Guizhi Fuling Wan inhibits autophagy of granulosa cells in polycystic ovary syndrome mice via H19/miR-29b-3p

Abstract

Objective

To investigate the potential molecular mechanism of traditional Chinese medicine Guizhi Fuling Wan (GZFLW) inhibiting granulosa cells (GCs) autophagy in polycystic ovary syndrome (PCOS).

Methods

Control GCs and model GCs were cultured and treated with blank serum or GZFLW-containing serum. The levels of H19 and miR-29b-3p in GCs were detected using qRT-PCR, target genes of miR-29b-3p were identified using luciferase assay. The protein expressions of Phosphatase and tensin homolog (PTEN), Matrix Metalloproteinase (MMP)-2, and Bax were measured using western blot. The level of autophagy was detected via MDC staining, the degree of autophagosomes and autophagic polymers was observed using dual fluorescence-tagged mRFP-eGFP-LC3.

Results

GZFLW intervention reduced the expression of autophagy-related proteins PTEN, MMP-2 and Bax, by upregulating the expression of miR-29b-3p and downregulated the expression of H19 (p < .05 or p < .01). The number of autophagosomes and autophagy polymers was significantly decreased by GZFLW treatment. However, the inhibition of miR-29b-3p and overexpression of H19 induced a significant increase in the number of autophagosomes and autophagic polymers, which attenuated the inhibitory effect of GZFLW on autophagy (p < .05 or p < .01). In addition, inhibition of miR-29b-3p or overexpression of H19 can attenuate the effect of GZFLW on the expression of PTEN, MMP-2 and Bax proteins (p < .05 or p < .01).

Conclusion

Our study found that GZFLW inhibits autophagy in PCOS GCs via H19/miR-29b-3p pathway.

Keywords:

• Guizhi Fuling Wan (GZFLW)
• polycystic ovary syndrome (PCOS)
• granulosa cell
• H19/miR-29b-3p

Disclosure statement

The authors report there is no competing interests to declare.

Author contributions

Hongqiu Zhu designed the experiments. Peijuan Wu, Ying Zhu, Xiaodan Hu, Haiyan Chen were performed experiments and collected data. Peijuan Wu discussed the results and strategy. Junjie Li, and Hanxue Wu supervised, directed and managed the study.

Additional information

Funding

This work was supported by National Natural Science Foundation of China (81873335), Science Technology Department of Sichuan Province (2020YJ0376).