| Objective: Histological analysis of anterior vaginal wall in women with stress urinaryincontinence，and compare the smooth muscle contentand apoptosis of the vagina inwomen with and without stress urinary incontinence.Methods: Thirty biopsy specimens were obtained from the anterior vaginal wall ofwomen having a stress urinary incontinence (15patients with SUI combined POP) andthe same location in fifteen patients with no SUI from June to November2012inFuzhou general hospital. All subjects were divided into normal control group (controlgroup, n=15), urinary incontinence alone group (SUI group, n=15) and urinaryincontinence combined prolapse group (SUI+POP group, n=15), biopsy specimenswere obtained from the approximately the same location of anterior vaginal wall insurgery process. Routine H&E staining, Masson’s trichome staining, Aldehyde fuchsinstaining and immunohistochemical staining by CD31, S-100and actin antibodies wasused to qualitative analyze the microscopic structure and morphology properties inmore than one field. Morphometric analysis by ImagePro Plus6.0software wasperformed on histologic cross-sections of the anterior vaginal wall to determine thenumber and diameter of the sub-mucosal nerve fibers, the fractional area of vascular,nonvascular smooth muscle and connective tissue in the muscularis, and to analyze itscorrelation with clinical parameters of age, body mass index, the time of menopause,the combined degree of prolapse and VLPP. In addition, Apoptosis was assessed byterminal deoxynucelotidyl-transferase-mediated dUTP nick-end-labeling (TUNEL)method, mRNA and protein expression of anti-apoptotic and pro-apoptotic protein ofmitochondria apoptosis pathway (such as P53、BCl-2and Bax) in vaginal tissues fromsubjects and controls were determined by real-time PCR and Western blot，to analyzeits correlation with clinical parameters of age, body mass index, the time ofmenopause, the combined degree of prolapse and VLPP. Results:(1) The result of various staining showed that: morphometric characteristicswere significantly altered in women with stress urinary incontinence. Compared withnormal control group, patients with stress urinary incontinence vaginal muscularis isthinner, muscle bundle smaller, fewer in number, and poorly organised; collagen fibersscattered, irregular distribution; elastic fiber distribution less, and there are differentdegrees of fault; small number of peripheral nerve fibers, and smaller diameter.(2) Quantitative analysis results showed that:①The mean numbers of the nerve fibersof muscularis smooth muscle of women in control group, SUI group and SUI+POPgroup was respectively (3.1±1.4)，(2.2±1.3) and (1.8±1.0). Number of thesubmucosal nerve was significantly decreased in SUI group and SUI+POP groupcompared to women in control group(P＜0.05), but there was no statisticallysignificant difference between SUI group and SUI+POP group(P＞0.05).②Themean diameter of the nerve fibers of muscularis smooth muscle of women in controlgroup, SUI group and SUI+POP group was respectively (42.1±7.2),(33.2±6.8) and(31.8±4.7)μm. Diameter of the submucosal nerve was significantly decreased inSUI group and SUI+POP group compared to women in control group(P＜0.05), butthere was no statistically significant difference between SUI group and SUI+POPgroup(P＞0.05).③The mean fractional area of blood vessels of muscularis smoothmuscle of women in control group, SUI group and SUI+POP group was respectively(2.2±1.0)，(2.5±1.1) and (2.4±1.2)%. There was no statistically significantdifference among three groups(P＞0.05);④The mean fractional area of nonvascularsmooth muscle of women in control group, SUI group and SUI+POP group wasrespectively (56.9±9.4)，(47.7±6.9) and (44.3±6.0)%. Fractional area ofnonvascular smooth muscle was significantly decreased in SUI group and SUI+POPgroup compared to women in control group (P＜0.05), but there was no statisticallysignificant difference between SUI group and SUI+POP group(P＞0.05);⑤Themean fractional area of connective tissue of women in control group, SUI group andSUI+POP group was respectively (40.3±8.4)，(50.7±7.3) and (54.9±6.6)%.Fractional area of connective tissue was significantly increased in SUI group and SUI+POP group compared to women in control group(P＜0.05), but there was nostatistically significant difference between SUI group and SUI+POP group(P＞0.05);⑥There was statistically significant correlation between some clinical parameters(such as age, parity, the numbers of the vaginal deliveries, the time of menopause,stage of prolapse and VLPP) and the numbers or diameter of the nerve fibers, and themean fractional area of blood vessels of muscularis smooth muscle showed correlationwith some clinical parameters (such as age, parity, the numbers of the vaginaldeliveries, the time of menopause). In addition, there was statistically significantcorrelation between all clinical parameters in our study and the mean fractional area ofnonvascular smooth muscle or connective tissue in the anterior vaginal wall frompatients.(3) TUNEL staining and apoptosis-related protein expression analysis showed that:①The the apoptotic index of nonvascular smooth muscle of women in control group,SUI group and SUI+POP group was respectively (9.8±2.2),(18.1±4.7) and (20.2±5.4). The apoptotic index was significantly increased in SUI group and SUI+POPgroup compared to women in control group(P＜0.05), but there was no statisticallysignificant difference between SUI group and SUI+POP group(P＞0.05);②mRNAexpression of P53, Bcl-2and the ratios of Bcl-2/Bax was upregulated, but mRNAexpression of Bax was lower in control group versus cases group；③Proteinexpression of Bcl-2and the ratios of Bcl-2/Bax was higher, but protein expression ofP53and Bax was lower in control group versus cases group，and there was nostatistically significant difference between SUI group and SUI+POP group(P＞0.05)；④There was statistically significant correlation among protein expression of P53,Bcl-2, Bax, the ratios of Bcl-2/Bax and the apoptotic index. There was statisticallysignificant correlation between BMI and the protein expression of P53, and the proteinexpression of Bcl-2showed correlation with age, the time of menopause and stage ofprolapse; the protein expression of Bax showed correlation with age, parity, thenumbers of the vaginal deliveries, stage of prolapse and VLPP. In addition, there wasstatistically significant correlation between all clinical parameters in our study and theratios of Bcl-2/Bax and the apoptotic index. Conclusion:(1) The nerve and muscle degenerative changes of the vaginal wall in women withSUI may be a potential pathogeny of SUI.(2) Nonvascular smooth muscle apoptosis of vaginal anterior wall may contribute todevelopment of pelvic floor disorders.(3) The mitochondrial apoptosis pathway maybe was involved in the regulation ofapoptosis process... |
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