE , NC–D‐gal and ITGB1 OE –D‐gal groups ( n = 3). Scale bar, 50 μm. Data are presented as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 compared with si‐NC or NC group. ns = no significant difference.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: Western Blot, Expressing, Immunoprecipitation, Immunofluorescence, Staining

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Decorin Deficiency Promotes D‐Galactose–Induced Skeletal Muscle Atrophy and Fibrosis by Regulating ITGB1/Akt/mTOR Signalling Pathway

doi: 10.1002/jcsm.70144

Figure Lengend Snippet: Schematic presentation of the decorin/ITGB1/Akt pathway.

Article Snippet: To test the effect of recombinant mouse decorin on D‐gal–induced senescent cells, different doses of decorin (Novoprotein, CM87) and the cells were co‐incubated with D‐gal for 72 h.

Techniques: