The Expressions And Clinical Significances Of Collagen â…¢ And MMP-3 In Pelvic Floor Dysfunction Women's Carainal Ligament Tissue

Objective: Pelvic floor dysfunction (PFD) is a kind of comom gynecologic diseases,and it mainly occurs in middle and old aged women.For the psat few years,the disease rate of PFD has raising up along with the aging society.The disease has caused a world of harm to many women's body and mind.With regard to many medical workers, we should identify the pathogenesis of PFD. It will make us diagnose and cure the disease better. Present work aimed to detect the expression levels of Collagen III and MMP-3 which can decompose the Collagen III in the patients' cardinal ligment of uterus,and then analyze the relation of collagen decompositive metabolism ,and the level of expression of MMP-3 in the development of PFD.Methods:1 Object of research:the examples were selectde from March 2009 to December 2010 in Hebei Province People's Hospital of department of gynecology.The diagnostic standard is accord with POP-Q.All the PFD patients were III and IV periods of POP.26 patients of PFD were conducted as the experimental group,and among thpat6 patients.The stada16 patients were premenopausal patients and 10 patients were postmenopausal patients.24 patients without PFD who were operated the hysterectomy because of genecologic benign lesion were conducted as the control group.Among the 24 pationts,14 patients of them were premenopausal and 10 patients of them were postmenopausal.All of the research objects were without the abnormal elastic fibre disease,such as scarify skin-disease,emphysema,and moyamoya disease. They were without the genital duct and urinary infections. All of the premenopausal patients' endometrium were in the proliferative phase. All of the research objetcts had not taken the hormonal medicine in the near 3 months and pathologic results after operations were without correlative estrogen diseases such as endometriosis.2 Methods: The cardinal ligment tissues of uterus were taken in the operation(0.5×0.5×0.5cm30.5×0.5×0.5cm3),fix in the formaldehyde,embedded in the paraffin routinely,and some of the tissue slice were conduted with HE staining,so that,we can identify the histologic structures.All of the research objects had signed the informed consents before the operation.We applied immunohistochemisty method to detect the expression levels of the MMP-3 and Collagen III in the cardinal ligment tissues of uterus in the 4 groups.All of them were 50 patients,16 premenopausal patients with PFD, 10 premenopausal patients without PFD, 14 postmenopausal patients with PFD,and 10 postmenopausal patients without PFD.3 Reagents:The main immunohistochemical reagents are rabbit anti-human III collagen monoclonal antibody, rabbit anti-human MMP-3 polyclonal antibody, SP kit and DAB liquid chromogenic enzyme substrate kit, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride , and hydrogen peroxide.4 Image Analysis of immunohistochemistry:①.Select the field of vision of 400×,select randomly the 5 fields of vision from every immunohistochemistry slice,and take 5 photos from every field of vision.②To use the Motic Med 6.0 computer to analyze positive aeras average grayscale of MMP-3 and CollagenIII immunohistostaining of the cardinal ligament.And then,calculate the average value.5 Analyze the data with SPSS13.0.Results:1 The main pathogenetic influencing factors of PFDThe main pathogenetic influencing factors of PFD are the age,the number of times of childbirth, BMI,and the age of menopause.There were no significant differences with the main pathogenetic influencing factors above through the T-analysis(P> 0.05),that proved that there were comparability between the two groups. 2 Light microscope showed:(HE staining) : Experimental group of women's the uterus cardinal ligament tissues showed the collagen density decreased,alinement was discrete and fine crushing.There were blood vessel and smooth musle issostructures.Cells arranged loosely.Some of the nucleus were pycnosis and spiltting. Control group of women's the uterus cardinal ligament tissues showed the collagen fibres was pink,staining well-distributed,and content abundance.They arranged bunchly and braidingly.The cells arranged regulately and condensingly. The nucleus were blue-stain,and arround the collagen fibre.There were no pycnosis and splitting phenomenons.3 The results of immunohistochemistry3.1 The comparison of MMP-3 expressionMMP-3 mainly locates in the hyalomitome,and the positive reactant is buffy colar.Observe every slice in the high power mirror,and use the computer system to measure the average grayscale value.The MMP-3 average grayscale value of the premenopausal group with PFD was 89.76±7.86,and the control group of premenopause was 73.68±7.02. Pair T-test analysis of statistical showed that: the PFD group and control group, P = 0.01 (P <0.05), there are significant differences.The MMP-3 average grayscale value of the postmenopausal group with PFD was 106.23±4.28,and the control group of postmenopause was 91.29±7.16. Pair T-test analysis of statistical showed that: the PFD group and control group, P = 0.01 (P <0.05), there are significant differences.3.2 The comparison of CollagenIII expressionCollagenIII mainly locates in the nucleus and hyalomitome,and the positive reactant is buffy colar.Observe every slice in the high power mirror,and use the computer system to measure the average grayscale value.The CollagenIII average grayscale value of the premenopausal group with PFD was 89.59±9.89,and the control group of premenopause was 102.06±7.76. Pair T-test analysis of statistical showed that: the PFD group and control group, P = 0.003 (P <0.05), there are significant differences. The CollagenIII average grayscale value of the postmenopausal group with PFD was 79.86±7.98,and the control group of postmenopause was 90.24±6.67. Pair T-test analysis of statistical showed that: the PFD group and control group, P = 0.01 (P <0.05), there are significant differences. 3.3 The correlation analyses of the positive expressions of CollagenIII and MMP-3 in the premenopausal PFD groups.We applied Pearson to analyze the the associativity of expressions of CollagenIII and MMP-3 of premenopausal PFD group.The results indicated that the rates positive expressions of CollagenIII and MMP-3 showed significant negative correlation in the premenopausal PFD group. r=-0.719 P=0,002 (P <0.05)We applied Pearson to analyze the the associativity of expressions of CollagenIII and MMP-3 of postmenopausal PFD group.The results indicated that the rates positive expressions of CollagenIII and MMP-3 showed significant negative correlation in the premenopausal PFD group. r=-0.994 P=0,000 (P <0.05)Conclusions:1 The CollagenIII positive expression rate of PFD group was more decreasing compared with the control group no matter premenopause or postmenopausal. It caused the connective tissue which contains collagen mainly in the cardinal ligament of uterus decreasing.So the supportive strength for the uterus has been weakened,and can't keep the uterus at the normal dissection level.Through this mechanism,it participates the development of PFD.2 In the experiment , the MMP-3 positive expression rate of PFD group was more increasing compared with the control group no matter premenopause or postmenopausal,that possibly caused the collagen decomposition increasing.Accordingly,it can make the supportive tissue of pelvic floor damaged.It is the cause of the generation and development of PFD.3 The average grayscale value of CollagenIII and MMP-3 showed significant negative correlation in the premenopausal and postmenopausal PFD groups.It demonstrates that MMPs can decompose collagen.It makes the supportive structure of pelvic floor decrease,and then causes the ligament and fascia of pelvic floor loose,which makes PFD develop.