Expression And Significance Of Lysyl Oxidase And Collegan â…  In Cardinal Ligaments And Anterior Vaginal Wall Of Pelvic Organ Prolapse

ObjectiveTo detect the expression of lysyl oxidase,collagenⅠin cardinal ligament and anterior vaginal wall of pelvic organ prolapse,and discuss the role of them in the occurrence and development in pelvic organ prolapse.Method1.Tissue samples and groupsSixty-eight POP and thirty-eight NON-POP cardinal ligament and anterior vaginase tissue specimens were obtained from patients underwent surgical resection at Shengjing Hospital of China Medical University between 2007 May and 2008 Nov.All the samples were fixed by formalin and embedded in paraffin.2.ReagentsThe anti-human LOX rabbit polyclonal antibody were produced in Novus Biological Corporation,and anti-human Collagen TypeⅠrabbit polyclonal antibody were produced in Wuhan Boster Biotechnology Corporation.S-P kit,DAB agent kit and horseradish peroxidase labeled goat-anti-mouse IgG were bought from Beijing Zhongshan Golden Bridge Biotechnology Corporation.3.ImmunohistochemistryImmunohistochemical staining was performed by the streptavidin-peroxidase(S-P) method.The sections were deparaffinized,water processed,treated by pepsin to expose antigen at room temporate and incubated with primary antibody at 4.0℃overnight.The color was developed with diaminobenzidine and the sections were counterstained with hematoxylin.For the nonspecific reagent control,the primary antibodies were replaced by PBS. 4.Results assessmentUse immunohistochemical semiquantitative method:choose 5 high power field randomly(×400) and then score the staining intensity and staining area respectively. staining intensity:no staining record as 0 score,pallide-flavens as 1 score,yellow as 2 scores,buffy as 3 scores.Percentage of staining area:without staining area record as 0 score,≤20%as 1 score,20%～40%as 2 scores,40%～65%as 3 scores,≥65%as 4 scores.Subjugating the two scores,determining the results as weighting staining intensity,0～2 record as(-),2～4 as(+),4～6 as(++);6～12 as(+++).For the results assessment is according to the staining reaction of most of the cells and the mesenchyma,as to reduce subjective one-sidedness,every section was examined by 3 pathologist respectively,and then verified their results,found the common ones.5.Statistical analysis.The statistical package SPSS13.0 was used for all analyses.The chi-square test was used to determine the correlation between LOX or collagenⅠexpression,the relationship between LOX and collagenⅠwas analized by pearson test.Values of P＜0.05 were considered statistically significant.Results1.The expression of LOX in cardinal ligament and anterior vaginal wall of POP and control group:The LOX positive expression rate in cardinal ligament and anterior vaginal wall of pre-menopausal and postmenopausal POP group is 29.6%,22.2%,46.3%,41.5%, respectively.The LOX positive expression rate in cardinal ligament and anterior vaginal wall of pre-menopausal and postmenopausal control group is 66.7%,73.9%,73.9%,69.6%,respectively.The expression of difference of LOX in premenopausal POP group and the control group of the cardinal ligament and anterior vaginal wall is significant(x~2=4.659,P＜0.05;x~2=4.200,P＜0.05).The expression of difference LOX in postmenopausal POP group and the control group of cardinal ligament and anterior vaginal wall is significant(x~2=4.047,P＜0.05;x~2=9.329,P＜0.05). 2.The expression of collagenⅠin cardinal ligament and anterior vaginal wall of POP and control group:The collagenⅠpositive expression rate in cardinal ligament and anterior vaginal wall of pre-menopausal and postmenopausal POP group is 33.3%,25.9%,39.0%,34.1%,respectively.The collagenⅠpositive expression rate in cardinal ligament and anterior vaginal wall of pre-menopausal and postmenopausal control group is 66.7%,65.2%,65.9%,73.9%,respectively.The expression of difference of collagenⅠin premenopausal POP group and the control group of the cardinal ligament and anterior vaginal wall is significant(x~2=4.325,P＜0.05;x~2=4.747,P＜0.05).The expression of difference collagenⅠin postmenopausal POP group and the control group of cardinal ligament and anterior vaginal wall is significant(x~2=4.047,P＜0.05;x~2=9.329,P＜0.05).3.The relationship between the expression of lysyl oxidase and collagenⅠin cardinal ligament and anterior vaginal wall of POP group:A positive correlation was observed between the expression of lysyl oxidase and collagenⅠin cardinal ligament and antetior vaginal wall of pre-menopausal pop group (rs=0.606,P＜0.01;rs=0.580,P＜0.01);And the same results was found in POP postmenopausal group(rs=0.341,P＜0.05;rs=0.371,P＜0.05).Conclusion1.The expression of lysyl oxidase in cardinal ligament and antetior vaginal wall of POP group in the pathogenesis of POP was decreased might play a role.2.The decreased expression of CollagenⅠin cardinal ligament and antetior vaginal wall of POP group may contribute to reducing the occurrence of the POP.3.The expression of LOX and collagenⅠin cardinal ligament and antetior vaginal wall of POP group in reducing the occurrence of the POP have obvious relevance.