Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: GE‐activated ACE2 catalysed the conversion of Ang‐II to Ang‐(1–7) in hypoxic HCECs. HCECs were divided into normal, hypoxia, hypoxia + GE (40 μM for 24 h), and hypoxia + GE + MLN‐4760 (100 nM for 24 h) groups. (A) The protein expression of Ang II in HCECs was detected via ELISA. (B) The protein expression of Ang‐(1–7) in HCECs was detected via ELISA (** p < 0.01 vs. the normal group; # , p < 0.05 vs. the hypoxia group; % , p < 0.05 vs. the hypoxia + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: Ang‐(1–7) catalysed by GE‐activated ACE2 activated downstream MasR. HCECs were divided into normal, hypoxia, hypoxia + GE (40 μM for 24 h), hypoxia + GE + MLN‐4760, and hypoxia + GE + D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) groups. (A) The protein expression of MasR in HCECs was detected via western blotting. (B) Relative protein expression of MasR (** p < 0.01 vs. the normal group; # p < 0.05 vs. the hypoxia group; % p < 0.05 vs. the hypoxia + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

Techniques: Expressing, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: Activation of the ACE2/Ang‐(1–7)/MasR pathway by GE promoted the production and release of IL‐10 from hypoxic HCECs. HCECs were divided into normal, hypoxia, hypoxia + GE, hypoxia + GE + MLN‐4760, hypoxia + GE + D‐Pro7‐Ang‐(1–7), and hypoxia + GE + A‐779 (1 μM for 24 h) groups. (A) The protein expression of IL‐10 in HCECs was detected via Western blotting. (B) Relative protein expression of IL‐10. C IL‐10 levels in the culture supernatants of HCECs were assessed via ELISA (** p < 0.01 vs. the normal group; # p < 0.05 vs. the hypoxia group; % p < 0.05 vs. the hypoxia + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

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

Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: Activation of the ACE2/Ang‐(1–7)/MasR/IL‐10 pathway by GE suppressed the hypoxia‐induced proliferative, migratory, and tube‐forming abilities of HCECs. HCECs were divided into normal, hypoxia, hypoxia + GE, hypoxia + GE + MLN‐4760, hypoxia + GE + D‐Pro7‐Ang‐(1–7), hypoxia + GE + A‐779, and hypoxia + GE + neutralising anti‐IL‐10 antibody (2 μg for 24 h) groups. (A) EdU incorporation assay was performed to assess the proliferative ability of HCECs. (B) Ratio of the number of EdU‐positive cells to the number of DAPI‐positive cells. (C) Transwell assay was performed to assess the migratory ability of HCECs. (D) Number of migrated HCECs. (E) Tube formation assay was performed to assess the tube‐forming ability of HCECs. (F) Length of the tubes formed (all scale bars = 50 μm) (** p < 0.01 vs. the normal group; # p < 0.05; ## p < 0.01 vs. the hypoxia group; % p < 0.05 vs. the hypoxia + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

Techniques: Activation Assay, Transwell Assay, Tube Formation Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: GE activated the ACE2/Ang‐(1–7)/MasR/IL‐10 pathway in mice with laser‐induced CNV. Mice were divided into normal, CNV 7 d, CNV 7 d + GE (20 μM), CNV 7 d + GE + MLN‐4760 (10 nM), CNV 7 d + GE + D‐Pro7‐Ang‐(1–7) (100 nM), CNV 7 d + GE + A‐779 (100 nM), and CNV 7 d + GE + neutralising anti‐IL‐10 antibody (1 μg) groups. (A) The protein expression of ACE2 in the plasma membrane of mouse retina–RPE–choroid tissues was detected via western blotting. (B) Relative protein expression of ACE2. (C) The activity of ACE2 in the plasma membrane of mouse retina–RPE–choroid tissues was measured using an ACE2 activity detection kit. (D) The protein expression of Ang‐(1–7) in mouse retina–RPE–choroid tissues was detected via ELISA. (E) The protein expression of MasR in mouse retina–RPE–choroid tissues was detected via western blotting. (F) Relative protein expression of MasR. (G) The protein expression of IL‐10 in mouse retina–RPE–choroid tissues was detected via ELISA (** p < 0.01 vs. the normal group; # p < 0.05 vs. the CNV 7 d group; % p < 0.05 vs. the CNV 7 d + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

Techniques: Expressing, Clinical Proteomics, Membrane, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Geraniin Alleviates Mouse Laser‐Induced Choroidal Neovascularisation by Inhibiting Choroidal Endothelial Cell ACE2 /Ang‐(1–7)/ MasR / IL ‐10 Pathway

doi: 10.1111/jcmm.70228

Figure Lengend Snippet: GE suppressed the development of CNV by activating the ACE2/Ang‐(1–7)/MasR/IL‐10 pathway in mice. Mice were divided into normal, CNV 7 d, CNV 7 d + GE (20 μM), CNV 7 d + GE + MLN‐4760 (10 nM), CNV 7 d + GE + D‐Pro7‐Ang‐(1–7) (100 nM), CNV 7 d + GE + A‐779 (100 nM), and CNV 7 d + GE + neutralising anti‐IL‐10 antibody (1 μg) groups. (A) Representative images of choroidal flat mounts after triple staining with anti‐IB4 antibody (a vascular endothelium marker; green), anti‐collagen IV antibody (a basal lamina collagen marker; red), and DAPI (a nuclear dye; blue) (scale bar = 100 μm). (B) CNV volume ( ## p < 0.01 vs. the CNV 7 d group; % p < 0.05 vs. the CNV 7 d + GE group).

Article Snippet: HCECs were treated with GE (HY‐N0472, MedChemExpress, USA), the ACE inhibitor MLN‐4760 (100 nM for 24 h) (HY‐19414, MedChemExpress), the Ang‐(1–7) antagonist D‐Pro7‐Ang‐(1–7) (1 μM for 24 h) (50–194‐6749, Thermo Fisher Scientific), the MasR antagonist A‐779 (1 μM for 24 h) (HY‐P0216, MedChemExpress), or neutralising anti‐IL‐10 antibody (2 μg for 24 h) (MAB217, R&D Systems, USA).

Techniques: Staining, Marker