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Acbp is essential for decidualization during early pregnancy in mice
in Reproduction*(X Zhang and B-Y Tan contributed equally to this work)
Decidualization of uterine stromal cells plays an important role in the establishment of normal pregnancy. Previous studies have demonstrated that Acyl-CoA binding protein (Acbp) is critical to cellular proliferation, differentiation, mitochondrial functions, and autophagy. The characterization and physiological function of Acbp during decidualization remain largely unknown. In the present study, we conducted the expression profile of Acbp in the endometrium of early pregnant mice. With the occurrence of decidualization, the expression of Acbp gradually increased. Similarly, Acbp expression was also strongly expressed in decidualized cells following artificial decidualization, both in vivo and in vitro. We applied the mice pseudopregnancy model to reveal that the expression of Acbp in the endometrium of early pregnant mice was not induced by embryonic signaling. Moreover, P4 significantly upregulated the expression of Acbp, whereas E2 appeared to have no regulating effect on Acbp expression in uterine stromal cells. Concurrently, we found that interfering with Acbp attenuated decidualization, and that might due to mitochondrial dysfunctions and the inhibition of fatty acid oxidation. The level of autophagy was increased after knocking down Acbp. During induced decidualization, the expression of ACBP was decreased with the treatment of rapamycin (an autophagy inducer), while increased with the addition of Chloroquine (an autophagy inhibitor). Our work suggests that Acbp plays an essential role in the proliferation and differentiation of stromal cells during decidualization through regulating mitochondrial functions, fatty acid oxidation, and autophagy.
Supplementary Materials
- Supplementary Figure 1 Identification of mouse endometrial stromal cells (mESCs) based on the immunofluorescence. Identification of mESCs based on positive immunofluorescence staining by the anti-vimentin rabbit polyclonal antibody. Scale bar, 100 µm. The experiment was repeated three times.
- Supplementary Figure 2 Transfection efficiency of siRNA specifically targeting mouse Acbp (si-acbp) in mouse endometrial stromal cells (mESCs). (A) The Acbp-knockdown efficiency was shown by Western blot analysis. (B) The quantification of three western blot results. (C) The Acbp-knockdown efficiency was shown by RT-qPCR. ***p < 0.001. β-actin (ACTB) was used as an internal control. Each experiment was repeated three times.
- upplementary Figure 3 Selection of the concentration of CPT1 inhibitor etomoxir in mESCs. (A) Western blot detection of CPT1 protein expression at different concentrations (12.5μM, 25μM, 50μM and 100μM) of etomoxir. (B) Selection of the concentration of etomoxir by CCK8 analysis. ns，no significance. **p < 0.01 and ***p < 0.001. β-actin (ACTB) was used as an internal control. Each experiment was repeated three times.
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