邻苯二甲酸二乙基已基酯联合高氧通过调控Akt/GSK-3β信号通路介导细胞凋亡影响新生大鼠肺发育  

DEHP combined with hyperoxia arrests lung development of neonatal rats via Akt/GSK-3β and Bcl-2/Bax signaling pathway mediated apoptosis

作　　者：潘冰婷 俞彬媛 王芯叶 霍冉 梁忠杰[1] 陈尚勤[1] PAN Bing-ting;YU Bin-yuan;WANG Xin-ye;HUO Ran;LIANG Zhong-jie;CHEN Shang-qin(Department of Neonatology,the Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院新生儿科,浙江温州325027

出　　处：《中国药理学与毒理学杂志》2023年第3期178-184,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：浙江省自然科学基金(LY21H040007);温州市科技局基金(Y20180006)。

摘　　要：目的 探讨邻苯二甲酸二乙基己基酯(DEHP)联合高氧对新生大鼠肺发育的影响及其机制。方法新生SD大鼠随机分为4组:正常对照、高氧、DEHP和高氧+DEHP组。正常对照组大鼠置空气环境并每天ig给予玉米油5 mL·kg^(-1);DEHP组大鼠出生后每天ig给予DEHP 750 mg·kg^(-1);高氧组大鼠出生后12 h内即置氧箱中持续吸入80%~85%O2;高氧+DEHP组大鼠出生后12 h内即置于氧箱中并每天ig给予DEHP750 mg·kg^(-1)。各组出生后第7天取肺组织,HE染色观察肺组织形态改变,TUNEL染色观察肺上皮细胞凋亡,Western印迹法测定肺组织蛋白激酶B(Akt),糖原合成酶激酶3β(GSK-3β),Bcl-2,Bax和活化胱天蛋白酶3蛋白表达水平。结果 HE染色结果显示,DEHP、高氧及高氧+DEHP组肺泡结构均呈现不同程度简单化和肺泡数量减少的形态改变。TUNEL结果显示,与正常对照组相比,DEHP和高氧组细胞凋亡显著增加(P<0.01);与高氧组相比,高氧+DEHP组细胞凋亡显著增加(P<0.05)。Western印迹结果显示,与正常对照组相比,DEHP和高氧组肺组织中Akt和GSK-3β磷酸化水平及Bcl-2蛋白表达水平降低(P<0.05),Bax和活化胱天蛋白酶3表达水平增加(P<0.05);与高氧组相比,高氧+DEHP组肺组织中Akt和GSK-3β磷酸化水平及Bcl-2蛋白表达降低(P<0.05),Bax和活化胱天蛋白酶3表达增加(P<0.05)。结论 DEHP可阻滞新生大鼠肺发育,并加重或协同高氧致肺损伤,通过调控Akt/GSK-3β信号通路诱导细胞凋亡影响新生大鼠肺发育。OBJECTIVE To investigate the effect of di(2-ethylhexyl)phthalate(DEHP)combined with hyperoxia on lung development of neonatal rats and its mechanism.METHODS Newborn Sprague-Dawley(SD)rats were randomly divided into four groups:control,DEHP,hyperoxia and hyperoxia+DEHP.The control group was maintained in room air and received an equal volume of the vehicle.The DEHP group was treated with DEHP(750 mg·kg-1·d-1)by gavage on postnatal day 1-7.The hyperoxia group was exposed to hyperoxia(O280%-85%)in a sealed Plexiglas tank within 12 h of birth.The hyperoxia+DEHP group was exposed to hyperoxia(O280%-85%)in a sealed Plexiglas tank within 12 h of birth and treated with DEHP(750 mg·kg-1·d-1)by gavage on postnatal day 1-7.Lung tissues were collected on postnatal day 7 and morphological changes in the lungs were examined by hematoxylin and eosin staining(HE).Apoptosis in lung tissues was assessed by TUNEL staining.The protein expression levels of phosphorylated protein kinase B(p-Akt),Akt,glycogen synthase kinase-3β(GSK-3β),p-GSK-3β,Bcl-2,Bax and cleaved-caspase 3 were detected via Western blotting.RESULTS The HE results demonstrated that DEHP,hyperoxia and hyperoxia+DEHP groups all showed varying degrees of simplification of the alveolar structure and reduced numbers of alveoli.The TUNEL staining showed that the rate of apoptosis in lung tissues of DEHP and hyperoxia groups significantly increased compared with the control group(P<0.01),and the Bax and cleaved-caspase 3 expressions were increased(P<0.05).The case was the same with the hyperoxia+DEHP group compared with the hyperoxia group(P<0.05).Western blotting indicated that the ratio of p-Akt/Akt,p-GSK-3β/GSK-3βand the Bcl-2 expression in lung tissues of DEHP and hyperoxia groups were decreased compared with the control group(P<0.05),and the Bax and cleaved-caspase 3 expressions were increased(P<0.05).The case was the same with the hyperoxia+DEHP group compared with the hyperoxia group(P<0.05).CONCLUSION DEHP can arrest lung development of neonatal rats and ag

关 键 词：邻苯二甲酸二乙基己基酯 肺发育 蛋酶激酶B 糖原合成酶激酶3Β 细胞凋亡 

分 类 号：R994.6[医药卫生—毒理学]