瞬时受体电位经典通道1介导血管紧张素Ⅱ诱导的血管平滑肌细胞增殖、迁移及炎症反应OA

Aim To elucidate the role and mecha-nisms of transient receptor potential canonical subtype 1(TRPC1)in angiotensin II(AngII)-induced prolif-eration,migration,and inflammatory factor production of vascular smooth muscle cells(VSMCs).Methods To determine the effect of AngII on TRPC1 expres-sion,VSMCs were cultured and divided into the blank control and AngII groups,using Western blot and RT-qPCR to analyze TRPC1 levels at the protein and mRNA levels,respectively.Vascular smooth muscle-specific TRPC1 knockout mice(TRPC1VSMC-/-)were used to establish an AngII-induced vascular inflamma-tion model.To quantify vascular remodeling,hematoxylin-eosin(HE)staining was used to measure the extent of vascular wall thickening.Cultured VSMCs were seeded and randomly allocated into sev-eral experimental groups,namely,blank control group,siNC+AngII group,and siTRPC1+AngII group.The mRNA expression levels of iNOS,TNF-α,IL-6,IL-1β,and CCL2 were measured by RT-qPCR.Cell proliferation was assessed using the CCK-8 assay,and cell migration was evaluated via a wound healing assay.Intracellular calcium levels mediated by AngII were determined by calcium influx assays and store-operated calcium entry(SOCE)measurements.The protein expression of total MEK,phosphorylated MEK(p-MEK),total ERK1/2,and phosphorylated ERK1/2(p-ERK1/2)was detected by Western blot.Further-more,the MEK/ERK pathway inhibitor PD98059 was applied to investigate whether ERK1/2 phosphorylation mediated the AngII-induced inflammatory response in VSMCs.Results Compared with the blank control group,the mRNA level of TRPC1 in VSMCs treated with AngII was significantly higher than other members of the TRPC family.AngII dose-dependently upregu-lated the protein and mRNA expression levels of TRPC1 in VSMCs.Aortic vascular remodeling in-duced by AngII was ameliorated in TRPC1VSMC-/-mice.Silencing of TRPC1 expression effectively inhibited multiple AngII-induced pro-pathogenic cellular re-sponses.This included the suppression of VSMCs pro-liferation and migration,a reduction in agonist-stimulated calcium influx and SOCE,and the down-regulation of key inflammatory cytokines.TRPC1 par-ticipated in AngII-induced VSMCs proliferation,mi-gration,and inflammatory response through the MEK/ERK1/2 pathway.Conclusion TRPC1 expression is up-regulated in AngII-treated VSMCs.The inhibition of TRPC1 attenuates AngII-induced cell proliferation,migration,and inflammatory response via the MEK/ERK1/2 signaling pathway.