汉防己甲素对先天性膈疝大鼠模型肺泡Ⅰ型上皮的影响  

Effects of Tetrandrine Prenatal Intervention on Alveolar Epithelial Cells Type Ⅰ Differentiation in Rat Model of Nitrofen-induced Congenital Diaphragmatic Hernia

作　　者：肖斌[1] 徐畅[1] 刘敏[1] 吉毅[1] 杨立勋[1] 李太明[1] 江君[1] 何涛贞[1] 

机构地区：[1]四川大学华西医院小儿外科,成都610041

出　　处：《四川大学学报（医学版）》2016年第2期169-173,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家青年自然科学基金(No.31201095)资助

摘　　要：目的探讨先天性膈疝(CDH)大鼠肺泡Ⅰ型上皮细胞的分化情况及汉防己甲素产前干预对其分化的影响。方法将孕9.5d的SD大鼠随机分为3组(对照组、膈疝组和治疗组),分别给予膈疝组和治疗组除草醚(以125mg/只的剂量一次性灌胃),对照组等量花生油。孕18.5d,分别给予治疗组汉防己甲素(灌胃给药30mg/kg,1次/d,连续3d),膈疝组和对照组等量生理盐水。孕21.5d,剖宫产取出胎肺,分别行HE染色并检测各组肺泡面积比,免疫组化检测甲状腺转录因子1(TTF1)蛋白的表达,实时荧光定量PCR(real-time PCR)检测RT140、TTF1mRNA的表达,电镜检测各组肺内Ⅰ型上皮细胞/Ⅱ型上皮细胞百分比(Ⅰ/Ⅱ%)。结果光镜和电镜下膈疝组胎肺明显发育不良,Ⅰ型上皮减少、Ⅱ型上皮增多,治疗组明显改善,且肺泡面积比呈对照组>治疗组>膈疝组(P<0.01);透射电镜提示Ⅰ/Ⅱ%呈对照组>治疗组>膈疝组(P<0.01);免疫组化及RT-PCR提示:TTF1表达呈对照组<治疗组<膈疝组(P<0.01),RT140表达呈对照组>治疗组>膈疝组(P<0.01)。结论先天性膈疝大鼠肺内存在肺泡Ⅰ型上皮分化障碍,汉防己甲素产前干预能促进CDH胎鼠肺发育。Objective To investigate the effects of Tetrandrine（TET）prenatal intervention on the differentiation of alveolar epithelial cells typeⅠ（AECⅠ）in rat model of Nitrofen-induced congenital diaphragmatic hernia（CDH）.Methods Timed-pregnant Sprague-Dawley rats were divided into three groups,namely control,CDH and TET group on day 9.5of gestation.The rats in TET group and CDH group were given 125 mg of Nitrofen by gavage one time,while the rats in control group were given the same dose of seed fat.After that,the rats in TET group was given 30 mg/kg of TET by gavage once a day for three days from day 18.5of gestation,while the rats in CDH and control group were given the same dose of normal saline.On day 21.5of gestation,all fetuses were delivered by cesarean,the lungs of fetuses were histologically evaluated by microscope and electron microscope.The expressions of typeⅠ cell-specific protein（RT140）and thyroid transcription factor 1（TTF1）in alveolar fluid content were analyzed by RT-PCR and immunohistochemistry staining.To detect the number of AECⅠ and AEC Ⅱ of each group by transmission electron microscopy and calculate the percentage of AEC Ⅰ and AEC Ⅱ（Ⅰ/Ⅱ %）.Results The microscope and electron microscope study found the lungs of fetuses in CDH group showed marked hypoplasia,in contrast to the improvement of hypoplasia in TET fetuses.The pulmonary alveolar area had significant difference statistically（P0.01）in each group,which present as controlTETCDH.Ⅰ /Ⅱ % had significant difference statistically（P0.01）in each group,which present as controlTETCDH.The expression level of TTF1 was up-regulated in both CDH and TET groups,and it was higher in CDH group（P0.01）.The expression level of RT140 were down-regulated in CDH and TET groups,which was lower in CDH group（P0.01）.Conclusion The development of AEC Ⅰ was interfered in CDH rat model,TET prenatal treatment could improve the lung development of CDH.

关 键 词：先天性膈疝 肺发育 Ⅰ型肺上皮细胞 汉防己甲素 

分 类 号：R726.5[医药卫生—儿科] R-332[医药卫生—临床医学]