槲皮素对Aroclor 1254致孕鼠子宫内膜细胞损伤的保护作用

Protective effects of quercetin on Aroclor 1254 induced endometrial cell injury in pregnant rats

为了研究槲皮素对Aroclor 1254损伤的孕鼠子宫内膜细胞是否具有保护作用,试验通过分离、培养怀孕大鼠的子宫内膜细胞,以Aroclor 1254诱导子宫内膜细胞损伤模型,用不同剂量的槲皮素分别处理损伤的子宫内膜细胞24~72h,MTT法测细胞的活力,RT-PCR及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,来确定槲皮素是否对损伤的细胞具有保护作用。结果显示,处理24h后,随着槲皮素浓度的升高,子宫内膜细胞的成活率也随之增高,其中50μmol/L槲皮素时成活率最高。CYP1A1、CYP2B1和CYP2E1在子宫内膜细胞mRNA中的表达随槲皮素浓度的升高呈上升的趋势。CYP1A1和CYP2E1在细胞的蛋白中不表达,CYP2B1的表达随槲皮素浓度的增加而呈升高的趋势。50μmol/L槲皮素作用24h对损伤的怀孕大鼠子宫内膜细胞CYP1A1、CYP2B1和CYP2E1的表达效果最明显。Aroclor 1254损伤的子宫内膜细胞中TNF-α和IL-6的含量比对照组显著升高(P<0.05),E2和P4的含量则比对照组显著降低(P<0.05)。50μmol/L槲皮素处理后TNF-α和IL-6的含量比Aroclor 1254损伤组显著降低(P<0.05),E2和P4则显著升高(P<0.05)。这表明槲皮素对损伤的怀孕大鼠子宫内膜细胞具有保护作用。

The objective of this study was to investigate the protective effects of quercetin on Aroclor 1254 damaged to endometrial cells in pregnant rats.The primary cultured endometrial cells were isolated and Aroclor 1254 was used to induce the injured endometrial cells model.The cells were treated with gradient quercetin for 24,48 hand 72hrespectively,the viability of the endometrial cells was measured by MTT method,and the expressions of CYP1A1,CYP 2B1 and CYP2E1were determined by RT-PCR and 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 to determine whether the quercetin plays the protective role in the damaged cells.It showed that,the survival rate of the cultured endometrial cells increased with the increase of quercetin concentrations starting from the time point 24 h post culture.The highest survival rate of cultured endometrial cells occurred in the 50μmol/L quercetin.The expressions of CYP1A1,CYP2B1 and CYP2E1in mRNA of the endometrial cells increased gradually with the treatment of the gradient quercetin for 24 h.In the Western blot method,CYP2B1 expression increased gradually with the treatment of the gradient quercetin for24 h,there were no expression of CYP1A1 and CYP2E1.The optimal quercetin for CYP1A1,CYP2B1 and CYP2E1expression was 50μmol/L for 24 h.The contents of TNF-αand IL-6in the injured endometrial cells were significantly higher than those in the control group(P<0.05),E2 and P4 were significantly lower than those in the control group(P<0.05).By the treatment of quercetin,the contents of TNF-αand IL-6 were significantly lower than those in Aroclor 1254group(P<0.05),E2 and P4increased significantly.It shows that quercetin has protective effects on the injured endometrial cells in the pregnant rats.