摘要
目的探索不同牵张力对膀胱平滑肌细胞形态和增殖活性的影响。方法膀胱平滑肌细胞复合硅胶材料后对材料施加5％、10％、15％、20％及25％等不同大小的牵张力，共聚焦显微镜下观察细胞形态变化。CCK-8酶标法检测细胞增殖能力，流式细胞仪分析细胞周期并计算细胞增殖指数。结果在牵张力作用下膀胱平滑肌细胞呈现“收缩型”形态学改变，由无序的星形舒展状态向相对规则的纺锤状细胞形态迁移变化。CCK-8所测对照组吸光值度A为0．471±0．027，各牵张组分别增至1．320±0．094（5％组，P〈0．0001）、1．0014-0．029（10％组，P〈0．0001）、0．821±0．032（15％组，P〈0．0001）、0．6214-0．032（20％组，P=0．0004）及0．591±0．056（25％组，P=0．0268）。与对照组（29．35±0．55）％相比，细胞增殖指数增加到（55．55±1．05）％（5％组，P〈0．0001），（47．70±0．20）％（10％组，P〈0．0001），（35．40±2．10）％（15％组，P〈0．0001），与20％组的（34．85±0．55）％（P=0．1372）和25％组的（30．35±0．45）％（P=0．5234）比较差异均无统计学意义。结论适当强度牵张力可以调节膀胱平滑肌细胞形态并促进细胞增殖活性，5％形变率的牵张力下细胞增殖帚活阱．
Objective To investigate the effects of amplitude dependent morphological and prolife- rative changes in human bladder smooth muscle cells （hBSMCs） undergoing physiological stretch in vitro. Methods The hBSMCs were cultured on silicone membrane and stretched similarly to a bladder cycle at range of stretches and time. The elongation would increase up to 2.5% every 3 h and 5% （ or 10% , 15% , 20% and 25% depending on the experiment design） in the next 1 h, followed by a rapid decrease, cyclical- ly maintained for a total of 16 h. In subsequent 8 h （24 h cycles） the membrane was maintained in relaxed position. Immunofluorescence and confocal laser scanning microscope were employed to assess the morpho- logical changes. Cell counting kit-8 （CCK-8） and flow cytometry were used to assess the cell proliferation. Results The hBSMCs showed contractile phenotype after application of mechanical strain. Compared with control, the strains at 5% , 10% , 15% , 20% and 25% induced most of the cells to change from a more spread-out and stellate state with large cell surface contact areas to a typical spindle-like morphology. The orientation angle of BSMCs remarkably differed depending on the applied strain＇s magnitude. Absorbance value, which reflects the proliferation activity, analyzed by CCK-8 was improved from 0. 471 ±0. 027 （con- trol） to 1. 320 ± 0. 094 （5 % elongation group, P 〈 0. 0001 ） , 1. 001± 0. 029 （ 10% elongation group, P 〈 0. 0001 ） , 0. 821 ± 0. 032 （ 15% elongation group, P 〈 0. 0001 ） , 0. 621 ± 0. 032 （ 20% elongation group, P = 0. 0004） and 0. 591 ± 0. 056 （25 % elongation group, P = 0. 0268 ） , respectively. Cell proliferation in- dex increased from （29.35 ±0.55）% （control） to （55.55 ± 1.05）% （5% elongation group, P 〈 0.0001）, （47.70±0.20）% （10% elongation group, P〈0.0001）, （35.40＋2.10）% （15% elongation group, P 〈 0. 0001 ） , respectively. However, no significant difference was found in either 20% elongation group （34.85±0.55）% （P=0.1372） or 25% elongation group （30.35 ＋0.45）% （P=0.5234）. Conclusions Proliferative and morphological changes could be observed in hBSMCs in vitro. Maximal pro- liferative potential could be seen at 5% of stretch.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2012年第11期822-826,共5页
Chinese Journal of Urology
基金
国家自然科学基金（31170907）
关键词
膀胱
肌细胞
平滑肌
形态学
细胞增殖
牵张力
Urinary bladder
Myocytes, smooth muscle
Morphology
Cell proliferation
Stretch
