GlcNAc inhibits MAPKs pathway in rat corpus cavernosum smooth muscle cells via O-glycosylation-dependent pathway

Di-san junyi daxue xuebao

SUN Zhongyi¹  GUO Linmin¹  WANG Yi¹  LIU Boshen¹ 

[1] Department of Urology, Institute of Surgery Research, Third Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042, China ;

关键词: cavernous smooth muscle cells;    β-n-acetylglucosamine;    o-glycosylation;    mitogen-activated protein kinases;   

DOI  :  10.16016/j.1000-5404.201808180

来源: DOAJ

【 摘 要 】

Objective To investigate the effect of β-N-acetylglucosamine (GlcNAc) on the mitogen-activated protein kinases (MAPKs) pathway in rat penile cavernosum smooth muscle cells. Methods After penile tissue was collected from healthy adult rats, cavernous smooth muscle cells (CSMC) were isolated and cultured under high glucose medium. Immunofluorescence (IF) assay and immunocytochemical (ICC) assay were used to identify CSMC. Then the cells were treated with GlcNAc for 12 h, the cell lysate was collected at 0, 10 min, and 1, 2, 4, 8 and 12 h. Western blotting was employed to detect the total glycosylation level of protein and phosphorylation of MAPKs pathway-associated proteins after the cells were treated with O-N-acetylglucosamine transferase (OGT) inhibitor, Alloxan, O-GlcNAcase (OGA) inhibitor, Thiamet G, and GlcNAc. Results The rat CMSC were successfully isolated. Compared with the control cells, GlcNAc treatment significantly increased the O-glycosylation level of CSMC (P < 0.05). At 12 h, GlcNAc significantly inhibited ERK1/2, JNK1/2/3 and p38 (P < 0.05), while at 72 h, GlcNAc significantly inhibited the phosphorylation of ERK1/2 and p38 protein in rat CSMC (P < 0.05), but did not affect the levels of total proteins (P>0.05), which was the same effect of Thiamet G. In addition, GlcNAc and Thiamet G synergistically inhibited the phosphorylation levels of ERK1/2, JNK1/2/3 and p38 (P < 0.05). Conclusion GlcNAc inhibits MAPKs pathway in CSMC via O-glycosylation-dependent pathway.

【 授权许可】

Unknown