Endometrial mesenchymal stem cells isolated from menstrual blood repaired epirubicin-induced damage to human ovarian granulosa cells by inhibiting the expression of Gadd45b in cell cycle pathway

Abstract

BACKGROUND: To explore the effect of mesenchymal stem cells isolated from menstrual blood (MB-MSCs) on epirubicin-induced damage to human ovarian granulosa cells (GCs) and its potential mechanisms. METHODS: The estradiol, progesterone, anti-Müllerian hormone, inhibin A, and inhibin B levels were determined using enzyme-linked immunosorbent assay. The proliferation of GCs was detected by Cell Counting Kit-8 assays. The cell cycle distribution was detected by propidiumiodide single staining. The apoptosis of GCs was determined using Annexin V and 7-AAD double staining. The differentially expressed genes of GCs were analyzed with Affymetrix Human Transcriptome Array 2.0 gene chip and verified with Western blot analysis. RESULTS: Epirubicin inhibited the secretion of estradiol, progesterone, anti-Müllerian hormone, inhibin A, and inhibin B and the proliferation of GCs; arrested these GCs in G2/M phase; and promoted the apoptosis of GCs. However, MB-MSCs repaired epirubicin-induced damage to GCs. Differentially expressed genes of GCs, Gadd45b, CyclinB1, and CDC2, were found by microarray and bioinformatics analysis. Western blot showed that epirubicin upregulated Gadd45b protein expression and downregulated CyclinB1 and CDC2 protein expression, while MB-MSCs downregulated Gadd45b protein expression and upregulated CyclinB1 and CDC2 protein expression. CONCLUSIONS: MB-MSCs repaired epirubicin-induced damage to GCs, which might be related to the inhibition of Gadd45b protein expression.