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FIGURE 2 | QSG inhibited activation of TGF-β1/Smad3 through reducing the recruitment of M1 macrophages in myocardial tissue. (A) IF staining showed that splenectomy reduced the recruitment of M1 macrophages in myocardial tissue; QSG treatment decreased the recruitment of M1 macrophages. (B) Masson and Sirius Red staining showed that QSG inhibited cardiac fibrosis. (C) Western blot showed that QSG down-regulated the expressions of AT1, MCP-1 and <t>CCR2</t> in myocardial tissue compared with that in the model group. (D) Western blot showed that QSG decreased the expressions of TGF-β1, Smad3, collagen III and MMP2 in cardiac tissue. All data were presented as means ± SD from independent experiments performed in triplicate. #P < 0.05, ##P < 0.01, ###P < 0.001 vs the sham group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the model group. N = 3 per group.
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FIGURE 2 | QSG inhibited activation of TGF-β1/Smad3 through reducing the recruitment of M1 macrophages in myocardial tissue. (A) IF staining showed that splenectomy reduced the recruitment of M1 macrophages in myocardial tissue; QSG treatment decreased the recruitment of M1 macrophages. (B) Masson and Sirius Red staining showed that QSG inhibited cardiac fibrosis. (C) Western blot showed that QSG down-regulated the expressions of AT1, MCP-1 and <t>CCR2</t> in myocardial tissue compared with that in the model group. (D) Western blot showed that QSG decreased the expressions of TGF-β1, Smad3, collagen III and MMP2 in cardiac tissue. All data were presented as means ± SD from independent experiments performed in triplicate. #P < 0.05, ##P < 0.01, ###P < 0.001 vs the sham group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the model group. N = 3 per group.
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FIGURE 2 | QSG inhibited activation of TGF-β1/Smad3 through reducing the recruitment of M1 macrophages in myocardial tissue. (A) IF staining showed that splenectomy reduced the recruitment of M1 macrophages in myocardial tissue; QSG treatment decreased the recruitment of M1 macrophages. (B) Masson and Sirius Red staining showed that QSG inhibited cardiac fibrosis. (C) Western blot showed that QSG down-regulated the expressions of AT1, MCP-1 and <t>CCR2</t> in myocardial tissue compared with that in the model group. (D) Western blot showed that QSG decreased the expressions of TGF-β1, Smad3, collagen III and MMP2 in cardiac tissue. All data were presented as means ± SD from independent experiments performed in triplicate. #P < 0.05, ##P < 0.01, ###P < 0.001 vs the sham group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the model group. N = 3 per group.
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FIGURE 2 | QSG inhibited activation of TGF-β1/Smad3 through reducing the recruitment of M1 macrophages in myocardial tissue. (A) IF staining showed that splenectomy reduced the recruitment of M1 macrophages in myocardial tissue; QSG treatment decreased the recruitment of M1 macrophages. (B) Masson and Sirius Red staining showed that QSG inhibited cardiac fibrosis. (C) Western blot showed that QSG down-regulated the expressions of AT1, MCP-1 and CCR2 in myocardial tissue compared with that in the model group. (D) Western blot showed that QSG decreased the expressions of TGF-β1, Smad3, collagen III and MMP2 in cardiac tissue. All data were presented as means ± SD from independent experiments performed in triplicate. #P < 0.05, ##P < 0.01, ###P < 0.001 vs the sham group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the model group. N = 3 per group.

Journal: Frontiers in pharmacology

Article Title: Qishen Granule Improved Cardiac Remodeling via Balancing M1 and M2 Macrophages.

doi: 10.3389/fphar.2019.01399

Figure Lengend Snippet: FIGURE 2 | QSG inhibited activation of TGF-β1/Smad3 through reducing the recruitment of M1 macrophages in myocardial tissue. (A) IF staining showed that splenectomy reduced the recruitment of M1 macrophages in myocardial tissue; QSG treatment decreased the recruitment of M1 macrophages. (B) Masson and Sirius Red staining showed that QSG inhibited cardiac fibrosis. (C) Western blot showed that QSG down-regulated the expressions of AT1, MCP-1 and CCR2 in myocardial tissue compared with that in the model group. (D) Western blot showed that QSG decreased the expressions of TGF-β1, Smad3, collagen III and MMP2 in cardiac tissue. All data were presented as means ± SD from independent experiments performed in triplicate. #P < 0.05, ##P < 0.01, ###P < 0.001 vs the sham group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the model group. N = 3 per group.

Article Snippet: After electrophoresis at 100 V for 1–1.5 h, proteins were transferred to PVDF membranes at 300 mA for 1–1.5 h. Afterwards, the membrane blocked with 5% skim milk for 1–1.5 h at room temperature, incubated on a shaker, and washed with TBS-T. Western blot analysis was conducted using anti-AT1 (ab18801; Abcam, United States), Frontiers in Pharmacology | www.frontiersin.org November 2019 | Volume 10 | Article 1399 anti-MCP-1 (ab25124, Abcam, United States), anti-CCR2 (PAI-27409), anti-TGF-β1 (3711s, Cell Signaling Technology, Germany), anti-Smad3 (ab28379, Abcam, United States), antiMMP2 (ab86607, Abcam, United States), anti-Col III (ab7778, Abcam, United States), anti-VEGF (ab10972, Abcam, United States), anti-CD31 (ab24590, Abcam, United States) and antiGAPDH (ab8245, Abcam, United States) at 4°C overnight.

Techniques: Activation Assay, Staining, Western Blot

FIGURE 5 | QSG suppresses the release of spleen monocytes and recruitment to heart tissues through the splenic Ang II/AT1-cardiac MCP-1/CCR2 pathway to inhibit MR.

Journal: Frontiers in pharmacology

Article Title: Qishen Granule Improved Cardiac Remodeling via Balancing M1 and M2 Macrophages.

doi: 10.3389/fphar.2019.01399

Figure Lengend Snippet: FIGURE 5 | QSG suppresses the release of spleen monocytes and recruitment to heart tissues through the splenic Ang II/AT1-cardiac MCP-1/CCR2 pathway to inhibit MR.

Article Snippet: After electrophoresis at 100 V for 1–1.5 h, proteins were transferred to PVDF membranes at 300 mA for 1–1.5 h. Afterwards, the membrane blocked with 5% skim milk for 1–1.5 h at room temperature, incubated on a shaker, and washed with TBS-T. Western blot analysis was conducted using anti-AT1 (ab18801; Abcam, United States), Frontiers in Pharmacology | www.frontiersin.org November 2019 | Volume 10 | Article 1399 anti-MCP-1 (ab25124, Abcam, United States), anti-CCR2 (PAI-27409), anti-TGF-β1 (3711s, Cell Signaling Technology, Germany), anti-Smad3 (ab28379, Abcam, United States), antiMMP2 (ab86607, Abcam, United States), anti-Col III (ab7778, Abcam, United States), anti-VEGF (ab10972, Abcam, United States), anti-CD31 (ab24590, Abcam, United States) and antiGAPDH (ab8245, Abcam, United States) at 4°C overnight.

Techniques: