视黄酸信号通路在布洛芬致小鼠卵巢损伤中的表达

Expression of retinoic acid signaling pathway in ibuprofen-induced ovarian injury in mice

目的 观察视黄酸(retinoic acid,RA)信号通路在布洛芬(ibuprofen, IBU)致小鼠卵巢损伤的表达,并分析其作用。方法 将7周龄ICR小鼠随机分为低、中、高剂量组和对照组,每组各14只,低、中和高剂量组连续7 d经口分别给予50、100和200 mg/kg·dIBU,对照组给予等体积生理盐水。停止给药1周后处死小鼠、称体重、取双侧卵巢并称其重量、计算卵巢脏器系数。苏木素-伊红(HE)染色后观察卵巢组织学改变,ELISA法检测血清雌二醇(estradiol, E2)水平,qRT-PCR方法检测RA信号通路相关基因的表达丰度,Western blot检测ALDH2和CYP26A1蛋白的表达水平。结果 高剂量组小鼠体重(29.362±1.035) g低于对照组(31.775±1.640) g,差异有统计学意义(P<0.05);随着IBU剂量增大,卵巢组织更加松散、颗粒细胞排列更加紊乱。中和高剂量组小鼠血清E2浓度分别为(47.89±3.44) pg/ml和(22.33±5.44) pg/ml,均低于对照组(58.52±12.58) pg/ml,差异有统计学意义(P<0.05);中、高剂量组卵巢组织中的合成视黄酸的Aldh2、Aldh1a1、Aldh1a2和Aldh1a3基因丰度高于对照组,高剂量组卵巢组织中视黄酸核受体(Rar和Rxr)相关基因丰度高于对照组,中、高剂量卵巢组织中代谢视黄酸的Cyp26a1、Cyp26b1和Cyp26c1基因丰度低于对照组,中、高剂量组卵巢组织凋亡基因Jnk家族丰度高于对照组,差异均有统计学意义(P<0.05);各实验组卵巢组织中参与RA合成的蛋白ALDH2表达高于对照组,各实验组卵巢组织中参与RA代谢的蛋白CYP26A1表达低于对照组,差异均有统计学意义(P<0.05)。结论 中、高剂量IBU可能使卵巢内RA合成和代谢改变而使其受到损伤。

Objective To observe the expression of retinoic acid(RA) signaling pathway in ibuprofen(IBU)-induced ovarian injury in mice, and to analyze its effect. Methods The 7-week-old ICR mice were randomly divided into low-dose, medium-dose and high-dose groups and control group, with 14 mice in each group. The low-dose, medium-dose and high-dose groups were given 50, 100 and 200 mg/kg·d IBU orally for 7 days, respectively, while the control group was given the same volume of normal saline. One week after the last administration, the mice were sacrificed, the body weight was weighed, the bilateral ovaries were taken and weighed, and the ovarian organ coefficient was calculated. The serum estradiol(E2) level was detected by ELISA, the expression abundance of RA signaling pathway related genes was detected by qRT-PCR, and the protein expression levels of ALDH2 and CYP26 A1 were detected by Western blot. Results The weight of mice in the high-dose group(29.362±1.035 g) was lower than that in the control group(31.775±1.640 g), and the difference was statistically significant(P<0.05). With the increase of IBU dose, ovarian tissue became looser and granulogranular cells became more disorganized. The serum E2 concentrations of mice in medium and high dose groups were(47.89±3.44 pg/ml) and(22.33±5.44 pg/ml), respectively, which were lower than those in control group(58.52±12.58 pg/ml), and the differences were statistically significant(P <0.05). The abundance of Aldh2, Aldh1a1, Aldh1a2 and Aldh1a3 genes for synthesis of retinoic acid in ovarian tissues of medium and high dose groups was higher than that of control group, and the abundance of retinoic acid nuclear receptor(Rar and Rxr) related genes in ovarian tissues of high dose group was higher than that of control group. The abundance of Cyp26a1, Cyp26b1 and Cyp26c1 genes metabolized retinoic acid in medium and high dose ovarian tissues was lower than that in the control group, and the abundance of Jnk family apoptotic genes in medium and high dose ovarian tissues was higher than that in the control group, and the differences were statistically significant(P<0.05). The expression of ALDH2 protein involved in RA synthesis in ovarian tissues of all experimental groups was higher than that of the control group, and the expression of CYP26 A1 protein involved in RA metabolism in ovarian tissues of all experimental groups was lower than that of the control group, and the differences were statistically significant(P<0.05). Conclusion Medium and high doses of IBU may cause changes in RA synthesis and metabolism in the ovary, resulting in damage to RA.