Effect Of Ibuprofen On Mice Ovarian Function

Ibuprofen is one of the most commonly used non-steroidal anti-inflammatory drugs,which has the functions as analgesics,anti-inflammatory and antipyretics.Previous studies have demonstrated that young men,who had taken Ibuprofen for a long time,could lead to disorders of sex hormone secretion,resulting in a decrease in sperm motility and other negative consequences.However,there is little information about the effects of Ibuprofen on female ovarian functions.Thus,the objectives of this study to use mice as an animal model to determine the effects of Ibuprofen on ovarian functions.The experiments were divided into two parts:Part ?: The manuscript explored whether Ibuprofen could cause damages to ovarian functions in the mice in vivo.To answer this question,fifty 6-week-old ICR female mice with similar body weight were randomly divided into 5 groups with 10 mice in each group.Mice in each group were daily injected Ibuprofen with a different concentration of DMSO,0 mg/kg,10 mg/kg,20 mg/kg and 40 mg/kg for 7days.No-injection control group and DMSO experimental control group were set.On day 8,bodyweight and ovarian weight of each mouse were measured,respectively.Ovaries were collected and paraffin sectioning,HE staining andimmunohistochemical staining were performed with these collected ovaries.The number of healthy and atretic follicles in ovaries were carefully observed and analyzed.Moreover,RT-qPCR was conducted to examine the expression level of Gdf9,Scp3 and Foxl2 in the ovaries and the level of estrogen secretion in serum collected from these mice were measured.The results showed that the number of healthy follicles in the Ibuprofen groups decreased with the number of atretic follicles increased.Moreover,the serum estradiol and progesterone concentrations decreased and the expression level of Gdf9,Scp3 and Foxl2 decreased and the reproductive capacity of female mice of the 40 mg/kg group decreased significantly.The above results showed that ibuprofen inhibited the ovarian function of mice in vivo,and the inhibition was positively correlated with the injection concentration of ibuprofen.Part ?: The manuscript explored the effect of ibuprofen on apoptosis of granulosa cells in vitro.In this part,different concentrations of ibuprofen(0 ?M,10?M,100 ?M and 200 ?M)were added to the cell culture medium to culture primary mouse ovarian granulosa cells for 24 hours in vitro.The apoptotic levels of granulosa cells were analyzed by cell morphology,Hoechst33342/PI fluorescence staining,RT-qPCR and Western blot.The results showed that the levels of nuclear breakdown,chromatin condensation,expression of apoptotic proteins Bax and Caspase-3increased significantly with ibuprofen treatment.The above results showed that ibuprofen promoted the apoptosis process of granulosa cells in vitro,and the level of apoptosis was positively correlated with the concentration of ibuprofen.In conclusion,this study shows that ibuprofen could promote the apoptosis process of follicular granulosa cells and impair ovarian function in ICR female mice.In addition,the degree of ovarian dysfunction was positively correlated with the concentration of ibuprofen treatment.Therefore,according to the above results,long-term use of ibuprofen is likely to cause damages to ovarian function in women,suggesting that high dosage of ibuprofen should be avoided in the long-term use of ibuprofen to avoid impaired reproductive capacity.