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. 2019 Jul 11;12:5499–5513. doi: 10.2147/OTT.S209291

Figure 7.

Figure 7

Down-regulation of GRK2 in M2-polarized macrophages activated the β2-AR downstream signaling pathway. (A) An ELISA was performed to determine the cAMP level in M2-polarized macrophages stimulated with Terb for 5, 10 or 15 min after the transfection with GRK2 siRNA. In the Terb-alone group, the formation of cAMP peaked at 5 min, and declined from 5 min to 15 min. While in GRK2 siRNA+Terb group, there was a significant elevation in cAMP formation and the peak time was delayed to 10 min. Furthermore, at 15 min, the level of cAMP in the GRK2 siRNA+Terb group was higher compared with the Terb-alone group. *P<0.05 and **P<0.01 compared with the control group; #P<0.05 and ##P<0.01 compared with each other group at the same time point. (B) Western blot analysis was performed to determine the level of PKA in M2-polarized macrophages stimulated with Terb for 10 min after transfection with GRK2 siRNA. The expression of PKA was increased in the GRK2 siRNA+Terb group compared with Terb-alone group. (C)Western blot analysis was performed to determine the expression levels of CREB, p-CREB, STAT3, p-STAT3 levels in M2-polarized macrophages stimulated with Terb for 10 min after the transfection with GRK2 siRNA. No significant elevation was observed in the expression of CREB and STAT3 in the GRK2 siRNA+Terb group compared with the Terb-alone group, while p-CREB and p-STAT3 levels were significantly increased. *P<0.05 and **P<0.01 compared with the control group; #P<0.05 and ##P<0.01 compared with the Terb group.