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400037 重庆,陆军军医大学(第三军医大学)第二附属医院泌尿外科
[ 李龙坤,E-mail:lilongk@hotmail.com LI Longkun, E-mail:lilongk@hotmail.com
收稿:2018-10-23,
修回:2019-01-09,
纸质出版:2019-04-15
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张党领, 刘骞, 朱景振, 等. Hcn1通道蛋白表达降低参与大鼠膀胱过度活动症的发生[J]. 第三军医大学学报, 2019,41(7):659-664.
Dangling ZHANG, Qian LIU, Jingzhen ZHU, et al. Downregulation of Hcn1 protein participates in development of overactive bladder in rats[J]. Journal of Third Military Medical University, 2019, 41(7): 659-664.
张党领, 刘骞, 朱景振, 等. Hcn1通道蛋白表达降低参与大鼠膀胱过度活动症的发生[J]. 第三军医大学学报, 2019,41(7):659-664. DOI: 10.16016/j.1000-5404.201810142.
Dangling ZHANG, Qian LIU, Jingzhen ZHU, et al. Downregulation of Hcn1 protein participates in development of overactive bladder in rats[J]. Journal of Third Military Medical University, 2019, 41(7): 659-664. DOI: 10.16016/j.1000-5404.201810142.
目的
2
观察膀胱过度活动症模型(DO)大鼠中核糖核酸(RNA)以及参与的信号通路激活和抑制情况,探讨可能参与膀胱过度活动症发生、发展的信号分子和靶点。
方法
2
膀胱颈结扎法制备DO大鼠模型,尿动力学检测评估模型构建是否成功;常规生物学方法进行RNA提取质控,后期建库测序,数据分析采用R平台完成;q-PCR和Western blot验证测序结论。
结果
2
尿动力学检测显示DO模型构建成功;6例样本的RNA-seq测序结果及后续q-PCR验证DO模型大鼠膀胱组织中6个基因高表达,2个基因低表达(
P
<
0.001);基因组表达富集分析(gene set enr
ichment analysis
GSEA)显示炎症反应相关通路在DO模型大鼠膀胱组织中激活,超极化激活环核苷酸门控阳离子通道(hyperpolarization-activated cyclic nucleotide-gated channel,Hcn)活性被抑制[富集分数=-0.565
错误发现率(FDR)
q
=0.044
]
;Western blot检测结果证实Hcn1通道蛋白在DO模型大鼠中的表达被抑制(
P
<
0.001)。
结论
2
Hcn1通道蛋白表达降低参与大鼠膀胱过度活动症的发生、发展。
Objective
2
To investigate the expression of RNA and activation and inhibition of related signaling pathways in rats with overactive bladder (DO model)
and to explore the molecules and targets involved in the occurrence and development of overactive bladder.
Methods
2
Rat DO model was prepared by bladder neck ligation. The urodynamic test was used to evaluate the success of establishment. The regular biological methods were used for RNA extract and quality control was completed by corporation. The data analysis was performed by R platform. Q-PCR and Western blotting were employed to confirm the results of sequencing.
Results
2
The result of urodynamic test showed that the DO model was successfully constructed. The RNA-seq sequencing results from 6 samples and the subsequent RT-PCR results showed that the 6 genes in DO bladder tissue were highly expressed and 2 were lowly expressed (
P
<
0.001). Gene set enrichment analysis (GSEA) showed that the inflammatory response-related pathway was activated in the DO model
and voltage-gated sodium channel activity was inhibited (Enrichment score=-0.565
FDR
q
value=0.044); Western blot analysis confirmed that hyperpolarization-activated cyclic nucleotide-gated channel 1 (Hcn1) channel protein in DO rats was down-expressed (
P
<
0.001).
Conclusion
2
Low expression of Hcn1 channel protein is involved in the occurrence and development of overactive bladder in the rats.
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