Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: Expression of miR-145, ADAM17, ACE2, and Mas receptor in the aortic arteries of hypertensive rats. 22 male Wistar rats were selected randomly to receive standard diet or high-sucrose/high-fat diet for 30 weeks, the thoracic aorta was collected, the expression of ACE2 was detected by Western blotting, and the expression of MASR, miR-145, and ADAM17 was detected by qPCR. (a) Quantification of ACE2 expression from panel (b) data was expressed after normalization to the β -actin. (b) Representative figures from Western blotting of ACE2. (c–e) Quantification of mRNA expression of MASR, miR-145, and ADAM17 (qPCR). ∗∗ P < 0.01 between groups and ∗ P < 0.05 between groups.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: Expressing, Western Blot

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: miR-145 augments Ang II-induced ACE2-Ang-(1–7)-Mas axis activation and ADAM17 expression in VSMCs. VSMCs were treated with control, Ang II (1 μ M), miR-145 mimic (100 nM), or miR-145 (100 nM) inhibitor alone or in combination for 48 hours. (a) Concentration of Ang-(1–7) in the supernatant (ELISA). (b, c) ACE2 and MASR expression from panel (e) and data are expressed as the fold of the GAPDH. (d) ADAM17 expression in groups from panel (f). (e, f) Representative figures of ACE2, MASR, and ADAM17 expression in VSMCs (Western blotting). ∗∗ P < 0.01 vs. control group; ∗ P < 0.05 vs. control group; ## P < 0.01 vs. Ang II group; and # P < 0.05 vs. Ang II group. All the data are expressed as mean ± SEM from three independent experiments. miRNA NC, negative control miRNA.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: Activation Assay, Expressing, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Negative Control

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: ADAM17 siRNA reversed phenotype transition induced by miR-145 in vitro. VSMCs were treated with miR-145 inhibitor in the presence or absence of ADAM17 siRNA (100 nM) as indicated for 48 hours. The expression of α -SMA (a), SM22 α (b), OPN (c), EREG (d), and MMP2 (e) protein was detected by Western blotting. All the data were normalized to that of GAPDH. (f) Representative figures of OPN, α -SMA, SM22 α , EREG, and MMP2 (Western blotting). (g) The luciferase reporter assay is shown. Cells were transfected with a reporter vector psiCHECK-2-ADAM17 3′-UTR plus either miR-145-5p or the negative control. ∗∗ P < 0.01 vs. control group or miR-145-5p NC and ## P < 0.01 vs. miR-145 inhibitor group. All the data are expressed as mean ± SEM of three independent experiments. NC siRNA, negative control siRNA. WT, wide type. MT, mutant type.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: In Vitro, Expressing, Western Blot, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Negative Control, Control, Mutagenesis

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: ADAM17 mediated miR-145-induced effect by regulating ACE2-Ang-(1–7)-Mas axis in vitro. VSMCs were treated with miR-145 inhibitor in the presence or absence of ADAM17 siRNA (100 nM) as indicated for 48 hours. (a) Concentration of Ang-(1–7) in the supernatant (ELISA). (b) Representative figures of ACE2 and MASR (Western blotting). Quantification of ACE2 (c) and MASR (d) expression determined by Western blotting, and the data were normalized to that of GAPDH. ∗∗ P < 0.01 vs. control group; ∗ P < 0.05 vs. control group; and ## P < 0.01 vs. miR-145 inhibitor group. All the data are expressed as mean ± SEM of three independent experiments. NC siRNA, negative control siRNA.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control, Negative Control