寿胎丸对邻苯二甲酸二乙酯染毒孕鼠免疫功能及氧化应激的影响

Effects of Shoutai pills on immune function and oxidative stress in pregnant rats with di(2-ethylhexyl)phthalate exposure

目的探讨寿胎丸对邻苯二甲酸二乙酯(DEHP)染毒孕鼠免疫功能及氧化应激的影响。方法将36只性成熟雌性SD大鼠随机分为对照组、模型组、中药组,每组12只。确认妊娠后,分别予对照组玉米油10 mL/kg+生理盐水10 mL/kg、模型组DEHP500 mg/kg+生理盐水10 mL/kg、中药组DEHP 500 mg/kg+寿胎丸10 mL/kg灌胃,至妊娠第19天处死孕鼠。剖腹取出胎鼠,记录各组胎鼠体质量、孕鼠活胎数、死胎数;检测血清白介素-6(IL-6)、白介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)、雌二醇、孕激素水平;记录各组胎盘外观、色泽、质量情况,取胎盘组织做病理切片;检测胎盘组织中总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、活性氧(ROS)、丙二醛(MDA)水平。结果与对照组比较,模型组大鼠孕中后期体质量增加缓慢,胎鼠质量显著降低(P<0.05),血清IL-2、IL-6、TNF-α水平降低,雌二醇水平升高(P<0.05),胎盘T-AOC、GSH-Px、SOD、CAT水平显著降低,ROS、MDA水平显著升高(P<0.01)。与模型组比较,中药组大鼠孕中后期体质量明显增加,胎鼠质量增加(P<0.05);血清IL-2、IL-6升高,雌二醇降低(P<0.05),TNF-α有升高趋势(P>0.05),胎盘T-AOC、GSH-Px、CAT水平升高,MDA值明显降低(P<0.05),SOD升高,ROS降低(P>0.05)。孕激素各组皆无统计学差异(P>0.05)。HE染色结果显示,模型组胎盘组织海绵滋养层明显疏松杂乱,出现较多空泡。中药组胎盘组织结构完整,迷路带、海绵滋养层、巨细胞滋养层均清晰可见,各层中细胞分布均优于模型组。结论寿胎丸可调节孕鼠免疫功能,机制可能与其拮抗DEHP的雌激素样作用,调节血清免疫因子含量有关;寿胎丸可能通过纠正DEHP引起的胎盘组织内氧化应激失衡状态,减少胎盘组织损伤,改善妊娠结局。

Objective To investigate the effects of Shoutai pills(a traditional Chinese medicinal preparation)on immune functions and oxidative stress in pregnant rats exposed to di(2-ethylhexyl)phthalate(DEHP).Methods Thirty-six mature female SD rats were randomly divided into 3 groups(n=12).After pregnancy was confirmed,the rats were given 10 mL/kg corn oil+10 mL/kg saline(control group),500 mg/kg DEHP+10 mL/kg saline(model group),and 500 mg/kg DEHP+10 mL/kg Shoutai pills(treatment group).At 19 days of gestation,the rats were sacrificed and the fetal rats were weighed and the numbers of live and stillborn fetal rats were recorded.Serum levels of interleukin-6(IL-6),interleukin-2(IL-2),tumor necrosis factor-ɑ(TNF-ɑ),estradiol(E2)and progesterone(P)levels were detected.The appearance,color and quality of the placenta in each group were recorded,and the placental tissues were examined pathologically.The total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT),reactive oxygen species(ROS)and malondialdehyde(MDA)in the placental tissues were measured.Results Compared with the control group,the rats with DEHP exposure showed slow weight gain in the middle and late gestation period and significantly lower fetal weight(P<0.05)with lowered serum levels of IL-2,IL-6 and TNF-ɑ,increased estradiol level(P<0.05),decreased placental T-AOC,GSH-Px,SOD and CAT levels,and increased ROS and MDA levels(P<0.01).Compared with the model group,the rats treated with Shoutai pills had significantly increased weight gain in mid and late pregnancy and greater fetal weight(P<0.05)with significantly increased serum IL-2 and IL-6 levels,decreased estradiol level(P<0.05),slightly increased TNF-ɑexpression(P>0.05),increased placenta T-AOC,GSH-Px and CAT levels,decreased MDA level(P<0.05),and slightly increased SOD and decreased ROS levels(P>0.05).No significant difference was found in progesterone levels among the groups(P>0.05).HE staining showed that the trophoblast in the placental tissue sponge in the model group was loose and irregular with numerous vacuoles.In the treatment group,the structure of the placenta remained intact with clearly visible labyrinth zone,sponge trophoblast and giant cell trophoblast,and the cell distribution in each layer was better than that in the model group.Conclusion Shoutai pills can regulate the immune function of DEHP-exposed pregnant rats possibly by antagonizing the estrogenlike effect of DEHP and regulating serum immune factors;Shoutai pills can also reduce placental tissue damage and improve pregnancy outcome by correcting DEHP-induced imbalance of oxidative stress in the placental tissues.