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目的研究4种不同生物力学应力施加装置对体外培养的膀胱平滑肌细胞生长的影响,设计可行的应力施加装置。方法本研究设计了4种生物力学装置(圆形压力罐,玻璃水柱管,压力培养瓶,张力罐),将体外培养的膀胱平滑肌细胞分为实验组和对照组,实验组施予40cmH2O(1cmH2O=0.0981kPa)持续性的静水压力(或7.7N牵张力),对照组不施予任何压力,24h后用胆囊收缩素8肽(CCK8)法检测细胞增殖的变化,免疫荧光检测膀胱平滑肌细胞骨架α-actin,RT-PCR检测增殖细胞核抗原(PCNA)表达的变化。结果每种装置加压组与对照组之间,以及4种装置加压组之间细胞CCK8吸光度差异无统计学意义(P>0.05),细胞骨架α-actin表达差异无统计学意义(P>0.05),PCNA mRNA表达差异也无统计学意义(P>0.05)结论4种加压装置均对细胞无毒,40cmH2O(或7.7N牵张力)压力不能促进细胞增殖,加压装置在细胞培养和加压方式等方面具有多种优点。
Objective To study the effects of four different biomechanical stress devices on the growth of bladder smooth muscle cells in vitro and to design a feasible stress applying device. Methods Four kinds of biomechanical devices (circular pressure tank, glass water column, pressure flask and tension tank) were designed in this study. Bladder smooth muscle cells cultured in vitro were divided into experimental group and control group. The experimental group was given 40cmH2O (1cmH2O = 0.0981kPa). The rats in the control group were not subjected to any pressure for 24 hours. The changes of cell proliferation were detected by CCK8 assay after 24 hours, and the changes of the proliferation of bladder smooth muscle cells were detected by immunofluorescence α-actin, RT-PCR detection of proliferating cell nuclear antigen (PCNA) expression changes. Results There was no significant difference in the absorbance of CCK8 between the pressurization group and the control group and the compression group of the four devices (P> 0.05). There was no significant difference in the expression of α-actin in the cytoskeleton (P> 0.05). There was also no significant difference in the expression of PCNA mRNA (P> 0.05) .Conclusion The four kinds of pressurization devices are non-toxic to cells and the pressure of 40cmH2O (or 7.7N stretch) can not promote cell proliferation. There are many advantages such as pressure way.