Journal: International Journal of Molecular Sciences

Article Title: The Implication of Substance P in the Development of Tendinopathy: A Case Control Study

doi: 10.3390/ijms18061241

Figure Lengend Snippet: Differential gene expressions between healthy tenocyte and tendinopathy tenocyte. Relative expressions by qRT-PCR of 8 genes were analyzed using cultured tenocytes. Results show that levels of COL3A1 , MMP1 , COX2 , SCX , ACTA2 , TAC1 ( SP ), and NK-1R were significantly increased in the cells cultured from tendinopathy tenocyte. However, COL1A1 was decreased in the cells cultured from tendinopathy tenocyte without statistical significance. The result was expressed as the relative expression level. Data were normalized on GAPDH mRNA levels. Boxplot showing the median along with the first and third quartiles. Error bar means data range from minimum to maximum. ** p < 0.01. SP: substance P; qRT-PCR: quantitative real time-polymerase chain reaction results.

Article Snippet: We used the following primary antibodies: COL3A1 protein 1:1000 (Gene Tex, Irvine, CA, USA), COX2 protein 1:1000 (Gene Tex), and MMP1 protein 1:1000 (Gene Tex).

Techniques: Quantitative RT-PCR, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: The Implication of Substance P in the Development of Tendinopathy: A Case Control Study

doi: 10.3390/ijms18061241

Figure Lengend Snippet: SP upregulation of genes related to tendinopathy. To determine the effects of SP on the development of tendinopathy, cultured healthy tenocytes were pretreated with or without SP 10 −7 M for 24 h. Results show that healthy tenocytes developed similar gene expression trends as that of the tendinopathy tenocyte. Levels of COL1A1 decreased but COL3A1 , COX2 , SCX , MMP1 , and ACTA2 were significantly increased as a result of treatment with SP. The result was expressed as the relative expression level after normalizing on GAPDH mRNA levels. Boxplot showing the median along with the first and third quartiles. Error bar means data range from minimum to maximum. * p < 0.05; ** p < 0.01; SP: substance P.

Article Snippet: We used the following primary antibodies: COL3A1 protein 1:1000 (Gene Tex, Irvine, CA, USA), COX2 protein 1:1000 (Gene Tex), and MMP1 protein 1:1000 (Gene Tex).

Techniques: Cell Culture, Gene Expression, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Implication of Substance P in the Development of Tendinopathy: A Case Control Study

doi: 10.3390/ijms18061241

Figure Lengend Snippet: SP induced the development of tendinopathy at protein level. Western blot for the cultured healthy tenocyte with or without SP was performed with antibodies against COL3A1 , COX2 , and MMP1 protein. Loading control was performed with an anti-β-actin antibody. Twenty-four hours of exposure to SP resulted in strong band densities for every protein. The results were consistent with the analysis of qRT-PCR ( A ); Densitometric analysis, as expected, revealed that COL3A1 , COX2 , and MMP1 protein band intensities normalized to β-actin were increased by 203.8%, 129.3%, and 129.3% respectively in SP-treated group compared with PBS-control. Boxplot showing the median along with the first and third quartiles ( B ). Error bar means data range from minimum to maximum. * p < 0.05; SP: substance P.

Article Snippet: We used the following primary antibodies: COL3A1 protein 1:1000 (Gene Tex, Irvine, CA, USA), COX2 protein 1:1000 (Gene Tex), and MMP1 protein 1:1000 (Gene Tex).

Techniques: Western Blot, Cell Culture, Control, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Therapeutic Effects of Long-Term Administration of Tranilast in an Animal Model for the Treatment of Fibroids

doi: 10.3390/ijms241310465

Figure Lengend Snippet: ( A ) Relative expression of COL3A1, CCND1, FN1, E2F1, and TGF-β3 mRNA in xenografts implanted subcutaneously in the ovariectomized CB-17 SCID/Beige mice (n = 16) following 8 weeks of treatment with the vehicle or tranilast (50 mg/kg/daily). ( B ) Representative Western blot analysis of COL3A1, CCND1, FN1, and E2F1 with bar graphs ( C ) showing their relative band densities in the xenografts (n = 16). ( D ) TGF-β3 levels as determined by enzyme-linked immunosorbent assay in 16 xenografts. The results are presented as mean ± SEM, with p values indicated by corresponding lines. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After transferring the samples to a nitrocellulose membrane, the membrane was blocked with TBS-Tween + 5% milk and probed with the following primary antibodies: COL3A1 and FN1 (Proteintech Group, Inc., Chicago, IL, USA), CCND1 (Cell Signaling Technology, Danvers, MA, USA), and E2F1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Therapeutic Effects of Long-Term Administration of Tranilast in an Animal Model for the Treatment of Fibroids

doi: 10.3390/ijms241310465

Figure Lengend Snippet: Schematic diagram representing the potential inhibitory mechanism of tranilast on fibroid growth and progression. Fibroids are characterized by increased expression levels of COL3A1, FN1, and TGF-β3 (ECM deposition and tissue fibrosis) [ , ] and CCND1 and E2F1 (mediated cell proliferation) [ , ]. Tranilast treatment significantly inhibits the expression of these genes while inducing apoptosis through increased levels of cleaved caspase 3, resulting in repressed ECM deposition, proliferation, and decreased fibroid size.

Article Snippet: After transferring the samples to a nitrocellulose membrane, the membrane was blocked with TBS-Tween + 5% milk and probed with the following primary antibodies: COL3A1 and FN1 (Proteintech Group, Inc., Chicago, IL, USA), CCND1 (Cell Signaling Technology, Danvers, MA, USA), and E2F1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing