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Acid sphingomyelinase/ceramide regulates carotid intima-media thickness in simulated weightless rats

  • Molecular and Cellular Mechanisms of Disease
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Abstract

Structural adaptation of arteries to weightlessness might lower the working ability or even threaten the physical health of astronauts, but the underlying mechanism is unclear. Acid sphingomyelinase (ASM) catalyzes ceramide (Cer) generation controlling arterial remodeling through multiple signaling pathways. In the present study, we aimed to investigate the contribution of ASM/Cer to the changes of common carotid artery intima-media thickness (CIMT) induced by simulated weightlessness. Hindlimb-unloaded tail-suspended (HU) rats were used to simulate the effect of weightlessness. Morphology of the carotid artery (CA) was examined by hematoxylin-eosin staining. Protein content of ASM or proliferating cell nuclear antigen (PCNA) was detected by Western blot. Cer level was measured by immunohistochemistry analysis. Apoptosis events were observed by transferase-mediated dUTP nick end labeling (TUNEL) staining. During 4 weeks of tail suspension, CIMT was increased gradually in HU but not in their synchronous control rats (P < 0.05). Correspondingly, the CA of HU rats had a lower apoptosis and higher proliferation of vascular smooth muscle cells (VSMCs). As compared to the control, both ASM protein expression and Cer content were reduced significantly in CA of HU rats (P < 0.05), incubation of which with permeable Cer reversed the changes in apoptosis and proliferation substantially. Furthermore, when the ASM protein content as well as Cer level in CA of control rats was diminished by using an ASM inhibitor, an increase of CIMT along with reduced apoptosis and enhanced proliferation of VSMCs was found. Our results suggest that by controlling the balance between apoptosis and proliferation, ASM/Cer plays an important role in the regulation of CIMT during simulated weightlessness.

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Acknowledgement

This study was supported by the National Natural Science Foundation of China (Grant No. 31071045, 81401550, 81671856).

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  1. Department of Aerospace Hygiene, Fourth Military Medical University, Xi’an, 710032, People’s Republic of China

    Yao-Ping Cheng, Hai-Jun Zhang, Yu-Ting Su, Xing-Xing Meng, Xiao-Ping Xie, Yao-Ming Chang & Jun-Xiang Bao

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  1. Yao-Ping Cheng
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Correspondence to Jun-Xiang Bao.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The protocols and procedures performed in the studies involving animals were in accordance with the ethical standards of the Fourth Military Medical University of P.R. China.

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Yao-Ping Cheng and Hai-Jun Zhang contributed equally to the work.

Electronic supplementary material

Supplementary Fig. 1

Change of Cer content after different time of C6-Cer incubation in CA of CON and HU rats. a and b, original picture (a) and summarized data (b) showing the abundance of Cer in CA of rats with C6-Cer incubation for 0 h, 1 h, 2 h and 4 h detected by immunohistofluorescence assay. Magnification ×600; Scale bar, 50 μm; L, lumen. One-way ANOVA and student t-test were used to summarize the data. Values are means ± SEM. n = 4, * P < 0.05 vs. synchronous CON, # P < 0.05 vs. HU 0 h (GIF 58 kb)

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Supplementary Fig. 2

Change of bax/bcl-2 expression in CA after different time of hindlimb unloading tail suspension in rats. a and b, representative band (a) and summarized data (b) showing bax/bcl-2 ratio in CA of rats with HU treatment for 0d, 3d, 1w, 2w and 4w detected by Western blot. One-way ANOVA and Dunnett-t test were used to summarize the data. Values are means ± SEM. n = 4, * P < 0.05 vs. 0d (GIF 41 kb)

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Cheng, YP., Zhang, HJ., Su, YT. et al. Acid sphingomyelinase/ceramide regulates carotid intima-media thickness in simulated weightless rats. Pflugers Arch - Eur J Physiol 469, 751–765 (2017). https://doi.org/10.1007/s00424-017-1969-z

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