[1]熊智勇,周鹏,谢秋波,等.RNA干扰HSP27基因表达对膀胱逼尿肌细胞收缩特性的影响[J].第三军医大学学报,2015,37(06):500-504.
 Xiong Zhiyong,Zhou Peng,Xie Qiubo,et al.Effect of RNA interference of heat shock protein 27 expression on contraction function of human detrusor cells in vitro[J].J Third Mil Med Univ,2015,37(06):500-504.
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RNA干扰HSP27基因表达对膀胱逼尿肌细胞收缩特性的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第06期
页码:
500-504
栏目:
专题报道
出版日期:
2015-03-30

文章信息/Info

Title:
Effect of RNA interference of heat shock protein 27 expression on contraction function of human detrusor cells in vitro
作者:
熊智勇周鹏谢秋波卢根生王永权
第三军医大学西南医院全军泌尿外科研究所;中国人民解放军95333部队
Author(s):
Xiong Zhiyong Zhou Peng Xie Qiubo Lu Gensheng Wang Yongquan

Institute of Urology Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400038; Troop 95333 of PLA, Changsha, Hunan Province, 410000, China

关键词:
热休克蛋白27肌动蛋白逼尿肌细胞膀胱收缩功能障碍
Keywords:
HSP27 actin detrusor cells Bladder contraction dysfunction
分类号:
R322.62;R341;R394.2
文献标志码:
A
摘要:

目的      探讨热休克蛋白27(heat shock protein 27, HSP27)低表达对体外培养人膀胱逼尿肌细胞(bladder smooth muscle cells,BSMCs)肌动蛋白骨架结构及其收缩功能的影响。      方法      构建HSP27基因干扰慢病毒载体作用于BSMCs后,Western blot检测其抑制HSP27表达水平和干扰效率;通过鬼笔环肽荧光染色、Western blot检测BSMCs内F/G-actin比值检测等方法观察,干扰HSP27表达对BSMCs肌动蛋白骨架形态结构和聚合状态的影响;构建离体BSMCs三维培养模型,检测HSP27低表达对膀胱逼尿肌细胞收缩力的影响。      结果      成功构建HSP27干扰质粒pGreenPuro-sh-HSP27,慢病毒载体高效感染BSMCs;与正常对照组比较,HSP27干扰组BSMCs细胞内肌动蛋白骨架结构紊乱和皱缩,检测F/G-actin比值(0.116±0.001)亦较正常BSMCs (8.717±0.500)明显下降(P<0.05);正常BSMCs三维培养模型在80 mmol/L KCl刺激下可检测到的最大收缩力为(8.4±0.6)mg显著大于HSP27干扰组 [(3.8±0.2)mg,P<0.05]。      结论      HSP27低表达状态可导致BSMCs肌动蛋白结构变化并导致其收缩功能受损。

Abstract:

Objective      To investigate the effect of low expression of heat shock protein 27 (HSP27) on actin skeleton structure and contraction function of human bladder smooth muscle cells (BSMCs) in vitro.       Methods      To inhibit the expression of HSP27, 3 pairs of siRNA were designed, inserted into lentiviral vector pGreenPuro and transferred into human BSMCs. Phalloidin staining and F-actin/G-actin ratio detection were used to observe the actin skeleton structure and polymerization status in the cultured BSMCs. The bladder smooth muscle fibers were reconstructed by culturing BSMCs with collagens to evaluate the contractile force.       Results      Lentiviral vector pGreenPuro-sh-HSP27 was successfully constructed and transferred into the cultured BSMCs effectively. Low-expression of HSP27 caused significant disruption of actin skeleton structure and decrease of F-actin/G-actin ratio (0.116±0.001) in BSMCs compared with the normal control (8.717±0.500, P<0.05). The contractile force
(3.8±0.2 mg) of the reconstructed fibers was significantly attenuated by low-expression of HSP27 compared to the normal control (8.4±0.6 mg, P<0.05) in the stimulation of 80 mmol/L KCl.       Conclusion      Low expression of HSP27 can impair contractile force of human BSMCs through altering the actin skeleton structure.

参考文献/References:

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更新日期/Last Update: 2015-03-19