Effects of moxibustion on delaying ovarian aging in mice by activating mitochondrial unfolded protein response and enhancing granulosa cell mitochondrial function

ObjectiveTo investigate the effect of moxibustion on delaying ovarian aging in naturally aging mice and to explore the underlying mechanisms.MethodsFifteen 2-month-old female SPF ICR mice with regular estrous cycles were included in a 2-month-old group. Thirty 10-month-old female SPF ICR mice were randomly divided into a 10-month-old group and a moxibustion group， with 15 mice in each group. The moxibustion group received alternating moxibustion on bilateral “Shenshu”（BL23） and “Zhongwan” （CV12）， “Guanyuan” （CV4） for 10 min every other day， for 21 consecutive days. The vaginal smears were used to observe the estrous cycle irregularities of mice. The serum contents of estradiol （E2）， follicle-stimulating hormone （FSH）， and anti-Müllerian hormone （AMH） were measured by ELISA. The ovarian morphology was observed by hematoxylin-eosin staining. The contents of adenosine triphosphate （ATP） in granulosa cells were measured by luminescence-based ATP assay. The levels of mitochondrial membrane potential （MMP） of granulosa cells were measured by JC-1 staining. The real-time fluorescence quantitative PCR， Western blot， and immunohistochemistry were used to assess the mRNA and protein expression levels of mitochondrial unfolded protein response （UPRmt）-related signaling pathway components， including activating transcription factor 5 （ATF5）， caseinolytic protease P （ClpP）， and Lon protease 1 （LONP1） in the ovarian tissues.ResultsCompared with the 2-month-old group， the 10-month-old group exhibited an increased rate of estrous cycle irregularity （PP2 and AMH contents （PPPPPP2 and AMH contents （PPPPPPConclusionMoxibustion can delay ovarian dysfunction related to natural aging in mice and modulate hormonal levels. Its mechanism may involve regulating the expressions of UPRmt-related signaling molecules such as ATF5， ClpP and LONP1， and enhancing mitochondrial function in granulosa cells.