Silencing MR-1 attenuates atherosclerosis in ApoE(−/−) mice induced by angiotensin II through FAK-Akt–mTOR-NF-kappaB signaling pathway

Myofibrillogenesis regulator-1 (MR-1) is a novel protein involved in cellular proliferation, migration, inflammatory reaction and signal transduction. However, little information is available on the relationship between MR-1 expression and the progression of atherosclerosis. Here we report atheroprotective effects of silencing MR-1 in a model of Ang II-accelerated atherosclerosis, characterized by suppression focal adhesion kinase (FAK) and nuclear factor kappaB (NF-κB) signaling pathway, and atherosclerotic lesion macrophage content. In this model, administration of the siRNA-MR-1 substantially attenuated Ang II-accelerated atherosclerosis with stabilization of atherosclerotic plaques and inhibited FAK, Akt, mammalian target of rapamycin (mTOR) and NF-kB activation, which was associated with suppression of inflammatory factor and atherogenic gene expression in the artery. In vitro studies demonstrated similar changes in Ang II-treated vascular smooth muscle cells (VSMCs) and macrophages: siRNA-MR-1 inhibited the expression levels of proinflammatory factor. These studies uncover crucial proinflammatory mechanisms of Ang II and highlight actions of silencing MR-1 to inhibit Ang II signaling, which is atheroprotective.

Expression of vascular basement membrane-derived multifunctional peptide in Pichia pastoris and identification of its bioactivity / 中国肿瘤生物治疗杂志

Objective: To express vascular basement membrane-derived multiple peptide (VBMDMP) in Pichia pastoris and to identify its bioactivity. Methods: PCR technique was used to obtain the target fragment GST-VBMDMP, which was then cloned into pPIC9K vector. The resultant vector pPIC9K-GST-VBMDMP was transfected into Pichia pastoris cells with spheroplast and the positive recons was identified. The His + Muts transformant was shake-cultured in MGY at 30 ℃. The culture supernatant (1 ml) was collected for preservation at-70 ℃ every 24 hours while maintaining the concentration of methanol at 0.5% in the culture medium. SDS-PAGE analysis was used to detect the protein expression after 8 days and then animal and cell experiments were performed with GST-VBMDMP. Results: SDS-PAGE analysis found that protein express increased during 1-7 days and decreased after 8 days in MGY medium; GST-VBMDMP fused protein was obtained by Glutathione Sepharose 4B and it inhibited the artery endothelial cell tube structure formation in C57BL/6 mouse; GST-VBMDMP( 2, 6, 10 mg/kg)also significantly inhibited the primary cancer Lewis mouse (tumor inhibitor rates being 96.6%, 82.1%, and 61.2%, respectively)and metastatic lung tumors(tumor inhibitor rates being 96.8 %, 87.9 %, and 75.3%, respectively)(P