An Experimental Study on the Establishment of Pulmonary Hypertension Model in Rats induced by Monocrotaline

Pulmonary hypertension is called PH for short. It is caused by the pulmonary artery vascular disease leading to pulmonary vascular resistance, and the increase right lung compartment load, which resulting in weakening or even collapse of the right ventricular function. The establishment of rat PH model under the action of monocrotaline is a repeatable, simple and accessible operation technique, which has been widely used in the treatment of pulmonary hypertension. This paper discusses the principle and properties of the PH model on rats under the monocrotaline action

Modulation of platelet-derived microparticles to adhesion and motility of human rheumatoid arthritis fibroblast-like synoviocytes.

Platelet-derived microparticles (PMPs) are closely associated with disease activity in rheumatoid arthritis (RA) and contribute to the inflammatory process. Rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) play important roles in the progression of joint destruction. The aim of this study is to demonstrate whether PMPs affect the adhesion and motility of RA-FLSs. Our data indicated that PMPs promoted migration, invasion and adhesion to extracellular matrix (ECM) of RA-FLSs. Further study showed that PMPs up-regulated the expression of matrix metalloproteinase-1 (MMP1) and increased the level of phosphorylation of NF-κB (p-NF-κB) and Erk (p-Erk) in RA-FLSs. These results suggest that PMPs promote RA-FLSs adhesion and motility presumably by increasing MMP1 via activating Erk-mediated NF-κB pathway

Effects of PMPs on adhesion of RA-FLSs.

<p>RA-FLSs were seeded onto the collagen I- (a), fibronectin- (b) or matrigel- (c) coated 96-well culture plates, respectively, and incubated at 37°C for 45 min. The CCK-8 assays were conducted to quantify the number of the adhesive cells by absorbance at 450 nm, and performed in six wells from four independent experiments. (*<i>p <</i> 0.05 vs. 0 μg/ml PMPs).</p

Effects of PMPs on the expression and activation of Erk and Akt in RA-FLSs.

<p>(A and B) Protein levels of p-Erk, Erk, Akt and p-Akt were analyzed from PMPs-treated RA-FLSs with corresponding antibodies. Quantification analysis was shown and expressed as fold-change. (*p < 0.05 vs. 0 μg/ml PMPs) (C) Quantification of the migration assay of RA-FLSs with PMPs and PD98059 was shown. (*<i>p <</i> 0.05 vs. PMPs) (D and E) Western blotting was performed to detect Erk, p-Erk, IκB, p- IκB, NF-κB and p- NF-κB of RA-FLSs after treated with PMPs and PD98059. (*<i>p</i> < 0.05 vs. PMPs) (F)RT-qPCR was conducted to detect the expression of MMP1 in RA-FLSs after treated with PMPs and PD98059. (*<i>p</i> < 0.05 vs. PMPs).</p

Verification of PMPs by flow cytometry.

<p>PMPs were isolated from platelet-rich plasma and stained with PE-labeled anti-CD41.The events in P2 were PMPs, which were compared to 0.82 μm standard microspheres.</p

Effects of PMPs on the expression and activation of IκB and NF-κB in RA-FLSs.

<p>(A and B) Western blotting was performed to detect the expression of p-IκB, IκB, NF-κB and p-NF-κB of RA-FLSs after treated with PMPs. The quantification was expressed as fold-change of 0μg/ml PMPs. (*<i>p</i> < 0.05 vs. 0 μg/ml PMPs) (C) Quantification of the migration assay of RA-FLSs treated with PMPs and JSH-23 was shown. (*<i>p <</i> 0.05 vs. PMPs) (D and E) After treated with PMPs and JSH-23, the expression of p-IκB, IκB, p-NF-κB and NF-κB of RA-FLSs were detected by western blot. (*<i>p</i> < 0.05 vs. PMPs) (F) After treated with PMPs and JSH-23, the expression of MMP1 was detected by RT-qPCR. (*<i>p</i> < 0.05 vs. PMPs).</p

Effects of PMPs on gene expression of RA-FLSs.

<p>(A) The clustering analysis compared the differences of gene expression between the group without (A6647) and with 50 μg/ml (A6648) PMPs: Red region, genes up-regulated in RA-FLSs. Green region, genes down-regulated in RA-FLSs. (B) Top 10 differential genes of pathway enrichment analysis are listed. (C) Relative mRNA levels of MMP1 and MMP2 of RA-FLSs after treated with PMPs were measured by RT-qPCR and GAPDH was used as an equal loading control. (*<i>p</i> < 0.05 vs. 0 μg/ml PMPs). (D) Protein levels of MMP1 and MMP2 were detected by western blot and normalized to GAPDH. (E) Quantification of protein levels was shown (*<i>p</i> < 0.05 vs. 0 μg/ml PMPs).</p