microarray-based mirna platform  (Agilent technologies)

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Hispidin Inhibits Ferroptosis Induced by High Glucose via the miR-15b-5p/GLS2 Axis in Pancreatic Beta Cells

doi: 10.1155/2023/9428241

Figure Lengend Snippet: Hispidin treatment led to the upregulation of miR-15b-5p in PBCs. (a) PBCs treated with hispidin (40 μ M) or DMSO for 24 h; the expression of miRNAs was assayed using a microarray technique. (b) PBCs treated with hispidin (40 μ M) for 24 h; the expression of miR-15b-5p was analyzed by RT-PCR. PBCs treated with DMSO were used as the control. (c) PBCs transfected with the indicated miRNA mimic/inhibitor for 24 h; the expression of miR-802 was and analyzed by RT-PCR. (d)–(i) Under the HG condition, PBCs transfected with the indicated miRNA mimic/inhibitor for 24 h following which the cells were treated with hispidin (40 μ M) for an additional 24 h. Cells cultured under normal conditions were used as the control. (d) Cell viability measured. (e) Cell death measured. (f) ROS levels measured. (g) Fe 2+ levels measured. (h) MDA levels measured. (i) GSH levels measured. Data are representative of at least three independent experiments ( ∗ P < 0.05).

Article Snippet: The miRNA expression analysis was performed based on the Aksomics Technology (China) using the Agilent microarray-based miRNA platform.

Techniques: Expressing, Microarray, Reverse Transcription Polymerase Chain Reaction, Transfection, Cell Culture

Journal: Circulation

Article Title: Interleukin 10 Inhibits Bone Marrow Fibroblast Progenitor Cell-mediated Cardiac Fibrosis in Pressure Overloaded Myocardium

doi: 10.1161/CIRCULATIONAHA.117.027889

Figure Lengend Snippet: Total RNA was isolated from WT and IL10KO mice BM-FPCs using miRNeasy kit (Qiagen) and fibrosis-associated miRNA platform was used to assess the miRNAs level using Taqman microRNA assay. (A) At basal level fibrosis-associated miRNAs expression was significantly increased in IL10KO BM-FPCs compared to WT BM-FPCs with maximum expression of miRNA21, 145 and 208a (N=6). (B–D) miRNA21/145/208a expression was measured in WT-FPCs treated with IL10 and TGFβ by RT-PCR. IL10 treatment significantly inhibits TGFβ-induced miRNA-21, -145 and 208a expression. **p<0.01 vs BSA, #p<0.05 vs TGFβ. IL10KO mice: IL10 Knockout mice; BM-FPC: Bone marrow fibroblast progenitor cells. (N=6)

Article Snippet: Fibrosis-associated miRNA expression was analyzed in WT and IL10 KO BM-FPCs at basal level using a PCR-based miRNA microarray platform (Qiagen). miRNA-array data showed that several miRNAs were differentially expressed in IL10KO BM-FPCs compared to WT BM-FPCs ( ).

Techniques: Isolation, TaqMan microRNA Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Knock-Out

Journal: Circulation

Article Title: Interleukin 10 Inhibits Bone Marrow Fibroblast Progenitor Cell-mediated Cardiac Fibrosis in Pressure Overloaded Myocardium

doi: 10.1161/CIRCULATIONAHA.117.027889

Figure Lengend Snippet: BM-FPCs were transfected with miRNA-21 mimic for 12 hours using lipofectamine 2000 transfection reagent before IL10 and TGFβ treatments and miRNA-21 expression was measured by RT-PCR. (A) miRNA-21 mimic transfection significantly increased miRNA-21 expression in BM-FPCs. (B–E) 24 hrs after mimic, IL10 and TGFβ treatments, cells were harvested to measured the expression of fibrosis marker genes by RT-PCR. Relative levels of target genes expression were normalized with 18S. IL10 significantly inhibited TGFβ-induced fibrosis-associated genes expression. Restoration of miRNA-21 expression using miRNA-21 mimic significantly attenuated the IL10 effect on expression of these genes. ***p<0.001, **p<0.01, *p<0.05 vs BSA, ###p<0.001, ##p<0.01 vs TGFβ; $$$p<0.001, $p<0.05 vs TGFβ and ^^^p<0.001, ^^p<0.01, ^p<0.05 vs miR-21 mimic alone. (N=6) (F) Schematic diagram indicates TAC-induced mobilization and homing of BM-FPCs to the heart. In the heart, profibrotic stimuli (TGFβ in this case) first facilitates BM-FPC activation to myofibroblasts and then promote Smad2/3 dependent miRNA-21 maturation. The miRNA-21 maturation ultimately leads to fibrotic gene activation in activated myofibroblasts. On the other hand, IL10 inhibits TAC-induced mobilization, homing and activation of these cells and regulates cardiac fibrosis. Factor: Chemokine/cytokines; BMCs: Bone marrow cells; IL10 Interleukin-10.

Article Snippet: Fibrosis-associated miRNA expression was analyzed in WT and IL10 KO BM-FPCs at basal level using a PCR-based miRNA microarray platform (Qiagen). miRNA-array data showed that several miRNAs were differentially expressed in IL10KO BM-FPCs compared to WT BM-FPCs ( ).

Techniques: Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Marker, Activation Assay