Effects of Huoxue Xiaoyi Decoction on Apoptosis of Granulosa Cells in Endometriosis Rats

Abstract Background: Endometriosis(EM) is a common disease that occurs in reproductive age. 50% endometriosis patients is suffering from infertility. Follicular development is the main cause of endometriosis-associated infertility. Here the study based on apoptosis of granulosa cells during follicular development will explore the effect and the possible mechanism of Huoxue Xiaoyi Decoction (HXXYD) on apoptosis of ovarian granulosa cells in endometriosis model rats. Methods: Thirty 8-week-old female SD rats were divided into four groups: blank group, sham-operation group, model group and HXXYD group. Blank group, sham-operation group and model group were given double-distilled water, while HXXYD group were given HXXYD for 15 days. After intragastric administration, blood samples from abdominal aorta of rats were collected to detect oxidative and antioxidative indexes including ROS, T-SOD, CAT. The morphology of follicles were observed by H&E staining and every stage of follicles were calculated. The location of granulosa cells and apoptosis related factors including Bax, Bcl-2, caspase-3 were stained by immunohistochemistry staining. The apoptosis of granulosa cells were stained by TUNEL staining and the rate of apoptosis were calculated. Apoptosis related proteins including p-JNK, Bax, Bcl-2, caspase-3 were detected by Western blot. Results: The level of serum ROS decreased, and the levels of serum T-SOD and CAT increased in the HXXYD group. The number of secondary follicles increased in HXXYD group. The expression of Bax, caspase-3 in ovarian granulosa cells decreased and the expression of Bcl-2 increased in the HXXYD group with immunochemical staining. The apoptosis rate of ovarian granulosa cells in the HXXYD group decreased. The expression of p-JNK, Bax and caspase-3 protein decreased, the expression of Bcl-2 increased in the HXXYD group.Conclusions: These results indicate that HXXYD may improve the oxidative stress state, decrease the apoptosis of ovarian granulosa cells, and improve the development of follicles in endometriosis model rats through ROS-JNK signaling pathway.