Mao Zenghui, Fan Liqing, Yu Qiao, Luo Shuwei, Wu Xianling, Tang Jun, Kang Gehua, Tang Le

1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 2 Institute of Reproduction and Stem Cell Engineering, Central South University, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.| 1 Reproductive Medicine Center, Reproductive Medicine Hospital of Hunan Province, Changsha, Hunan, People's Republic of China.

Reprod Sci. 2018 Mar;25(3):372-383. doi: 10.1177/1933719117715129. Epub 2017 Jul

This study investigated the involvement of the klotho-associated signaling in the apoptosis of granulosa cells (GCs) from the ovaries of patients with polycystic ovary syndrome (PCOS) and PCOS animals. Primary GCs were obtained from 26 healthy women and 43 women with PCOS. The PCOS animal model was established by the injection of dehydroepiandrosterone (DHEA). Klotho protein and associated microRNA expression in human primary GCs and rats' ovarian tissues were measured by Western blot and real-time polymerase chain reaction, respectively. Results showed that significantly lower miR-126-5p and miR-29a-5p microRNA expressions, higher klotho protein expression, lower insulin growth factor 1 (IGF-1R) and Wnt family member 1 (Wnt1) protein expressions, and lower Akt phosphorylation at Ser(473) and Thr(308) residues were observed in the GCs from patients with PCOS and the ovarian tissues of PCOS rats compared to that in GCs from healthy women and ovarian tissues of normal control rats, respectively. Knockdown of klotho gene expression normalized IGF-1R and Wnt1 protein expressions and Akt phosphorylation in GCs from patients with PCOS and the ovarian tissues from PCOS rats; it also blocked the effects of insulin on apoptosis and proliferation in GCs from patients with PCOS and inhibited caspase-3 activity in ovarian tissues of PCOS rats. Knockdown of klotho gene expression increased the pregnancy rate in DHEA-treated female rats and increased the body weight of their newborns through normalizing the ovarian function and decreasing the formation of cystic follicles. In conclusion, the miR-126-5p, miR-29a-5p/klotho/insulin-IGF-1, Wnt, and Akt signal pathway may be involved in the apoptosis of GCs and subsequent development of PCOS.

Ding Cai-Fei, Chen Wang-Qiang, Zhu Yu-Tian, Bo Ya-Li, Hu Hui-Min, Zheng Ruo-Heng

Reproductive Department, Integrated Chinese and Western Medicine Hospital of Zhejiang Province , Hangzhou , P. R. China.

Hum Fertil (Camb). 2015 Mar;18(1):22-9. doi: 10.3109/14647273.2014.956811. Epub

AIM: To explore the pattern of expression of circulating miRNAs in patients with polycystic ovary syndrome (PCOS). MATERIALS AND
METHODS: Microarray and qRT-PCR were used to investigate circulating miRNAs in PCOS during clinical diagnosis. The targets of dys-regulated miRNAs were predicted using bioinformatics, followed by function and pathway analysis using the databases of Gene Ontology and the KEGG pathway.
RESULTS: BMI, triglyceride, HOMA-IR, Testosterone and CRP levels were significantly higher, while estradiol was significantly lower in PCOS than in control groups. After SAM analysis, 5 circulating miRNAs were significantly up-regulated (let-7i-3pm, miR-5706, miR-4463, miR-3665, miR-638) and 4 (miR-124-3p, miR-128, miR-29a-3p, let-7c) were down-regulated in PCOS patients. Hierarchical clustering showed a general distinction between PCOS and control samples in a heat map. After joint prediction by different statistical methods, 34 and 41 genes targeted were up-and down-regulated miRNAs, in PCOS and controls, respectively. Further, GO and KEGG analyses revealed the involvement of the immune system, ATP binding, MAPK signaling, apoptosis, angiogenesis, response to reactive oxygen species and p53 signaling pathways in PCOS.
CONCLUSIONS: We report a novel non-invasive miRNA profile which distinguishes PCOS patients from healthy controls. The miRNA-target database may provide a novel understanding of PCOS and potential therapeutic targets.