HBMECs were cultured for 24 h, and RASMCs were cultured for 48 h, then washed twice with ice cold PBS on ice and lysed in NP40 lysis buffer (Biosource, Camarillo, CA, USA) (50 mM Tris, pH 7.
RASMCs were incubated with astaxanthin (1-10 [micro]M) for 48 h robustly leading to a rapid decrease in p-GSK3[beta] and increase wnt5a, [beta]-catenin and cyclin D1 expression, as shown in Fig.
Similarly, the use of IWR-1-endo increased the expression of p-GSK3[beta], reduced the expression of wnt7a, [beta]-catenin, cyclin D1 in RASMCs, as shown in Fig.
The major finding of the present study is that astaxanthin induce proliferation, migration and tube formation in HBMECs and RASMCs in vitro.
When different concentrations of astaxanthin cultivate HBMECs and RASMCs, the expression of p-GSK3[beta] decreased obviously, while [beta]-catenin expression increased significantly.
In our study, astaxanthin up-regulated [beta]-catenin expression in HBMECs and RASMCs, the major effects of astaxanthin appeared to take place via anti-phosphorylation rather than absolute alterations of protein levels.
Taken together, our findings suggest that astaxanthin may be a novel way to induce angiogenesis and play synergistic effect in HBMECs and RASMCs.