GlcNAc经O-糖基化依赖性途径抑制大鼠阴茎海绵体平滑肌细胞中MAPKs通路

刘博深; 王轶; 郭林民; 孙中义

doi:10.16016/j.1000-5404.201808180

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GlcNAc经O-糖基化依赖性途径抑制大鼠阴茎海绵体平滑肌细胞中MAPKs通路

基础医学 | 更新时间：2026-02-12

- GlcNAc经O-糖基化依赖性途径抑制大鼠阴茎海绵体平滑肌细胞中MAPKs通路
- GlcNAc inhibits MAPKs pathway in rat corpus cavernosum smooth muscle cells via O-glycosylation-dependent pathway
- 第三军医大学学报 2019年41卷第3期页码：206-212
- 作者机构：
  
  400042 重庆，陆军军医大学(第三军医大学)第三附属医院(野战外科研究所)泌尿外科Department of Urology, Institute of Surgery Research, Third Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042, China
- 作者简介：
- 基金信息：
  
  国家国际科技合作专项项目(2014DFR30860);陆军军医大学科技成果转化基金项目(2016XZH16);重庆高校创新团队建设计划资助项目(2016)
- DOI：10.16016/j.1000-5404.201808180
  中图分类号： R-33;R322.64;R329.26
- 收稿：2018-08-29，
  
  纸质出版：2019-02-15
- 稿件说明：
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刘博深, 王轶, 郭林民, 等. GlcNAc经O-糖基化依赖性途径抑制大鼠阴茎海绵体平滑肌细胞中MAPKs通路[J]. 第三军医大学学报, 2019,41(3):206-212.

Boshen LIU, Yi WANG, Linmin GUO, et al. GlcNAc inhibits MAPKs pathway in rat corpus cavernosum smooth muscle cells via O-glycosylation-dependent pathway[J]. Journal of Third Military Medical University, 2019, 41(3): 206-212.
刘博深, 王轶, 郭林民, 等. GlcNAc经O-糖基化依赖性途径抑制大鼠阴茎海绵体平滑肌细胞中MAPKs通路[J]. 第三军医大学学报, 2019,41(3):206-212. DOI： 10.16016/j.1000-5404.201808180.

Boshen LIU, Yi WANG, Linmin GUO, et al. GlcNAc inhibits MAPKs pathway in rat corpus cavernosum smooth muscle cells via O-glycosylation-dependent pathway[J]. Journal of Third Military Medical University, 2019, 41(3): 206-212. DOI： 10.16016/j.1000-5404.201808180.

摘要

目的

探讨β-N-乙酰氨基葡萄糖(β-N-acetylglucosamine

GlcNAc)对大鼠阴茎海绵体平滑肌细胞促分裂原活化蛋白激酶(mitogen-activated protein kinases

MAPKs)通路的影响。

方法

分离健康成年大鼠阴茎组织，高糖培养基诱导阴茎海绵体平滑肌细胞(cavernous smooth muscle cells

CSMC)分离生长，免疫荧光、免疫组化鉴定CSMC，GlcNAc处理CSMC 12 h，分别于0、10 min、1、2、4、8、12 h收集细胞裂解液，O-(连接)N-乙酰葡糖胺转移酶(O-N-acetylglucosamine transferase

OGT)抑制剂Alloxan、O-(连接)N-乙酰葡糖胺水解酶(O-GlcNAcase

OGA)抑制剂Thiamet G及GlcNAc处理CSMC 72 h

Western blot检测蛋白总糖基化水平及MAPKs通路相关蛋白在12、72 h的磷酸化水平。

结果

成功分离培养大鼠CSMC，GlcNAc显著提升了CSMC细胞O-糖基化水平(

0.05)。处理细胞后12 h，GlcNAc显著抑制大鼠CSMC的ERK1/2、JNK1/2/3和p38的蛋白磷酸化水平(

0.05)，处理细胞后72 h，GlcNAc显著抑制了大鼠CSMC的ERK1/2和p38蛋白磷酸化水平(

0.05)，但不影响其总蛋白的表达水平(

0.05)，且与Thiamet G的作用相同。另外，GlcNAc与Thiamet G能协同抑制ERK1/2、JNK1/2/3和p38的蛋白磷酸化水平(

0.05)。

结论

GlcNAc经O-糖基化依赖性途径抑制阴茎海绵体平滑肌细胞中MAPKs通路。

Abstract

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

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 (

0.05). At 12 h

GlcNAc sig

nificantly inhibited ERK1/2

JNK1/2/3 and p38 (

0.05)

while at 72 h

GlcNAc significantly inhibited the phosphorylation of ERK1/2 and p38 protein in rat CSMC (

0.05)

but did not affect the levels of total proteins (

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 (

0.05).

Conclusion

GlcNAc inhibits MAPKs pathway in CSMC

via

O-glycosylation-dependent pathway.

关键词

Keywords

references

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