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: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Information for immunofluorescence antibodies and other reagents.

Article Snippet: Col 3 (α1) 488-Goat anti-rabbit DAPI , Servicebio Servicebio Servicebio , GB13023-2 GB25303 G1012 , 1:400 1:400.

Techniques: Immunofluorescence

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