摘要
目的观察邻苯二甲酸二(2-乙基已基)酯(DEHP)对妊娠期大鼠肺泡巨噬细胞(AM)内脂质过氧化反应及肺组织骨桥蛋白(OPN)表达的影响。方法 24只SD大鼠随机分为四组,每组6只。从孕第1天开始至子鼠出生后第21天,对照组(NC)给予玉米油0.5 mL/d,其余三组给予玉米油和10、100、750 mg/(kg.d)DEHP混合物0.5 mL灌胃染毒,分别为低、中、高剂量组。HE、Masson三色染色光镜下观察孕鼠肺组织的形态学改变,采用免疫组织化学染色法测定肺组织中的OPN。用含10%小牛血清的RPMI-1640细胞培养液分离、培养肺泡AM,并测定其超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GSH-PX)、一氧化氮合酶(NOS)的活性及丙二醛(MDA)、NO含量。结果高剂量组、中剂量组GSH-PX、SOD活性明显低于NC组,MDA、NO含量及NOS活性显著高于NC组(P均<0.05);HE染色高剂量组显示肺间质增厚、肺间质细胞增多,肺泡数目减少;Masson三色染色显示,低剂量组肺细小动脉以及肺间质均见有蓝色的胶原纤维沉积,中剂量组和高剂量蓝色胶原纤维沉积更为显著;免疫组化法显示NC组孕鼠肺组织OPN表达主要见于支气管、肺泡上皮细胞内;低剂量组大鼠肺细小动脉内皮细胞、平滑肌细胞以及周围巨噬细胞均见有OPN的表达,中剂量组和高剂量表达更强。结论 DEHP可使妊娠期大鼠肺泡AM内脂质过氧化反应增强,并促进肺组织内OPN过表达。
Objective To explore the effect of diethylhexylphthalate（DEHP） exposure during gestation and lactation on the function of alveolar macrophages and expression of osteopontin（OPN） in rats.Method 24 SD pregnant rats were randomly divided into 4 groups according to the different dosages of DEHP.Female rats were gavaged with 0（control group,n=6）,10 mg/kg（low dose group,n=6）,100 mg/kg（middle dose group,n=6） and 750 mg/kg（high dose group,n=6）DEHP respectively from gestational day（GD） 1 to postnatal day（PND） 21.The tissue sections of lung were stained with HE and Masson trichrome for light microscopic（LM） morphometric investigation.The expression of OPN protein in lung tissue was determined by immunohistochemistry alveolar macrophages were isolated and cultured with 10% Fetal Bovine Serum-RPMI-1640.Then,the activity of superoxide dismutase（SOD）,GSH-PX,malondialdehyde（MDA）,nitric oxide（NO） and nitric oxide synthase（NOS） in the macrophages were detected by spectophotometer respectively.Result Compared with normal control group,the activity of GSH-PX and SOD decreased,while the content of NO,NOS and MDA increased significantly in alveolar macrophages of the middle and high dose groups P0.05 all.In high dose group,alveolar septa were thicker,the number of cells increased in the alveolar septa,and the number of alveolar decreased inversely than those of normal control group（P0.05 all）.Collagen fiber was found both in the walls of blood vessels and in the alveolar septa for low dose group.Furthermore,more collagen fiber were observed in middle and high doses groups.Pulmonary arterioles,endothelial cells,macrophage,the walls of blood vessels and the alveolar septa show the protein expression of OPN in low dose group.Compared with the normal control group,immunohistochemistry analysis showed that protein expression of OPN was increased significantly in middle and high dose groups（P0.01）.Conclusion DEHP can enhance antilipid peroxidation level and inhibite OPN expression
出处
《山东医药》
CAS
2012年第6期11-13,共3页
Shandong Medical Journal
基金
温州市科技计划资助项目(Y20100130)
