从月经血中分离出的子宫内膜间充质干细胞通过抑制细胞周期通路中 Gadd45b 的表达来修复表阿霉素诱导的人卵巢颗粒细胞损伤。

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.

机构信息

Department of Obstetrics & Gynecology, General Hospital, Tianjin Medical University, NO.154, Anshan Road, Heping District, Tianjin, 300052, China.

Maternal and Child Health Care Hospital of Shandong Province, NO.238, Jingshi Road, Jinanlixia District, Jinan, 250014, China.

出版信息

Stem Cell Res Ther. 2019 Jan 3;10(1):4. doi: 10.1186/s13287-018-1101-0.

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.

背景

探讨间充质干细胞（MSC）对表柔比星诱导的人卵巢颗粒细胞（GC）损伤的作用及其潜在机制。

方法

采用酶联免疫吸附试验检测雌二醇、孕酮、抗苗勒管激素、抑制素 A 和抑制素 B 的水平。通过细胞计数试剂盒-8 检测 GC 的增殖。通过碘化丙啶单染检测细胞周期分布。通过 Annexin V 和 7-AAD 双染检测 GC 的凋亡。采用 Affymetrix Human Transcriptome Array 2.0 基因芯片分析 GC 的差异表达基因，并通过 Western blot 分析进行验证。

结果

表柔比星抑制雌二醇、孕酮、抗苗勒管激素、抑制素 A 和抑制素 B 的分泌和 GC 的增殖；将这些 GC 阻滞在 G2/M 期；并促进 GC 的凋亡。然而，MSC 修复了表柔比星对 GC 的损伤。通过微阵列和生物信息学分析发现 GC 的差异表达基因 Gadd45b、CyclinB1 和 CDC2。Western blot 显示，表柔比星上调 Gadd45b 蛋白表达，下调 CyclinB1 和 CDC2 蛋白表达，而 MSC 下调 Gadd45b 蛋白表达，上调 CyclinB1 和 CDC2 蛋白表达。