Effect of bladder outlet obstruction on detrusor smooth muscle cell: an in vitro study.
BACKGROUND
Although relieving obstruction is generally curative on bladder outlet obstruction (BOO), bladder dysfunction persists in some patients. Repetitive stretch and relaxation applied to cultured bladder smooth muscle (SM) cells in vitro have been used to mimic increases in urodynamic load experienced by the detrusor muscle under conditions of BOO. We first clarified the relationship between phenotype transformation and biomechanical properties of detrusor smooth muscle cell (DSMC) subjected to the cyclic mechanical stretch.


METHODS
Cultured rat DSMC were grown on collagen-coated silicone membranes and subjected to continuous cycles of stretch-relaxation. All experiments were performed on cells between passages 2 and 4. Each cycle consists of 5 seconds of stretch and 5 seconds of relaxation. The computer controlled vacuum induced 10% (1), 20% (2), and 30% (3) maximum elongation of the plate membrane at different designed pressures. The deoxyribonucleic acid synthesis rate was assessed by performing tritiated thymidine incorporation assay. The expression of SM-alpha-actin and proliferation of DSMC were analyzed by immunofluorescent assay and flow cytometry. The image analysis and micropipet aspiration systems were used to investigate the single cell contraction and viscoelasticity. Using the 3-element standard linear solid model, the elastic modulus K(1), K(2), and viscoelastic coefficient mu were determined, which show the passive deformation ability of detrusor cells.


RESULTS
As the basic structural changes to mechanical stretch, DSMC undergo phenotypic modulation from their normal contractile phenotype to a "synthetic" phenotype: the DSMC become more proliferative and the actin less organized along the cell's long axis. The cell proliferation index of control and stretched group (10%, 20%, 30% elongation) are 0.24, 0.43, 0.58, and 0.65, respectively. The actin filaments in unstimulated cells were evident and orientated along the major axis of the cell. After mechanical stretch, the well-spread filaments changed their orientation. The function, such as contraction, and viscoelasticity of a single DSMC subjected to stretch both decreased significantly compared with control. The maximum contractile velocity and maximum cell length shortening rate of group 3 (30% elongation) showed significant decreases compared with unstretched control (P < 0.01). K(1) and K(2) were decreased with the increase of mechanical overload. However, there was no statistic difference between groups 2 and 3.


CONCLUSIONS
Functional abnormalitie of BOO have the structural basis: phenotype transformation (i.e., remodeling) of the detrusor cells. Cyclic stretch and relaxation applied to DSMC in vitro can be used to model increases in urodynamic load experienced by the bladder detrusor muscle under conditions of BOO. Phenotype transformation is the structural basis of functional changes of DSMC subjected to periodic overload mechanical stretch.
Yu
Gong
G
Urologic Center, Southwest Hospital, Third Military Medical University, Chongqing, People's Republic of China. gongyu2009@sina.com
Bo
Song
S
Xiyu
Jin
J
Enqing
Xiong
X
D000199:Actins / Q000235:genetics; D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division; D002478:Cells, Cultured; D004261:DNA Replication / Q000502:physiology; D004195:Disease Models, Animal; D004548:Elasticity; D008297:Male; D009119:Muscle Contraction / Q000502:physiology; D009126:Muscle Relaxation / Q000502:physiology; D009130:Muscle, Smooth / Q000166:cytology / Q000473:pathology / Q000503:physiopathology*; D051381:Rats; D017208:Rats, Wistar; D012828:Silicones; D001748:Urinary Bladder Neck Obstruction / Q000503:physiopathology*; D014783:Viscosity
D000199:Actins; D012828:Silicones