Supernatants of Bifidobacterium longum and Lactobacillus plantarum Strains Exhibited Antioxidative Effects on A7R5 Cells

Vascular reactive oxygen species (ROS) play an essential role in cardiovascular diseases and the antioxidative effects of probiotics have been widely reported. To screen the probiotic strains that may prevent cardiovascular diseases, we tested the antioxidative effects of supernatants of different Bifidobacterium and Lactobacillus strains on A7R5 cells. Preincubation with supernatants of B. longum CCFM752, L. plantarum CCFM1149, or L. plantarum CCFM10 significantly suppressed the angiotensin II-induced increases in ROS levels and increased catalase (CAT) activity in A7R5, whereas CCFM752 inhibited NADPH oxidase activation and CCFM1149 enhanced the intracellular superoxide dismutase (SOD) activity simultaneously. Treatment with CCFM752, CCFM1149, or CCFM10 supernatants had no significant impact on transcriptional levels of Cat, Sod1, Sod2, Nox1, p22phox, or p47phox, but altered the overall transcriptomic profile and the expression of genes relevant to protein biosynthesis, and up-regulated the 60S ribosomal protein L7a (Rpl7a). A positive correlation between Rpl7a expression and intracellular CAT activity implied that Rpl7a may participate in CAT synthesis in A7R5. Supernatant of CCFM752 could also down-regulate the expression of NADPH oxidase activator 1 (Noxa1) and angiotensinogen in A7R5. Collectively, the probiotic strains CCFM752, CCFM1149, and CCFM10 exhibited antioxidative attributes on A7R5 cells and might help to reduce the risk of cardiovascular diseases.

Sabicea brasiliensis Wernham: Antioxidant Activity, Proliferative Effect and Modulation of Vascular Adenine Nucleotides Metabolism

Aims: Study addressed the antioxidant activity (AA) of Sabicea brasiliensis roots crude extract (CE), ethyl acetate (EAF), and hydro-methanolic (HMF) fractions, and its impact on cell viability and adenine nucleotide hydrolysis in vascular A7r5 cells. Materials and Methods: AA of CE, EAF and HMF were determined by the inhibition of the DPPH and ABTS radicals. Total phenolic content was evaluated by Folin-Ciocalteau. Cell viability was determined by MTT assay at different concentrations (62.5; 125; 250 and 500 μg·mL-1) of EAF and HMF after 24, 48 and 72 h. Ectonucleotidase activities were evaluated by colorimetric methods after 48 h EAF or HMF treatment. Results: The highest AA was observed for CE (76%), followed by EAF (46%) and HMF (23%). Phenolic content followed the same pattern. After 48 h, EAF increased A7r5 vascular cells viability by 40%, 40%, 62% and 25% at distinct concentrations, respectively; while HMF augmented it by 50% (500 μg·mL−1). Finally, after 48 h EAF (500 μg·mL−1) decreased about 50% of ATP and ADP metabolism while HMF inhibited 56 and 59% the hydrolysis of NPP substrate (at 125 and 250 μg·mL−1). Conclusion: Study confirmed the high AA of S. brasiliensis, which influences vascular cells proliferation and purines metabolism, pointing to potential cellular pathways that may support the popular use of this plant.

miR-129-5p inhibits oxidized low-density lipoprotein-induced A7r5 cells proliferation and migration through targeting HMGB1 involving a PI3K/Akt signal pathway

Background Currently, gene therapy for cardiovascular diseases has been widely concerned, but its mechanism is still unclear. Objective Recently miRNAs have been recognized as a key regulator in vascular smooth muscle cells (VSMCs) which involved in the formation of atherosclerosis. The aim of the study was to explore the role of miR-129-5p in regulation of HMGB1 involving a PI3K/Akt signal pathway as well as the proliferation and migration in A7r5 cells induced by ox-LDL. Methods Cell viability, proliferation and migration were conducted by CCK-8, colony formation, wound healing assay and transwell assay. The expressions of miR-129-5p and HMGB1 were detected by real-time quantitative-qPCR (RT-qPCR) and western blot. Luciferase assay was used to confirm that miR-129-5p directly targeted HMGB1. Results The expression of miR-129-5p in A7r5 cells induced by ox-LDL was significantly decreased in comparison with the control cells. Cell viability, proliferation and migration of A7r5 cells induced by ox-LDL were increased. MiR-129-5p could down-regulate the expression of HMGB1 in A7r5 cells. More studies showed that miR-129-5p could inhibit cell viability, proliferation on and migration of A7r5 cells induced by ox-LDL and target HMGB1 to regulate PI3K/Akt signal pathway. Conclusion miR-129-5p could inhibit PI3K/Akt signal pathway by target HMGB1 and further restrain the cell viability, proliferation and migration of A7r5 cells induced by ox-LDL.

FNDC5 Attenuates Oxidative Stress and NLRP3 Inflammasome Activation in Vascular Smooth Muscle Cells via Activating the AMPK-SIRT1 Signal Pathway

Vascular oxidative stress and inflammation play a major role in vascular diseases. This study was aimed at determining the protective roles of fibronectin type III domain-containing 5 (FNDC5) in angiotensin II- (Ang II-) induced vascular oxidative stress and inflammation and underlying mechanisms. Wild-type (WT) and FNDC5-/- mice, primary mouse vascular smooth muscle cells (VSMCs), and the rat aortic smooth muscle cell line (A7R5) were used in the present study. Subcutaneous infusion of Ang II caused more serious hypertension, vascular remodeling, oxidative stress, NLRP3 inflammasome activation, AMPK phosphorylation inhibition, and SIRT1 downregulation in the aorta of FNDC5-/- mice than those of WT mice. Exogenous FNDC5 attenuated Ang II-induced superoxide generation, NADPH oxidase 2 (NOX2) and NLRP3 upregulation, mature caspase-1, and interleukin-1β (IL-1β) production in A7R5 cells. The protective roles of FNDC5 were prevented by SIRT-1 inhibitor EX527, AMPK inhibitor compound C, or integrin receptor inhibitor GLPG0187. FNDC5 attenuated the Ang II-induced inhibition in SIRT1 activity, SIRT1 protein expression, and AMPKα phosphorylation in A7R5 cells, which were prevented by compound C, EX527, and GLPG0187. FNDC5 deficiency deteriorated Ang II-induced oxidative stress, NLRP3 inflammasome activation, AMPK phosphorylation inhibition, and SIRT1 downregulation in primary aortic VSMCs of mice, which were prevented by exogenous FNDC5. These results indicate that FNDC5 deficiency aggravates while exogenous FNDC5 alleviates the Ang II-induced vascular oxidative stress and NLRP3 inflammasome activation via the AMPK-SIRT1 signal pathway in VSMCs.

4-Pyridinio-1,4-Dihydropyridines as Calcium Ion Transport Modulators: Antagonist, Agonist, and Dual Action

A set of six new 4-pyridinio-1,4-dihydropyridine (1,4-DHP) compounds has been synthesized. The calcium channel modulating activity of these compounds was evaluated in an aorta vascular smooth muscle cell line (A7R5), in an isolated rat aortic ring model, and in human neuroblastoma cell lines (SH-SY5Y). The antagonistic effect of these 1,4-DHP was tested by modulating the impact of carbachol-dependent mobilization of intracellular Ca2+ in SH-SY5Y cells. The intracellular free Ca2+ concentration was measured in confluent monolayers of SH-SY5Y cells and A7R5 cells with the Ca2+-sensitive fluorescent indicator Fluo-4 NW. Only four compounds showed calcium channel blocking activity in SH-SY5Y and A7R5 cells as well as in the aortic ring model. Among them, compound 3 was the most active calcium channel antagonist, which had 3 times higher activity on carbachol-activated SH-SY5Y cells than amlodipine. Two of the compounds were inactive. Compound 4 had 9 times higher calcium agonist activity than the classic DHP calcium agonist Bay K8644. The intracellular mechanism for the action of compound 4 using inhibitor analysis was elucidated. Nicotinic as well as muscarinic receptors were not involved. Sarcoplasmic reticulum (ER) Ca2+ (SERCA) stores were not affected. Ryanodine receptors (RyRs), another class of intracellular Ca2+ releasing channels, participated in the agonist response evoked by compound 4. The electrooxidation data suggest that the studied compounds could serve as antioxidants in OS.

Seaweed Porphyra yezoensis polysaccharides with different molecular weights inhibit hydroxyapatite damage and osteoblast differentiation of A7R5 cells

Porphyra yezoensis polysaccharide could effectively reduce the cell damage and osteogenic transformation of vascular smooth muscle cells induced by hydroxyapatite crystals.