Proliferation and activation of vascular smooth muscle cells induced by activated complement alternative pathway and intervention study / 中国药理学通报

ABSTRACT

Aim To study the effect of proliferation and activation of vascular smooth muscle cells (VSMCs) induced by activated the complement alternative pathway and intervention. Methods Normal human plasma was specifically activated the complement alternative pathway by incubated with cobra venom factor (CVF). Exposed VSMCs to the activated complement products, the change of cell morphology and the cell viability were assayed by inverted phase contrast microscope and MTT method, respectively. The supernatant was assayed for expression of E-selectin, ICAM-1 and VCAM-1 by using ELISA reagent kits. The protein expression levels of p-NF-kB p65, NF-kB p65 and IKK were assayed by Western blot. The nucleus transcriptional activity of NF-ΚB p65 was tested by the dual luciferase reporter assay system. Pyrrolidine dithiocarbamate (PDTC) was used to intervene the proliferation and activation of VSMCs. Results VSMCs were activated and induced to proliferation after exposed to the products of activated complement alternative pathway. The expressions of E-selectin, VCAM-1 and ICAM-1 were up-regulated. The contents of ICAM-1 and VCAM-1 reached the peak at 6 h and the E-selectin increased significantly at 12 h. Meanwhile, the phosphorylation level of NF-ΚB p65, nucleus transcriptional activity of NF-ΚB p65 and the protein expression of IKK and N F - Κ B p65 increased. The above mentioned changes were clearly inhibited by PDTC. Conclusions The activated complement alternative pathway can initiate proliferation and activation of VSMCs, and its mechanism goes hand in hand with activation of N F - Κ B signaling pathway. PDTC, a specific inhibitor of N F - K B, can effectively inhibit the proliferation and activation of VSMCs.