Shen Yan-Qing, Yang Yang, Sun Zhong-Yi, Li Shu-Jun, Shen Jin-Xin, Zhou Xiao-Yu
Neonatal Medical Center, Children's Hospital Affiliated to Nanjing Medical University, Nanjing 210008, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2019 Mar;21(3):287-293. doi: 10.7499/j.issn.1008-8830.2019.03.019.
To study the role of miR-431 in lung development and morphology.
According to the stage of lung development in rats, Sprague-Dawley rats at embryonic day 16 (E16), embryonic day (E19), embryonic day (E21), postnatal day 1 (P1), postnatal day 3 (P3), postnatal day 7 (P7), postnatal day 14 (P14) and 10 weeks after birth (P10 weeks) were selected, and lung tissue samples were collected for observation. Hematoxylin-eosin staining and transmission electron microscopy were performed to observe the morphology of lung tissue. Fluorescence in situ hybridization and real-time PCR were used to measure the expression of miR-431 during the critical stages of lung development (E19, E21 and P3).
The E19 group had the formation of the lamellar body and type II alveolar epithelial cells in the fetal lung tissue. The number of lamellar bodies increased with the increasing gestational age, with aggregation and excretion. Pulmonary alveoli formed rapidly, the lung interstitium became thinner, and the microvascular system became mature after birth. Fluorescence in situ hybridization and real-time PCR showed that the expression of miR-431 gradually decreased with the increasing gestational age (P<0.05).
The systematic and continuous morphological data of lung development is obtained in this experiment. In addition, miR-431 may play an important role in the negative regulation of lung development, which provides basis and direction for further research on the mechanism of lung development and related diseases.
研究miR-431在肺发育及形态学中的作用。
根据大鼠肺发育阶段,选取胚胎第16天(E16)、胚胎第19天(E19)、胚胎第21天(E21)、出生后第1天(P1)、出生后第3天(P3)、出生后第7天(P7)、出生后第14天(P14)及出生后10周(P10周)的Sprague-Dawley大鼠,采集肺组织样本进行观察。采用苏木精-伊红染色及透射电子显微镜观察肺组织形态。运用荧光原位杂交及实时定量PCR检测肺发育关键阶段(E19、E21及P3)miR-431的表达。
E19组胎儿肺组织出现板层小体及II型肺泡上皮细胞。板层小体数量随胎龄增加而增多,并伴有聚集及排泄。出生后肺泡迅速形成,肺间质变薄,微血管系统成熟。荧光原位杂交及实时定量PCR结果显示,miR-431表达随胎龄增加逐渐降低(P<0.05)。
本实验获得了肺发育系统且连续的形态学数据。此外,miR-431可能在肺发育的负调控中发挥重要作用,为进一步研究肺发育机制及相关疾病提供了依据和方向。