Effects of endothelin-1 on monocyte chemotactic protein-1 production in rat vascular smooth muscle cells and its mechanism

doi:10.12047/j.cjap.5611.2018.066

Abstract

Abstract: Objective: To observe the effects of endothelin-1 (ET-1) on monocyte chemotactic protein-1 (MCP-1) generation and the primary mechanisms in rat vascular smooth muscle cells (VSMCs).Methods: Rats VSMCs were cultured and divided into two groups:ET-1 group and inhibitor group (the latter including BQ123+ET-1, BQ788+ET-1, NAC+ET-1, PD98059+ET-1, SB203580+ET-1, SP600125+ET-1 group). ①The ET-1 group was stimulated with different concentration of ET-1 for indicated time. The VSMCs in inhibitor groups were incubated with various inhibitor, including BQ123 and BQ788 (ET_A and ET_B receptor antagonist), antioxidant NAC (N-acetyl cysteine), PD98059 (ERK inhibitor), SB203580 (p38MAPK inhibitor), SP600125 (JNK inhibitor), and PDTC (NF-κB inhibitor) for 30 minutes, then incubated with ET-1 for 24 hours. At indicated time, concentration of MCP-1 protein and expression of MCP-1 mRNA were determined by ELISA and RT-PCR respectively. ②The VSMCs were incubated with inhibitors (including BQ123, BQ788, NAC, PD98059, SB203580, and SP600125 for 20 minutes, then incubated with ET-1 for 5 minutes. The levels of ERK, p38MAPK, JNK, and corresponding phosphorylated protein of them (i.e. p-ERK, p-p38MAPK, p-JNK) were determined by Western blot. Each assay was repeated by three independent experiments.Results: The results showed that ET-1 could stimulate MCP-1 generation in VSMCs both in protein and in mRNA levels in concentration-dependence manner(P<0.05, P<0.01). BQ123, NAC, PD98059, SB203580 and PDTC, but not BQ788 and SP600125, could inhibite ET-1-stimulated protein and mRNA expressions of MCP-1 in VSMCs (P<0.01). In addition, BQ123, NAC, and PD98059 or SB203580 could suppress ET-1 induced ERK and p38MAPK activation respectively (P<0.05, P<0.01), but there was no increase of phospho-JNK in ET-1 treated VSMCs.Conclusion: The results demonstrate that ET-1 induces MCP-1 production in VSMCs via ET_A receptor and subsequent ROS, ERK, p38MAPK, NF-κB signal pathway.

Key words: endothelin-1, monocyte chemotactic protein-1, inflammation, atherosclerosis, vascular smooth muscle cells, rat

WANG Chen-jing, PEI Shu-yan, NAN Xiao-dong, MA Yan-qing. Effects of endothelin-1 on monocyte chemotactic protein-1 production in rat vascular smooth muscle cells and its mechanism[J]. CJAP, 2018, 34(3): 283-288.

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