Avaliação do potencial da cimicifuga racemosa (l.) Nutt. em reduzir os efeitos adversos causados pela dexametasona e doxorrubicina em folículos ovarianos de camundongas cultivados in vitro

Abstract

Considering Cimicifuga racemosa (L.) Nutt. (CIMI) as an alternative and natural drug in the treatment of menopause, as well as the importance of the study of drug redirection, this study aims to investigate the action of CIMI as a potential factor capable of reducing the deleterious effects of dexameasone (DEXA) and doxorubicin (DOXO). The experimental studies were approved by CEUA of the Federal University of Ceará Campus Sobral under protocol N◦ 05/18. For that, Swiss female mice (n = 47) with regular estrous cycle were used. In the laboratory, the ovaries were collected and cultured individually in a 24-well plate at 37.5 ° C, in 5% CO2, for 6 days. For experiment 1 of this dissertation, the following groups were evaluated - (i) DMEM +; (ii) CIMI [5 ng/ml]; (iii) DOXO [0.3 μg/ml]; (iv) CIMI [5 ng/ml] + DOXO [0.3 μg/mL]. For experiment 2, the following treatments were investigated: (i) DMEM +; (ii) CIMI [5 ng/ml]; (iii) DEXA [4 ng/ml]; (iv) CIMI [5 ng/ml] + DEXA [4 ng/ml]. After culture, the ovaries were used for the analysis of survival, activation, growth and follicular viability, ECM, apoptosis rate and stroma density. The results of activation, survival and growth were compared using Fisher's exact test, and the differences were considered significant when p <0.05. In experiment 1, the results showed that the presence of DOXO in culture medium, significantly reduces the percentage of morphologically normal follicles. On the other hand, the presence of CIMI [5 ng/ml] alone can maintain the percentage of morphologically normal follicles like the control group (DMEM+) and when added to the medium containing DOXO [0.3 μg/ml]. In experiment 2, the presence of DEXA [4 ng/ml] reduces the percentage of morphologically normal follicles when compared to the control group, while the presence of CIMI [5 ng/ml] can keep the percentage similar to the control group and still attenuate the deleterious effects caused by the presence of DEXA in culture medium. Regarding follicular activation and development, the ovaries when cultured in the presence of DEXA or DOXO reduced the percentage of primordial follicles compared to the control group, while the presence of CIMI [5 ng/ml] alone, kept the percentage of primordial follicles equal to group control. The presence of DEXA or DOXO negatively affected the stroma density, while the presence of CIMI [5 ng/ml] in the culture medium protected against the damage caused by these drugs. In addition, the presence of DEXA or DOXO increased the rate of apoptosis, while CIMI [5 ng/ml] in the culture medium reduced apoptosis. In view of the above, we can conclude that the CIMI extract [5 ng/ml] was able to contain the deleterious effects illustrated by DEXA and DOX at the end of 6 days of culture.