ErMiao San Inhibits Angiogenesis in Rheumatoid Arthritis by Suppressing JAK/STAT Signaling Pathways

Figure 4

EMS inhibits VEGF-induced HUVECs migration, invasion, and tube formation. (a, e) HUVECs were placed in 48-well culture plates and wounded by scratching with pipette tips. Then they are allowed to migrate for 12 h in the presence of VEGF (20 ng/mL) with or without EMS (0.2, 0.4, and 0.8 mg/mL). The cell migration distance was calculated. (b, f) HUVECs were seeded in upper chamber with serum-free H-DMEM while VEGF were put in lower chamber with 10%FBS H-DMEM. It is allowed to migrate for 6 h in the presence of EMS (0.2, 0.4, and 0.8 mg/mL) or not, then the migration cells were counted. (c, g) HUVECs were seeded in matrigel precoated upper chamber with serum-free H-DMEM. It is allowed to migrate for 14 h in the presence of VEGF (20 ng/mL) with or without EMS (0.2, 0.4, and 0.8 mg/mL), then the invasion numbers were counted. (d, h) HUVECs were plated on the matrigel precoated 48-well plates with or without the presence of VEGF (20 ng/mL) and/or EMS for 6 h. Quantitation of the antiangiogenic activity of EMS on tube formation was by counting the number of branch points. (i) HUVECs were plated on the 96-well plates with or without the presence of VEGF (20 ng/mL) and/or EMS for 24 h and tested by MTT. All experiments were done in triplicate. Mean ± SEM (n = 3) was calculated from independent experiments. , compared to the control group; , , and , compared to the VEGF-induced group.

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