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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

, PP. 500-504

Keywords: 热休克蛋白,肌动蛋白,逼尿肌细胞,膀胱收缩功能障碍

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Abstract:

目的探讨热休克蛋白27(heatshockprotein27,HSP27)低表达对体外培养人膀胱逼尿肌细胞(bladdersmoothmusclecells,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,Westernblot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、Westernblot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80mmol/LKCl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。

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