孕鼠产前川芎嗪灌胃对先天性膈疝胎鼠肺组织发育及Hippo信号通路蛋白表达的影响  

Effects of prenatal tetramethylpyrazine intervention on lung development and Hippo signaling pathway protein expression in rat models of congenital diaphragmatic hernia

作　　者：廖君左[1] 蒋琳[2] 王城[1] 赵丹[1] LIAO Junzuo;JIANG Lin;WANG Cheng;ZHAO Dan(Department of Pediatric Surgery,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院小儿外科,四川南充637000 [2]川北医学院附属医院肿瘤科

出　　处：《山东医药》2023年第17期38-41,共4页Shandong Medical Journal

基　　金：四川省南充市市校战略合作专项(20SXQT0086);四川省卫计委科研课题普及应用项目(17PJ593)。

摘　　要：目的通过观察孕鼠产前川芎嗪灌胃对先天性膈疝胎鼠肺组织发育及Hippo信号通路蛋白表达的影响,初步探讨川芎嗪产前干预对先天性膈疝胎鼠的保护作用及其机制。方法SD大鼠按照雌雄2∶1的比例分笼夜间交配,于孕鼠妊娠9.5 d时随机分为正常对照组、模型对照组、模型+川芎嗪组各5只。模型对照组及模型+川芎嗪组在分组后立即给予除草醚一次性灌胃建立胎鼠膈疝模型,正常对照组给予等量橄榄油灌胃。模型+川芎嗪组从妊娠11.5 d开始给予80 mg/(kg·d)的川芎嗪一次性灌胃给药直至妊娠21 d,正常对照组、模型对照组给予等量蒸馏水灌胃。所有孕鼠于妊娠21.5 d行剖宫产取出胎鼠,取出胎鼠两侧肺组织观察有无膈疝,选择模型对照组及模型+川芎嗪组仅有左侧膈疝的胎鼠肺组织进行研究,取正常对照组胎鼠的左侧肺组织作为正常对照。HE染色及血管特殊染色观察各组肺组织形态和肺泡发育指标辐射状肺泡计数(RAC)、肺泡面积百分比、肺泡间隔厚度,以及肺血管发育指标血管外径、血管中膜百分比、血管壁肌化程度。Western blotting法检测各组肺组织Hippo信号通路相关蛋白Yap、Lats1、磷酸化Yap(p-Yap)表达。结果HE染色显示,与正常对照组比较,模型对照组胎鼠肺组织发育明显滞后,肺泡、肺泡囊及肺泡管均呈假腺体样结构,肺泡间隔及腺泡内动脉壁增厚;与模型对照组比较,川芎嗪干预组胎鼠肺组织发育有明显改善。模型对照组RAC、肺泡面积百分比小于正常对照组,肺泡间隔厚度大于正常对照组;模型+川芎嗪组RAC、肺泡面积百分比大于模型对照组,肺泡间隔厚度小于模型对照组(P均<0.05)。模型对照组血管中膜百分比高于正常对照组,血管壁未肌化程度低于正常对照组;模型+川芎嗪组血管中膜百分比低于模型对照组,血管壁未肌化程度高于模型对照组(P均<0.05)。模型对照组、模型Objective To observe the effects of prenatal intragastric administration of tetramethylpyrazine(TMP)on lung development and Hippo signaling protein expression in fetal rats with congenital diaphragmatic hernia(CDH),and to investigate the protective effect of TMP prenatal intervention on CDH fetal rats and its mechanism.Methods SD rats were mated at night according to the ratio of male and female 2∶1,and pregnant rats were randomly divided into the control group(n=5),CDH group(n=5)and CDH+TMP group(n=5).Nitrofen(125 mg/kg)was used to induce CDH on day 9.5 of pregnancy,and rats in the control group were given the equal volume of olive oil.TMP(80 mg/kg)was used as antenatal intervention on day 11.5 of pregnancy(once a day,for 10 days),while the rats in the control group and the CDH group were given the same amount of distilled water.All fetuses were delivered by cesarean on day 21.5 of pregnancy.The fetuses only with left diaphragmatic hernia in the CDH and CDH+TMP groups were chosen for analysis,while left lung tissues of the fetuses in the control group were taken as controls.HE staining and special blood vessel staining were used to observe lung tissue morphology,radiative alveolar count(RAC),alveolar area percentage,alveolar septal thickness,vascular outer diameter,vascular media percentage,and vascular wall muscle degree in each group.The expression levels of Hippo signaling pathway-related proteins Yap,Lats1 and phosphorylated Yap(p-Yap)in the lung tissues of each group were detected by Western blotting.Results Compared with the control group,the development of lung tissue in the CDH group was significantly delayed,the alveolar,alveolar sacs and alveolar ducts appeared pseudoglandular structures,and the alveolar septum and intracacinar artery wall were thickened.Compared with the CDH group,the development of lung tissue in the CDH+TMP group was significantly improved.The percentage of alveolar area and RAC in the CDH group was lower than that in the control group,and the thickness of alveolar septum was high

关 键 词：川芎嗪 肺发育不良 新生儿持续性肺动脉高压 Hippo信号通路 先天性膈疝 

分 类 号：R722.19[医药卫生—儿科]