FIGURE 4.

PIP₃/Akt signaling regulated the production of Col8a1. (A) IPA software was used to analyze related molecules that can participate in the regulation of “formation of extracellular matrix” and (B) the core signaling pathways in the cytoplasm and nucleus participated in this network. (C) Western blotting was used to measure the expression of phosphorylated Akt in VSMCs, and the results showed that Akt was significantly activated after pEV stimulation. (D–F) ELISA and western blotting were used to analyze molecules involved in the activation of Akt. Only the level of PIP₃ increased significantly in VSMCs after pEV stimulation, (D) while PDK1 (E) and mTORC (F) were unchanged. (G–J) The specific PIP₃/Akt signaling inhibitor LY294002 was used to examine the effect of PIP₃/Akt signaling on the production of Col8a1. (G) LY294002 significantly abrogated the expression of p-Akt induced by pEVs and blocked the mRNA (I) and protein expression of col8a1 (H,J). The values are shown as the mean ± SD, *P < 0.05, **P < 0.01 vs. control (n = 4 biological replicates).