Role of Tibolone and Cimicifuga Racemosa on Urinary BladderAlterations in Surgically Ovariectomized Adult Female Rats

Authors : Mona H. Mohammed Ali, Wael Amin M. S. Nasr El-Din and Ghada A. Abdel-Hamid

DOI :

Volume : 38

Issue : 2

Year : 2015

Page No : 1-24

Introduction: Estrogen hormone is essential for the physiological maintenance of the female genitourinary tracts. Postmenopausal women are subject to several urologic dysfunctions. Due to the long-term health risks associated with hormone replacement therapy, many menopausal women are actively seeking alternative treatments. Cimicifuga racemosa has an estrogenic effect affording beneficial effects in menopause. Aim of work: To clarify the effect of estrogen hormone deficiency on the urinary bladder structure as well as to study the role of administration of tibolone and cimicifuga racemosa on these effects in a rat model of experimentally induced menopause. Materials and methods: A total of 50 adult female rats were used in this study. The animals were randomly divided equally into 6 groups of 10 each. Group 1 (Control group). Group II (Ovariectomized group):where rats were bilaterally ovariectomized. Group III (Ovariectomized plus tibolone-treated group): where rats were bilaterally ovariectomized and treated with tibolone. Group IV (Ovariectomized plus cimicifuga racemosa-treated group): where rats were bilaterally ovariectomized and treated with cimicifuga racemosa. Group V (Cimicifuga racemosa-treated group): where rats were treated with cimicifuga racemosa. At the end of the experimental period, all rats were anaesthetized and sacrificed. The bladder was removed and processed to be stained with Haematoxylin & Eosin and Malloryʼs trichrome stains. These urinary bladder sections were further immunohistochemically analyzed using estrogen receptors alpha and beta (ERα &ERβ) in the bladder's musclosa, caspase 3 as an immunohistochemical marker of apoptotic activity in the bladders urothelium and Proliferating Cell Nuclear Antigen (PCNA) for the assessment of proliferative activity of the bladder`s urothelium. The bladder wall and urothelium thickness, the mean area percentages of collagen fiber (CF) and smooth muscle (SM) within the musculosa were measured and the CF/SM ratio was calculated. In addition, the urothelial apoptotic and proliferative indices were measured. All these measures were statistically analyzed. Results: Urinary bladder sections of the ovariectomized rats revealed focal reduction in the epithelial lining and the total thickness of the bladder wall. Some epithelial cells had vacuolated cytoplasm with peripherally placed nuclei. Many cells had deeply stained nuclei. Areas of complete denudation of the mucosa with desquamated epithelial cells within the lumen were evident. The musculosa was distorted in arrangement and showed wide separation of the muscle bundles. Abundant amount of collagen fibers were observed among muscle bundles. A massive increase in positive cytoplasmic immune-expression of caspase 3 proteins in cells was observed in the superficial, intermediate and basal layers of the urothelium accompanied with decreased nuclear PCNA immunostained cells in the basal and intermediate layers of the urothelium. A massive increase in the immunoexpression of positive ER-α in the muscle layer was found while the immunoexpression of positive ER-β cells was decreased in the muscle layer. Administration of both tibolone and cimicifuga racemosa resulted in improvement of these alterations with better effect of cimicifuga racemosa compared to tibolone. Conclusion: Estrogen deficiency has deleterious effects on the urinary bladder structure. Tibolone administration improved the alteration of the bladder structure to some degree, whereas supplementary estrogen in the form of cimicifuga racemosa could improve bladder histopathological changes better than tibolone. Therefore, cimicifuga racemosa may be considered a consistent and safe option to counteract urinary bladder alterations in menopausal practice.