高齡引發卵丘細胞之粒線體異常

Hum Reprod. 2017 Oct 17:1-9. doi: 10.1093/humrep/dex309. [Epub ahead of print]

Age-related changes in the mitochondria of human mural granulosa cells.

Liu Y1, Han M2,3, Li X4,5, Wang H1, Ma M1, Zhang S1, Guo Y1, Wang S1, Wang Y1, Duan N6, Xu B1, Yin J6, Yao Y1.

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Abstract

STUDY QUESTION:

What changes in the mitochondria of human mural granulosa cells (mGCs) with maternal aging?

SUMMARY ANSWER:

The mitochondrial membrane potential (MMP) and the ability of oxidative phosphorylation (OXPHOS) of mGCs declines with reproductive aging, accompanied with more abnormal mitochondria.

WHAT IS KNOWN ALREADY:

Mitochondria play an important role in the dialogue between the mGCs and oocytes. However, the underlying mechanism of mitochondrial dysfunction in mGCs in aging is still poorly understood.

STUDY DESIGN SIZE, DURATION:

In total, 149 infertile women underwent IVF in the ART Centre of the Chinese PLA General Hospital, China from September 2016 to May 2017. Two age groups were investigated: the young group (<38 years old) and the old group (≥38 years old).

PARTICIPANTS/MATERIALS, SETTING, METHODS:

The mitochondrial ultrastructure of mGCs was observed by transmission electron microscopy, and real-time quantitative polymerase chain reaction was applied to quantify the mitochondrial DNA (mtDNA) copy number, 4977-bp deleted DNA and mRNA expression of mitochondrial ATP synthases ATP5A1 and ATP5I. MMP was detected by flow cytometry and fluorescence microscopy, respectively. Reactive oxygen species (ROS) was tested by flow cytometry. A luminometer was used to measure the ATP levels and western blot to analyse the OXPHOS complex.

MAIN RESULTS AND THE ROLE OF CHANCE:

In the young group, mitochondria were mostly round or oval, with a few intact parallel tubular-vesicular cristae and homogenous matrix density, while elongated mitochondria were mainly observed in the old group, which had numerous cristae and more high-density matrix particles. Abnormal mitochondria were more common in aging women (P = 0.012). mtDNA relative copy number was positively correlated with maternal age (r = 0.294, P = 0.009) and we found no one with 4977-bp deleted mitochondria. JC-1 (dye used as an indicator of MMP) ratio in the old group was significantly lower than the young group (3.01 ± 0.21 vs 3.85 ± 0.27, P = 0.033). Intracellular ROS levels between the groups did not differ significantly (P = 0.191). The intracellular ATP level in the young group was 1.75-fold higher than that of the advanced-age group (7.17 ± 1.16 vs 4.15 ± 0.60, P = 0.025). The protein expression of ATP5A1, as one of five proteins of OXPHOS, decreased with aging (P < 0.001). ATP5A1 mRNA expression was negatively correlated with aging (r = -0.341, P = 0.012).