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Qibai Pingfei capsule medicated serum inhibits the proliferation of hypoxia-induced pulmonary arterial smooth muscle cells via the Ca2+/calcineurin/nuclearfactorofactivatedT-cells3pathway

Author: ZhuJie,WangBaoqin,LiuChangan,TongXiangli,LiZegeng

OBJECTIVE: To observe the effect of Qibai Pingfei capsule (QBPF) medicated serum on the proliferation of rat pulmonary arterial smooth muscle cells (PASMCs) under hypoxia conditions and to investigate its key molecular effects on the Ca2+/calcineurin/nuclear factor of activatedT-cells 3 (NFATc3) signalingpathway. METHODS: QBPF was provided to rats via continuous gavage for 10 days. Primary rat PASMCs were cultured using the direct adherent culture method. Methylthiazolyltetrazoliumassaywasusedtoeval uate the effect of QBPF on PASMC proliferation underhypoxiaconditions.Laserscanningconfocalmicroscopy was used to detect changes in intracellular free calcium ([Ca2 + ]i) in PASMC-loaded Fluo-3-AM. Real-time quantitative polymerase chain reaction and western blot were used to detect the transcription and protein expression levels of calcineurin and NFATc3 genesin PASMCs. RESULTS: Compared with normoxia conditions, PASMCs proliferated at 12, 24, 48, and 72 h under hypoxia conditions. QBPF at concentrations of 5%, 10% , and 20% could inhibit hypoxia-induced PASMC proliferation to different degrees.The inhibitory effect was most significant in the 20% QBPF group under 24 h hypoxia conditions. The concentration of [Ca2 +]i in PASMCs under hypoxia was increased and [Ca2 + ]i was significantly decreased when co-incubated with QBPF at 5%, 10%, and 20% . Compared with normoxia conditions, the mRNA and protein expression levels of calcineurin and NFATc3 in PASMCs induced by hypoxia were up-regulated. QBPF application significantly down-regulated mRNA and protein expression levelsofcalcineurinandNFATc3inPASMCs. CONCLUSION: QBPF can effectively inhibit hypoxia-induced proliferation of PASMCs through down-regulation of key molecular expression via theCa2+/calcineurin/NFATc3pathway. Keywords: Hypertension, pulmonary; Myocytes, smooth muscle; Vascular remodeling; NFATC transcription factors

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