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Abstract

Volume 12, Issue 5 (September 2010) 12, 744–752; 10.1038/aja.2010.44

Effect of tankyrase 1 on autophagy in the corpus cavernosum smooth muscle cells from ageing rats with erectile dysfunction and its potential mechanism

Jian Zhang1, Xiao-Jun Wu2, De-Xiang Zhuo3, Tao Liu1, Wei-Ren Li1, Ze-Bin Mao3, Zhong-Cheng Xin1

1 Andrology Center, First Hospital, Peking University, Beijing 100009, China
2 Institute of Urinary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
3 The Department of Biochemistry and Molecular Biology, Health Science Center, Peking University, Beijing 100191, China

Correspondence: Dr Zhong-Cheng Xin,xinzc@bjmu.edu.cn

Received 16 March 2010; Revised 19 April 2010; Accepted 7 May 2010; Published online 26 July 2010.

Abstract

This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction (ED) and non-ED rats' corpus cavernosum smooth muscle cells (CSMCs). This study aslo explored the effect and possible mechanism of tankyrase 1 on autophagy and cell proliferation in ageing ED rats' CSMCs. The intracavernous pressure and mean systemic arterial pressure were measured to investigate erectile function so that eight 24-month-old ED and eight 8-month-old male Wistar rats were choosed respectively. The rat CSMCs were isolated and cultured by enzyme digestion, in which tankyrase 1 expression and autophagy quantity were compared. Tankyrase 1 overexpression was induced with plasmid transfection by Lipofectamine™. The effect of tankyrase 1 overexpression on proliferation, autophagy and mTOR pathway in 24-month-old ED rats' CSMCs was measured by the cell growth curve in MTT assay, cell cycle analysis in flow cytometry (FCM), key protein expression in Western blot, autophagy quantity in transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 fluorescence. The primary CSMCs were confirmed by immunofluorescence, and the purity was 99.1% in FCM. Compared with that of 8-month-old rats, tankyrase 1 expression and autophagy quantity significantly decreased in 24-month-old ED rats' primary CSMCs (P < 0.01). Tankyrase 1 overexpression significantly increased the growth rate (P < 0.05) and increased the S phase of cell cycle (P < 0.01). The autophagosome quantity was remarkably increased (P < 0.01), LC3-I/II and Beclin 1 were upregulated (P < 0.01 and P < 0.05), and p-p70S6K (Thr389) was downregulated in 24-month-old ED rat CSMCs (P < 0.05). In conclusion, Tankyrase 1 and autophagy decrease in the CSMCs from aging rats with ED, and tankyrase 1 may have a positive effect on proliferation by enhancing autophagy and regulating the mTOR signalling pathway.


Keywords:

ageing; autophagy; corpora cavernosum; corpus cavernosum smooth muscle cells; erectile dysfunction; senile; tankyrase

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