CAJ | 학술논문

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)、1.001±0.029(10％组,P＜0.0001)、0.821 ±0.032(15％组,P＜0.0001)、0.621 ±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最活跃.
목적 탐색불동견장력대방광평활기세포형태화증식활성적영향. 방법 방광평활기세포복합규효재료후대재료시가5％、10％、15％、20％급25％등불동대소적견장력,공취초현미경하관찰세포형태변화.CCK-8매표법검측세포증식능력,류식세포의분석세포주기병계산세포증식지수. 결과 재견장력작용하방광평활기세포정현“수축형”형태학개변,유무서적성형서전상태향상대규칙적방추상세포형태천이변화.CCK-8소측대조조흡광치도A위0.471 ±0.027,각견장조분별증지1.320±0.094(5％조,P＜0.0001)、1.001±0.029(10％조,P＜0.0001)、0.821 ±0.032(15％조,P＜0.0001)、0.621 ±0.032(20％조,P=0.0004)급0.591±0.056(25％조,P=0.0268).여대조조(29.35±0.55)％상비,세포증식지수증가도(55.55±1.05)％(5％조,P＜0.0001),(47.70±0.20)％(10％조,P＜0.0001),(35.40 ±2.10)％(15％조,P＜0.0001),여20％조적(34.85±0.55)％(P =0.1372)화25％조적(30.35 ±0.45)％(P =0.5234)비교차이균무통계학의의. 결론 괄당강도견장력가이조절방광평활기세포형태병촉진세포증식활성,5％형변솔적견장력하세포증식최활약.
Objective To investigate the effects of amplitude dependent morphological and proliferative changes in human bladder smooth muscle cells (hBSMCs) undergoing physiological stretch in vitro.Methods The hBSMCs were cuhured 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,cyclically 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 morphological 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 (control) 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 index 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 proliferative potential could be seen at 5％ of stretch.