Silencing of STIM1 attenuates hypoxia-induced PASMCs proliferation via inhibition of the SOC/Ca2+/NFAT pathway

Respiratory Research

Yuqi Gao¹  Gang Xu²  Fuyu Liu²  Yongjun Luo²  Jian Chen²  Xianhua Hou² 

[1] Department of Pathophysiology and high altitude physiology, College of high altitude military medicine, Third Military Medical University, Chongqing, China;Key Laboratory of High Altitude Medicine of PLA, 400038, Chongqing, China

关键词: Cell proliferation;    Hypoxia;    Pulmonary hypertension;    RNA interference;    Stromal interaction molecule 1;   

Others  :  796564
DOI  :  10.1186/1465-9921-14-2

 received in 2012-03-01, accepted in 2012-12-19,  发布年份 2013

【 摘 要 】

Background

Stromal interaction molecule 1 (STIM1) is a newly discovered Ca²⁺ sensor on the endoplasmic reticulum which is an indispensable part in the activation of store-operated Ca²⁺ channels (SOC). Recent studies demonstrate that SOC of pulmonary smooth muscle cells (PASMCs) were upregulated by chronic hypoxia which contribute to the enhanced pulmonary vasoconstriction and vascular remodeling. However, the exact role of STIM1 in the development of chronic hypoxic pulmonary hypertension(HPH) remains unclear.

Methods

In this study we investigated the cellular distribution and expression of STIM1 by immunofluorescence, qRTPCR and Western blotting methods in Wistar rat distal intrapulmonary arteries under normal and chronic hypobaric hypoxic conditions. In vitro, Wistar rat PASMCs were isolated and cultured. PASMCs were transfected with siRNA targeting STIM1 gene by liposome. The expression of STIM1 protein was detected by Western blotting. [³H]-thymidine ([³H]-TdR) incorporation were performed to detect PASMCs proliferation. The cell cycle was analyzed by flow cytometry. The SOC-mediated Ca²⁺ influx was calculated by Ca²⁺ fluorescence imaging and the nuclear translocation of NFATc3 was determined by immunofluorescence and Western blot analysis of nuclear extracts.

Results

We found that during the development of HPH and the initiation of vascular remodeling, the mRNA and protein expression levels of STIM1 significantly increased in the distal intrapulmonary arteries. Moderate hypoxia significantly promotes PASMCs proliferation and cell cycle progression. Silencing of STIM1 significantly decreased cellular proliferation and delayed the cell cycle progression induced by hypoxia. Silencing of STIM1 also significantly decreased SOC-mediated Ca²⁺ influx and inhibited the nuclear translocation of NFATc3 in hypoxic PASMCs.

Conclusion

Our findings suggest that chronic hypobaric hypoxia upregulates the expression of STIM1 in the distal intrapulmonary arteries which plays an important role in the hypoxia-induced PASMCs proliferation via SOC/Ca²⁺/NFAT pathway and may represent a novel therapeutic target for the prevention of hypoxia pulmonary hypertension.

【 授权许可】

   
2013 Hou et al; licensee BioMed Central Ltd.

【 图 表 】

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