Aroclor 1254致孕鼠子宫内膜细胞损伤模型的建立

Establishment of injured endometrial cell model in pregnant rats induced by Aroclor 1254 in vitro

为建立Aroclor 1254致孕鼠子宫内膜细胞损伤模型,通过分离、培养怀孕大鼠的原代子宫内膜细胞,用不同浓度的Aroclor 1254处理子宫内膜细胞48 h,倒置显微镜下观察细胞形态,MTT法检测细胞的活力,Western blot法检测细胞中CYP450的表达,并通过ELISA法检测细胞培养基中TNF-α、IL-6、雌二醇(estradiol,E2)和孕酮(progesterone,P4)的含量,来确定Aroclor 1254是否对细胞造成损伤。结果表明,随着Aroclor 1254浓度的升高,子宫内膜细胞的损伤及抑制率也随之增高,CYP450的表达随Aroclor 1254浓度的升高而升高。TNF-α和IL-6的水平比对照组显著升高(P<0.05),E2和P4的水平则比对照组显著降低(P<0.05)。这表明Aroclor 1254对体外培养子宫内膜细胞损伤程度呈浓度依赖性,10 mg/L的质量浓度作用48 h可建立可靠的子宫内膜细胞损伤模型。

To establish the injured endometrial cell model in pregnant rats induced by Aroclor 1254,the primarily cultured endometrial cells in pregnant rats were exposed to different concentrations of Aroclor1254 for 48 h culture. The configuration of endometrial cells was detected under inverted microscope. The endometrial cells viability was determined by MTT assay and the expressions of CYP1A1 and CYP2B1 were determined by Western blot methods. The contents of TNF-α,IL-6,estradiol( E2) and progesterone( P4) in the supernatant of the endometrial cells were measured by the enzyme-linked immunosorbent assay( ELISA) method. It showed that,the inhibition rate of the cultured endometrial cells increased with the increase of Aroclor 1254 concentrations. The expressions of CYP1A1 and CYP2B1 increased with the treatment of the gradient Aroclor 1254. The contents of TNF-α and IL-6 in the injured endometrial cells were higher significantly than those of the control group( P < 0. 05),E2 and P4were lower significantly than those of the control group( P < 0. 05). It shows that the damage to endometrial cells induced by Aroclor1254 is dose-dependent. The injured endometrial cells model can be established successfully with the treatment of 10 mg / L Aroclor 1254 for 48 h.