| Pelvic organ prolapse(POP) is defined as the descent of the pelvic organ into the vagina, which mainly results from dysfunction of the pelvic floor support. It seriously affects the physical and mental health of patients. Its definite mechanism is still not very clear. In recent years, many scholars have paid attention to the research on the mechanism of pelvic floor nerve injury and the problems to be solved urgently.Studies at home and abroad have confirmed that POP and stress urinary incontinence(SUI)is closely related to the nerve injury of the pelvic floor and the local microenvironment of nerve plays a critical role in the regeneration process after pelvic floor nerve injury. It is bound to affect the regeneration and repair of pelvic nerve for the microenvironment changing. Neural cell adhesion molecule(NCAM) is one of the important component of the neural microenvironment and it is involved in the process of nerve growth and development, recognition, aggregation repair and regeneration,and so on. NCAM-180 isoform is mainly distributed in the mature nerve cells,which can inhibit the growth of signal response to inhibit nerve regeneration, increase the adhesion between cells, maintain the stability between nerve and the surrounding tissues. Its abnormal expression will lead to permanent damage of nerve. Neurofilament protein(NF) is the main component of the nerve cytoskeleton and the structural protein of the neuron specific. It has the function of "scaffolding", and it is able to maintain the normal form and function of neuron, to promote the growth of the axon, and determine and extend the axonal diameter.Its distribution and structure change reflect the structure and function of neurons and affect the connection rate of the nerve,the diameter, density and total number of the regenerated nerve fibers.ObjectiveBy studying of the expression of NF,NCAM-180 isoform in the anterior vaginal wall, we preliminarily exploreres their function and mechanism in POP, expecting to provide the path and basis for prevention, treatment and research of POP.Materials and Methods1. Materials:We selected 58 cases of POP-Q grade III and above patients without SUI from June 2013 to February 2015 in the department of gynaecology of our hospital as the POP group, including 29 cases of premenopausal and 29 cases of postmenopausal women,and operated by total vaginal hysterectomy & anterior and posterior wall repair or pelvic floor reconstruction;meanwhile 39 cases of non hormone related diseases, cervical cancer precancerous lesions or early cancer patients without pelvic floor dysfunction in the corresponding period as control group,including 17 cases of premenopausal and 22 cases of postmenopausal women,and operated by Total abdominal or vaginal hysterectomy. There was no statistical significance in the comparison of general data of patients of the two corresponding groups(P>0.05). All patients in the past 3 months without systemic application of hormone drugs and they were no hormone related diseases, nervous system, chronic respiratory disease, metabolic disease and connective tissue disease, and so on menopausal years about postmenopausal women were more than 2 years. Approved by the ethic discussion group and signed the agreement with the consent of the patients and their family members. We took the anterior vaginal wall tissue of the vaginal vault about 1.0cm *1.0cm size,nearly 12 o’clock direction.2. MethodsHE staining was applied to observe the tissue morphological characteristic of the anterior vaginal wall tissue; immunohistochemical staining(SP method) was used to test the expression of NF, NCAM-180,and western blot(WB)was applied to observe the expressions of NCAM-180 from molecular level.3. Statistical analysisUse SPSS17.0 statistical software for data processing. The c±sform was used to express the count data. Two independent samples t test was used to compare the groups. Pearson or Spearman correlation analysis was used to analyze the dependency of NF, NCAM-180 and parameters, and the correlation between the two. Alpha = 0.05.Results1 The histopathological characteristic of HE staining in each group and the anterior vaginal wall tissue:HE staining observed under microscope(×400) in control group: normal microvascular morphology was normal, complete structure, thick nerve fiber bundles and thick smooth muscle bundles, and arranged in neat rows, collagen fibers arranged neatly. POP group: structure of the lamina propria of loose and disordered, microvascular reduced and the tube wall was thinner, nerve fiber bundle was thinning and atrophy, connective tissue and smooth muscle bundle was fine and fracture.2 The immunohistochemical staining resultsIn the anterior vaginal wall tissue, the positive expression of NF, NCAM-180 is yellow or yellow brown; NF, NCAM-180 gray value was tested by Leica owin system,the intensity of immunohistochemical staining was proportional to the gray value. The positive substance was expressed higher, dyeing deeper, and gray value is greater, otherwise smaller.1) The gray value that NF expressed in the anterior vaginal wall tissue of he premenopausal pop group and control group, postmenopausal pop and control group patients respectively is(107.94±7.58);(167.25±8.11);(89.83±3.16);(142.9±4.90), The corresponding POP group were significantly lower than the control group,both had statistical difference(t=-16.90,P = 0.001;t=-28.81, P =0.001).2) The gray value that NCAM-180 expressed in the anterior vaginal wall tissue of he premenopausal pop group and control group, postmenopausal pop and control group patients respectively is(186.08.±14.71),(86.78±17.29),(141.00±12.36),(59.91±10.13), The corresponding POP group were significantly higher than the control group,both had statistical difference( t=13.83,P =0.001;t=16.05,P =0.001)3)The correlation analysis between the expression of NF、NCAM-180 and the parameters.The expression of NF was negatively correlated with age and menopausal status(r=-0.353, p=0.025; r=-0.429, p=0.006);The expression of NCAM-180 was negatively correlated with age and menopausal status(r=-0.365, p=0.021; r=-0.403, p=0.010).4) The correlation analysis that between NCAM-180 and NF.The expression of NCAM-180 was negatively correlated with the expression of NF(r=-0.718,p=0.001).3 Western blot resultsUsing GAPDH as internal control,the average integral optical density of NCAM-180 and GAPDH was tested by Bandscan5.0software. NCAM-180/GAPDH ratio was used to express the relative levels. Compared with the control group. The expression level of NCAM-180 in premenopausal POP group increased 38.71%(P<0.01), and in postmenopausal POP group increased 91.67%(P<0.01).Conclusions1 The high expression of NCAM-180 and low expression of NF in the vaginal wall of patients in POP groups, suggest that nerve injury or repair might be involved in the occurrence of POP.2 The expression of NF and NCAM-180 was negatively correlated with age and menopausal status,for elderly patients with POP, the effect of low estrogen levels and nerve injury on POP, need to be further studied. |
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