Journal: Heliyon

Article Title: Quantitative proteomics reveals potential anti-inflammatory protein targets of radial extracorporeal shock wave therapy in TNF-α-induced model of acute inflammation in primary human tenocytes

doi: 10.1016/j.heliyon.2022.e12008

Figure Lengend Snippet: Immunofluorescence staining and rESWT effects on cell proliferation and pro-inflammatory cytokines. (A) Primary tenocytes immunostained for COL1 (A-1), COL3 (A-2), vimentin (A-3), SCX (A-4), OCT4 (A-5), and CD34 (A-6). (B) Changes in OD values (B-1) and IL-1β levels (B-2) at different time points after intervention with different concentrations of TNF-α. Data are expressed as means ± SD and were analyzed using one-way ANOVA ( n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus the control. # p < 0.05, ## p < 0.01, ### p < 0.001 indicate intragroup comparisons at different time points. (C) Effects of rESWT on tenocyte viability and reversal of TNF-α-induced decline in tenocyte proliferation and increase in IL-1β levels. (C-1) Effects of rESWT waves on tenocyte viability. (C-2) rESWT reversal of TNF-α-induced decline in tenocyte proliferation. (C-3) rESWT reversal of TNF-α-induced increase in IL-1β. Data are expressed as means ± SD and were analyzed using one-way ANOVA ( n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus the control. ▲ p < 0.05, ▲▲ p < 0.01, ▲▲▲ p < 0.001 indicate intergroup comparisons (except the control). # p < 0.05, ## p < 0.01, ### p < 0.001 indicate intragroup comparisons at different time points.

Article Snippet: Anti-COL1 (code: bsm-33400M, lot: BJ06239206), anti-COL3 (code: bs-0948R, lot: AH10092472), anti-vimentin (code: bs-0756R, lot: AI07165538), and anti-SCX (code: bs-12364R, lot: AC08264578) antibodies were all purchased from Bioss (Beijing, China).

Techniques: Immunofluorescence, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Inhibiting Lysyl Oxidases prevents pathologic cartilage calcification.

doi: 10.1016/j.biopha.2023.116075

Figure Lengend Snippet: Fig. 6. Proposed mechanism based on the observed results. Increased LOX(L) expression and activity promotes cartilage calcification (Anx, Enpp1, Pit1, Pit2, ALP). Both classical LOX(L) role (matrix cross-links) and non-classical roles are involved. The latter encompass: increased chondrocyte hypertrophy (Runx2, COL10), increased fibrosis (Col1, Col3), increased inflammation (IL-6) and increased oxidative stress (ROS). Finally, cartilage calcification can in turn induce increased LOX (L) activity.

Article Snippet: Collagen 1 (COL1) and Collagen 3 (COL3) expression were evaluated using an anti-COL1 rabbit polyclonal antibody (Boster, PA2140–2) and an anti-COL3 rabbit polyclonal antibody (Origene, AP06517PU-N) respectively.

Techniques: Expressing, Activity Assay

Journal: Acta Cirúrgica Brasileira

Article Title: Type I and type III collagen immunoexpression in rabbit skin biopsy samples treated with rosuvastatin gel and autologous platelet-rich plasma

doi: 10.1590/acb402725

Figure Lengend Snippet: Immunohistochemical analysis of the lesions. (a) Immunohistochemical staining for type I collagen on day 17 in the NC and the different treatments. Magnification: 400x. Scale bar: 50 μm. (b) Immunohistochemical staining for type III collagen on day 17 in the NC and the different treatments.

Article Snippet: Antibodies for evaluating collagen immunoexpression in rabbit wounds treated with aPRP, RSV, and aPRP + RSV included: anti-type I collagen primary antibody (1:50, mouse monoclonal, GeneTex, GTX26308), anti-type III collagen primary antibody (1:100, mouse monoclonal, Novus Biologicals, NBP1-05119), and goat anti-mouse IgG H&L (HRP) antibody (1:200, goat, Abcam, ab6789).

Techniques: Immunohistochemical staining, Staining

Journal: Acta Cirúrgica Brasileira

Article Title: Type I and type III collagen immunoexpression in rabbit skin biopsy samples treated with rosuvastatin gel and autologous platelet-rich plasma

doi: 10.1590/acb402725

Figure Lengend Snippet: Plots representing the average percentage of immunostaining for types I and III collagen under different treatments on day 17.

Article Snippet: Antibodies for evaluating collagen immunoexpression in rabbit wounds treated with aPRP, RSV, and aPRP + RSV included: anti-type I collagen primary antibody (1:50, mouse monoclonal, GeneTex, GTX26308), anti-type III collagen primary antibody (1:100, mouse monoclonal, Novus Biologicals, NBP1-05119), and goat anti-mouse IgG H&L (HRP) antibody (1:200, goat, Abcam, ab6789).

Techniques: Immunostaining

Journal: Frontiers in Immunology

Article Title: Induction of hepatic fibrosis in mice with schistosomiasis by extracellular microRNA-30 derived from Schistosoma japonicum eggs

doi: 10.3389/fimmu.2024.1425384

Figure Lengend Snippet: miRNA-30 derived from S. japonicum egg exosomes activates HSCs. (A) Exosomes were observed under transmission electron microscopy (TEM). (B) LX-2 cells were transfected with miRNA mimics for 48 h, followed by qPCR analysis to measure the mRNA levels of α-SMA . (C) LX-2 and mHSC cells were transfected with 50 nM of both NC mimic and miRNA-30 mimic, for 24 (h) Relative transcript levels of miRNA-30 were determined by qPCR. (D) LX-2 and mHSC cells were transfected with varying doses of NC mimic and miR-30 mimic for 48 h, followed by qPCR to quantify the mRNA levels of α-SMA , Col1(α1) , and Col3(α1) . (E) LX-2 cells were exposed to a 100 nM of both NC mimic and miRNA-30 mimic for 48 (h) The LX-2 cells were then subjected to Western blot analysis to detect their α-SMA protein expression levels. Both mRNA and protein levels of α-SMA , Col1(α1) , and Col3(α1) were normalized against GAPDH . GAPDH , 37 kDa; α-SMA , 43 kDa. NC, negative control; 30 mimic, miRNA-30 mimic. The results are averaged from three independent experiments. Student’s t -tests were carried out to assess differences between the two groups (* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: The membrane was then incubated with rabbit anti-GAPDH (5174S, CST, USA), anti-Col1(α1) (bs-10423R, Bioss Antibodies, USA), anti-Col3(α1) (bs-0548R, Bioss Antibodies), or anti-α-SMA (19245S, CST, USA) antibodies overnight at 4°C, followed by three 1×TBST washes.

Techniques: Derivative Assay, Transmission Assay, Electron Microscopy, Transfection, Western Blot, Expressing, Negative Control

Journal: Frontiers in Immunology

Article Title: Induction of hepatic fibrosis in mice with schistosomiasis by extracellular microRNA-30 derived from Schistosoma japonicum eggs

doi: 10.3389/fimmu.2024.1425384

Figure Lengend Snippet: Overexpression of miRNA-30 induces hepatic fibrosis in naive mice. Healthy six-week-old female C57BL/6 mice were injected with of PBS, rAAV8-SCR (blank vector), and rAAV8-miR-30 (overexpression vector) into the tail vein, with a viral titer of 5 × 10 or 1 × 10 vector genomes per mouse. (A) Liver and serum samples were collected three weeks post-injection to measure the miRNA-30 expression through qPCR, with normalization to U6 RNA ( n = 6). (B) Liver tissues harvested on days 49, 63, and 77 post-injection were analyzed for hydroxyproline content ( n = 6). (C–E) qPCR to quantify the mRNA levels of α-SMA , Col1(α1) , and Col3(α1) ( n = 6). (F–H) Western blot to quantify the protein levels of α-SMA , Col1(α1) , and Col3(α1) ( n = 6). Both mRNA and protein levels of α-SMA , Col1(α1) , and Col3(α1) were normalized against GAPDH . GAPDH , 37 kDa; α-SMA , 43 kDa; Col1(α1) , 130 kDa; Col3(α1) , 117 kDa. rAAV8, recombinant adeno-associated virus serotype 8; SCR, scrambled. Statistical significance between two or more groups was evaluated using Student’s t -test and one-way ANOVA (* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: The membrane was then incubated with rabbit anti-GAPDH (5174S, CST, USA), anti-Col1(α1) (bs-10423R, Bioss Antibodies, USA), anti-Col3(α1) (bs-0548R, Bioss Antibodies), or anti-α-SMA (19245S, CST, USA) antibodies overnight at 4°C, followed by three 1×TBST washes.

Techniques: Over Expression, Injection, Plasmid Preparation, Expressing, Western Blot, Recombinant, Virus

Journal: Frontiers in Immunology

Article Title: Induction of hepatic fibrosis in mice with schistosomiasis by extracellular microRNA-30 derived from Schistosoma japonicum eggs

doi: 10.3389/fimmu.2024.1425384

Figure Lengend Snippet: Inhibition of miRNA-30 from S. japonicum egg-derived exosomes delays the progression of hepatic fibrosis in schistosomiasis in mice. Mice received an injection of rAAV8-SCR or rAAV8-anti-miR-30-sponge vectors at a dose of 1 × 10 virus genomes or PBS via the tail vein. After 3 weeks, these mice were dermally exposed to 20 ± 1 S. japonicum cercariae. Liver samples were collected at days 42 and 56 post-infection. (A) Granulomas and collagen deposition were assessed through Masson’s trichrome staining. (B) Expression of α-SMA at the protein level was examined on day 42 post-infection through Western blot analysis. (C) Egg count in mouse liver tissue showing there was no statistical difference between the two groups ( n = 6). (D, E) Immunohistochemical analysis on days 42 and 56 post- S. japonicum infection revealed positively stained areas for Col1(α1) , Col3(α1) , and α-SMA . (F) qPCR to quantify the mRNA levels of α-SMA , Col1(α1) , and Col3(α1) ( n = 6). All images were magnified 10-fold. Quantification of α-SMA protein levels was normalized to GAPDH . GAPDH , 37 kDa; α-SMA , 43 kDa. rAAV8, recombinant adeno-associated virus serotype 8; SCR, scrambled. Statistical significance between two or more groups was evaluated using Student’s t -test and one-way ANOVA (* P < 0.05; ** P < 0.01; *** P < 0.001. ns, no significance.

Article Snippet: The membrane was then incubated with rabbit anti-GAPDH (5174S, CST, USA), anti-Col1(α1) (bs-10423R, Bioss Antibodies, USA), anti-Col3(α1) (bs-0548R, Bioss Antibodies), or anti-α-SMA (19245S, CST, USA) antibodies overnight at 4°C, followed by three 1×TBST washes.

Techniques: Inhibition, Derivative Assay, Injection, Virus, Infection, Staining, Expressing, Western Blot, Immunohistochemical staining, Recombinant

Journal: Europace

Article Title: Angiotensin II-mediated up-regulation of connective tissue growth factor promotes atrial tissue fibrosis in the canine atrial fibrillation model

doi: 10.1093/europace/eus052

Figure Lengend Snippet: Polymerase chain reaction primers used for amplification of inflammation and fibrosis-related genes

Article Snippet: After blocking with 5% non-fat milk, the membrane was incubated with anti-rabbit COL1 antibody (sc-8784; Santa Cruz Biotechnology, Inc., CA, USA), anti-rabbit COL3 antibody (LSL-LB-1387; Cosmo Bio Co., Ltd., Japan) or anti-mouse GAPDH antibody (GTX28245; GeneTex ® Inc., USA) and subsequently with the secondary antibody (Amersham ECL Plus Western blotting reagent pack; GE Healthcare UK Ltd.).

Techniques: Polymerase Chain Reaction, Amplification

Journal: Europace

Article Title: Angiotensin II-mediated up-regulation of connective tissue growth factor promotes atrial tissue fibrosis in the canine atrial fibrillation model

doi: 10.1093/europace/eus052

Figure Lengend Snippet: Messenger ribonucleic acid expressions of extracellular matrix and inflammation-related molecules. In the pacing control group, mRNA expressions of extracellular matrix-related genes (connective tissue growth factor, collagen type 1 and 3, and fibronectin 1) were up-regulated, especially in the right atrium in comparison with the non-pacing group, and this up-regulation was suppressed in the pacing + olmesartan group. In contrast, messenger ribonucleic acid expression of transforming growth factor-β did not differ among the three groups. See text for details. RA, right atrium; LA, left atrium; CTGF, connective tissue growth factor; COL1, collagen type 1; COL3, collagen type 3; FN1, fibronectin 1; TGF-β, transforming growth factor-β; MCP-1, monocyte chemotactic protein-1. * P < 0.05 vs. non-pacing group; † P < 0.05 vs. pacing control group.

Article Snippet: After blocking with 5% non-fat milk, the membrane was incubated with anti-rabbit COL1 antibody (sc-8784; Santa Cruz Biotechnology, Inc., CA, USA), anti-rabbit COL3 antibody (LSL-LB-1387; Cosmo Bio Co., Ltd., Japan) or anti-mouse GAPDH antibody (GTX28245; GeneTex ® Inc., USA) and subsequently with the secondary antibody (Amersham ECL Plus Western blotting reagent pack; GE Healthcare UK Ltd.).

Techniques: Control, Comparison, Expressing

Journal: Europace

Article Title: Angiotensin II-mediated up-regulation of connective tissue growth factor promotes atrial tissue fibrosis in the canine atrial fibrillation model

doi: 10.1093/europace/eus052

Figure Lengend Snippet: Western blot analysis of protein levels of collagens. ( A ) Representative examples of the bands and ( B ) a summary of all samples. Glyceraldehyde-3-phosphate dehydrogenase protein level was measured as an internal control. The protein level of collagen type 1 did not show any difference, but collagen type 3 was up-regulated in the pacing control group in comparison with the non-pacing group, and this up-regulation was suppressed in the pacing + olmesartan group. See text for details. RA, right atrium; LA, left atrium; COL1, collagen type 1; COL3, collagen type 3; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. * P < 0.05 vs. non-pacing group; † P < 0.05 vs. pacing control group.

Article Snippet: After blocking with 5% non-fat milk, the membrane was incubated with anti-rabbit COL1 antibody (sc-8784; Santa Cruz Biotechnology, Inc., CA, USA), anti-rabbit COL3 antibody (LSL-LB-1387; Cosmo Bio Co., Ltd., Japan) or anti-mouse GAPDH antibody (GTX28245; GeneTex ® Inc., USA) and subsequently with the secondary antibody (Amersham ECL Plus Western blotting reagent pack; GE Healthcare UK Ltd.).

Techniques: Western Blot, Control, Comparison

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Triptolide improves myocardial fibrosis in rats through inhibition of nuclear factor kappa B and NLR family pyrin domain containing 3 inflammasome pathway

doi: 10.4196/kjpp.2021.25.6.533

Figure Lengend Snippet: Primer sequence

Article Snippet: The prepared myocardial tissues of rats were sectioned, dewaxed, repaired in antigen repair solution, added normal goat serum sealing solution (C-0005; Shanghai Haoran Biotechnology Co., Ltd., Shanghai, China), and placed at room temperature for 20 min. Next, the sections were added primary antibodies rabbit anti-mouse NLRP3 (ab214185, 1:200; Abcam, Cambridge, MA, USA), ASC (ab47092, 1:100; Abcam), TGF-β1 (ab215715, 1:500; Abcam), COL1 (ab3453, 1:100; Abcam) and COL3 (ab23445, 1:100; Abcam) overnight at 4°C.

Techniques:

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Triptolide improves myocardial fibrosis in rats through inhibition of nuclear factor kappa B and NLR family pyrin domain containing 3 inflammasome pathway

doi: 10.4196/kjpp.2021.25.6.533

Figure Lengend Snippet: (A) H&E staining and Masson staining were used to observe the pathological morphology and MF of rats in each group (×200); (B) Masson staining results were quantitatively analyzed by measuring collagen volume fraction; (C) RT-qPCR was used to detect the expressions of TGF-β1, COL1, and COL3; (D) The expressions of TGF-β1, COL1, and COL3 were detected by immunohistochemistry; n = 6. Three independent repeated tests were performed and the data were expressed as mean ± SD; one-way was used for variance analysis; Tukey’s multiple comparisons test was used for post-hoc test. TPL, triptolide; MF, myocardial fibrosis; DMSO, dimethyl sulfoxide. Compared with sham group, **p < 0.01; compared with MF group, # p < 0.05, ## p < 0.01.

Article Snippet: The prepared myocardial tissues of rats were sectioned, dewaxed, repaired in antigen repair solution, added normal goat serum sealing solution (C-0005; Shanghai Haoran Biotechnology Co., Ltd., Shanghai, China), and placed at room temperature for 20 min. Next, the sections were added primary antibodies rabbit anti-mouse NLRP3 (ab214185, 1:200; Abcam, Cambridge, MA, USA), ASC (ab47092, 1:100; Abcam), TGF-β1 (ab215715, 1:500; Abcam), COL1 (ab3453, 1:100; Abcam) and COL3 (ab23445, 1:100; Abcam) overnight at 4°C.

Techniques: Staining, Quantitative RT-PCR, Immunohistochemistry

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Triptolide improves myocardial fibrosis in rats through inhibition of nuclear factor kappa B and NLR family pyrin domain containing 3 inflammasome pathway

doi: 10.4196/kjpp.2021.25.6.533

Figure Lengend Snippet: (A) RT-qPCR was used to detect the mRNA expressions of inflammatory factors IL-1β, IL-18, and TNF-α in myocardial tissues; (B) RT-qPCR was used to detect the mRNA expressions of TGF-β1, COL1, and COL3; (C) The expressions of TGF-β1, COL1, and COL3 were detected by immunohistochemistry (×200); n = 6. Three independent repeated tests were performed and the data were expressed as mean ± SD; independent t-test was used for comparisons between 2 groups. TPL, triptolide; MF, myocardial fibrosis; PBS, phosphate buffer saline. Compared with TPL + PBS group, **p < 0.01.

Article Snippet: The prepared myocardial tissues of rats were sectioned, dewaxed, repaired in antigen repair solution, added normal goat serum sealing solution (C-0005; Shanghai Haoran Biotechnology Co., Ltd., Shanghai, China), and placed at room temperature for 20 min. Next, the sections were added primary antibodies rabbit anti-mouse NLRP3 (ab214185, 1:200; Abcam, Cambridge, MA, USA), ASC (ab47092, 1:100; Abcam), TGF-β1 (ab215715, 1:500; Abcam), COL1 (ab3453, 1:100; Abcam) and COL3 (ab23445, 1:100; Abcam) overnight at 4°C.

Techniques: Quantitative RT-PCR, Immunohistochemistry

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Triptolide improves myocardial fibrosis in rats through inhibition of nuclear factor kappa B and NLR family pyrin domain containing 3 inflammasome pathway

doi: 10.4196/kjpp.2021.25.6.533

Figure Lengend Snippet: (A) WB was used to detect the expressions of p-P65 and t-P65 in myocardium nucleoprotein of rats in each group; (B) RT-qPCR was used to detect the mRNA expressions of inflammatory factors IL-1β, IL-18, and TNF-α in myocardial tissues of rats in each group; (C) RT-qPCR was used to detect the mRNA expressions of fibrosis related factors TGF-β1, COL1, and COL3 in myocardium; n = 6. Three independent repeated tests were performed and the data were expressed as mean ± SD; one-way was used for variance analysis; Tukey’s multiple comparisons test was used for post-hoc test. TPL, triptolide; MF, myocardial fibrosis; WB, Western blot; DMSO, dimethyl sulfoxide; PBS, phosphate buffer saline. Compared with TPL + PBS group, **p < 0.01. Compared with the MF group, ## p < 0.01.

Article Snippet: The prepared myocardial tissues of rats were sectioned, dewaxed, repaired in antigen repair solution, added normal goat serum sealing solution (C-0005; Shanghai Haoran Biotechnology Co., Ltd., Shanghai, China), and placed at room temperature for 20 min. Next, the sections were added primary antibodies rabbit anti-mouse NLRP3 (ab214185, 1:200; Abcam, Cambridge, MA, USA), ASC (ab47092, 1:100; Abcam), TGF-β1 (ab215715, 1:500; Abcam), COL1 (ab3453, 1:100; Abcam) and COL3 (ab23445, 1:100; Abcam) overnight at 4°C.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Huangqi Shengmai Yin Protects against Radiation-Induced Cardiac Fibrosis Injury by Regulating the TGF- β 1/Smads and MMPs

doi: 10.1155/2019/1358469

Figure Lengend Snippet: Regulation of HSY on TGF-β1/Smads signaling pathway and MMPs system in irradiated rats . A low dose (0.31 ml/d, LDG), medium dose (0.62 ml/d, MDG), and high dose (1.24 ml/d, HDG) of HSY were given daily to irradiated rats. (a) The protein expression of TGF- β 1, Col1, and Col3 in irradiated rats was analyzed with western blotting. (b) The protein expression of Smad 2, Smad 3, Smad 4, and Smad 7 in irradiated rats was analyzed with western blotting. (c) The protein expression of MMP14 and TIMP1 in irradiated rats was analyzed with western blotting. The average fluorescence intensity of each protein band is shown. All data are representative of three independent experiments. ## p<0.01 versus Con; ∗ p<0.05 versus 25 Gy; ∗∗ p<0.01 versus 25 Gy.

Article Snippet: Rabbit polyclonal anti-TGF- β 1, anti-Col1, anti-Col3, anti-Smad 2, anti-Smad3, anti-Smad4, and MMP14 were purchased from Gene Tex Corporation (Irvine, California, USA).

Techniques: Irradiation, Expressing, Western Blot, Fluorescence