Figure 3

Ibandronate decreases proliferation, CTGF protein expression, Hyp upregulation elicited by NE in cultured VSMCs from SHR. (A) Quiescent VSMCs from both strains were pre-incubated with ibandronate (5, 10 μM) for 2 h and then incubated with or without NE (1 μM) for 24 h. Cell proliferation was expressed as the optical density which was determined by Cell Counting Kit (CCK-8). (B) Quiescent VSMCs from both strains were pre-incubated with ibandronate (5, 10 μM) for 2 h and then incubated with or without NE (0.1 μM) for 24 h. Hydroxyproline (Hyp) levels (as an index of collagen) in the culture supernatant were measured by the Hyp test kit. (C) Quiescent VSMCs from both strains were pre-incubated with ibandronate (5, 10 μM) for 2 h and then incubated with or without NE (0.1 μM) for 24 h. Levels of CTGF protein was determined by Western blot. The ratio of CTGF is set to ‘1’ for WKY control group. (D) Effects of ibandronate on the apoptosis of SHR-VSMCs. Incubation of quiescent SHR-VSMCs with ibandronate in a series of final concentrations (0, 5, and 10 μM) for 24 h induced no apoptosis. Representative diagraphs altered by different concentrations of Ibandronate are shown: (a) control, (b) treatment with 5 μM of ibandronate, (c) treatment with 10 μM of ibandronate, (d) representative bar chart showing percentage of apoptotic cells. Data are expressed as mean±s.e.m. (n=5). ^#P<0.05, ^##P<0.01 vs corresponding WKY group; *P<0.05, ^**P<0.01 vs corresponding control group. ^aP<0.05, ^aaP<0.01 vs NE treated SHR-VSMC group. A full color version of this figure is available at the Hypertension Research journal online.