1.Regulation of osteopontin on decidualization of mouse endometrium in vitro
Yiyuan MEI ; Xiaobo WANG ; Qingzhen XIE
Chinese Journal of Reproduction and Contraception 2022;42(1):65-71
Objective:To explore whether osteopontin (OPN) is involved in the regulation of mouse endometrial stromal cells (mESCs) decidualization in vitro. Methods:Primary mESCs were treated with estrogen (10 nmol/L) and progesterone (1 μmol/L) to induce decidualization in vitro, and to detect the expression of OPN during mESCs decidualization. mESCs were transfected with over-expression vector of Opn, Opn shRNA, or with blank group as a negative control. The transfection efficiency was detected by real time PCR and Western blotting. The cell proliferation was detected by CCK-8 kit. Expressions of decidual/trophoblast-layer prolactin related protein (Dtprp), vascular endothelial growth factor A (Vegfa) and its receptor 2 (Vegfr2) were detected by real time PCR and Western blotting. Results:The mRNA levels of Dtprp ( P<0.001), Vegfa ( P=0.004) and Vegfr2 ( P=0.002) in decidualized mESCs were significantly increased, accompanied with the upregulated of Opn mRNA ( P=0.002) and OPN protein ( P<0.001) levels. Opn overexpression promoted mESCs proliferation ( P=0.004), while Opn knockdown can inhibit mESCs proliferation ( P=0.008). In addition, Opn overexpression further increased the mRNA levels of Dtprp ( P<0.001), Vegfa ( P=0.021), and Vegfr2 ( P=0.012) in decidualized mESCs, as well as the protein level of VEGFA ( P=0.001). Opn knockdown, on the other hand, inhibited the mRNA expression levels of Dtprp ( P=0.009), Vegfa ( P=0.007), and Vegfr2 ( P=0.001) in decidualized mESC, and the protein level of VEGFA ( P<0.001). Conclusion:OPN may play an important role in the regulation of mouse endometrial stromal cell decidualization.
2.Regulation of osteopontin on decidualization of mouse endometrium in vitro
Yiyuan MEI ; Xiaobo WANG ; Qingzhen XIE
Chinese Journal of Reproduction and Contraception 2022;42(1):65-71
Objective:To explore whether osteopontin (OPN) is involved in the regulation of mouse endometrial stromal cells (mESCs) decidualization in vitro. Methods:Primary mESCs were treated with estrogen (10 nmol/L) and progesterone (1 μmol/L) to induce decidualization in vitro, and to detect the expression of OPN during mESCs decidualization. mESCs were transfected with over-expression vector of Opn, Opn shRNA, or with blank group as a negative control. The transfection efficiency was detected by real time PCR and Western blotting. The cell proliferation was detected by CCK-8 kit. Expressions of decidual/trophoblast-layer prolactin related protein (Dtprp), vascular endothelial growth factor A (Vegfa) and its receptor 2 (Vegfr2) were detected by real time PCR and Western blotting. Results:The mRNA levels of Dtprp ( P<0.001), Vegfa ( P=0.004) and Vegfr2 ( P=0.002) in decidualized mESCs were significantly increased, accompanied with the upregulated of Opn mRNA ( P=0.002) and OPN protein ( P<0.001) levels. Opn overexpression promoted mESCs proliferation ( P=0.004), while Opn knockdown can inhibit mESCs proliferation ( P=0.008). In addition, Opn overexpression further increased the mRNA levels of Dtprp ( P<0.001), Vegfa ( P=0.021), and Vegfr2 ( P=0.012) in decidualized mESCs, as well as the protein level of VEGFA ( P=0.001). Opn knockdown, on the other hand, inhibited the mRNA expression levels of Dtprp ( P=0.009), Vegfa ( P=0.007), and Vegfr2 ( P=0.001) in decidualized mESC, and the protein level of VEGFA ( P<0.001). Conclusion:OPN may play an important role in the regulation of mouse endometrial stromal cell decidualization.
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