[粉防己碱产前干预对硝基芬诱导的先天性膈疝大鼠模型中Ⅰ型肺泡上皮细胞分化的影响]

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

作者信息

Xiao Bin, Xu Chang, Liu Min, Ji Yi, Li Tai-ming, Jiang Jun, He Tao-zhen

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2016 Mar;47(2):169-73.

Abstract

OBJECTIVE

To investigate the effects of Tetrandrine (TET) prenatal intervention on the differentiation of alveolar epithelial cells type I (AEC I) 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 9.5 of 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 18.5 of gestation, while the rats in CDH and control group were given the same dose of normal saline. On day 21.5 of gestation, all fetuses were delivered by cesarean, the lungs of fetuses were histologically evaluated by microscope and electron microscope. The expressions of type I 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 AEC I 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 (P < 0.01) in each group, which present as control > TET > CDH. I/II% had significant difference statistically (P < 0.01) in each group, which present as control > TET > CDH. The expression level of TTF1 was up-regulated in both CDH and TET groups, and it was higher in CDH group (P < 0.01). The expression level of RT140 were down-regulated in CDH and TET groups, which was lower in CDH group (P < 0.01).

CONCLUSION

The development of AEC I was interfered in CDH rat model, TET prenatal treatment could improve the lung development of CDH.

摘要

目的

探讨粉防己碱（TET）产前干预对硝基芬诱导的先天性膈疝（CDH）大鼠模型中I型肺泡上皮细胞（AEC I）分化的影响。

方法

将孕龄合适的Sprague-Dawley大鼠在妊娠第9.5天分为三组，即对照组、CDH组和TET组。TET组和CDH组大鼠一次性灌胃给予125 mg硝基芬，而对照组大鼠给予相同剂量的种子油。之后，TET组大鼠从妊娠第18.5天开始每天一次灌胃给予30 mg/kg TET，连续三天，而CDH组和对照组大鼠给予相同剂量的生理盐水。在妊娠第21.5天，所有胎儿通过剖宫产取出，胎儿的肺组织通过显微镜和电子显微镜进行组织学评估。通过RT-PCR和免疫组织化学染色分析肺泡液中I型细胞特异性蛋白（RT140）和甲状腺转录因子1（TTF1）的表达。通过透射电子显微镜检测每组AEC I和AEC II的数量，并计算AEC I和AEC II的百分比（I/II%）。

结果

显微镜和电子显微镜研究发现，CDH组胎儿的肺显示出明显的发育不全，而TET组胎儿的发育不全有所改善。每组肺泡面积在统计学上有显著差异（P < 0.01），表现为对照组>TET组>CDH组。I/II%在每组中在统计学上有显著差异（P < 0.01），表现为对照组>TET组>CDH组。CDH组和TET组中TTF1的表达水平均上调，且CDH组更高（P < 0.01）。CDH组和TET组中RT140的表达水平均下调，且CDH组更低（P < 0.01）。