Effect of penetrative needling of "Zhibian" (BL54) through "Shuidao" (ST28) on expression of internal and external apoptosis-related factors in ovarian of mice with poor ovarian response.

Abstract

Objectives: To observe the effect of penetrative needling of "Zhibian" (BL54) through "Shuidao" (ST28) on the expressions of internal and external apoptosis-related factors in mice with poor ovarian response (POR), so as to explore its mechanisms underlying improvement of POR.

Methods: Sixty-nine female C57BL/6N mice were randomly divided into blank, model and acupuncture groups (n=23 in each group). The POR model was established by gavage of tripterygium wilfordii polyglucoside suspension (50 mg·kg^-1). After successful modeling, the rats in the acupuncture group received penetrative needling of BL54 through ST28, with the needle retained for 20 min, once a day for 14 days. Thirty healthy male mice of the same strain aged 7-8 weeks were co-housed with female mice in a 1∶1 ratio. Vaginal exfoliative cell smears were used to observe the changes in the estrous cycle of female mice. The number of follicles were observed under light microscope, wet weight of ovaries was recorded and ovarian index was calculated. The pregnancy rate and number of embryos were observed. ELISA method was used to detect the low levels of anti Mullerian hormone (AMH), follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) contents in the serum of female mice. HE staining was used to observe the morphology of ovarian tissue. TUNEL staining was used to observe the apoptosis rate of ovarian cells. The mRNA and protein expression levels of cysteine-aspartic acid protease (Caspase)-3, Bcl-2-associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), fas cell surface death receptor (Fas), and fas-associated death domain (FADD) in ovarian tissue were detected by real-time quantitive PCR and Western blot, respectively.

Results: Compared with the blank group, the rate of estrous cycle disorder, the contents of FSH and LH in serum, the rate of ovarian apoptosis, the mRNA and protein expression levels of Caspase-3, Bax, Fas and FADD in ovarian tissues were significantly increased (P<0.01); whereas the pregnancy rate, the number of embryos, the number of eggs obtained, ovarian wet weight, ovarian index, the contents of AMH and E2, the mRNA and protein expression levels of Bcl-2 were significantly decreased (P<0.01) in the model group. Following the intervention, the above-mentioned indexes were all reversed in the acupuncture group (P<0.01, P<0.05).

Conclusions: Penetrative needling of BL54 through ST28 can increase the ovarian response in POR mice, which may be associated with its function in regulating the expression of internal and external apoptosis-related factors in ovarian tissue.