汉防己甲素对先天性膈疝大鼠的保护作用

Protective effects of tetrandrine against congenital diaphragmatic hernia in rats

摘要:

目的：探讨汉防己甲素（TET）产前干预对先天性膈疝（CDH）大鼠的保护作用及机制。方法：将妊娠Sprague-Dawley大鼠随机分为对照组、除草醚组与TET治疗组。后两组孕 9.5 d 时采用除草醚灌胃法建立 CDH 大鼠模型；治疗组自孕18.5 d 起给予 TET 灌胃（每日30 mg/kg，连续3 d）；21 d 对部分孕鼠行剖腹并抽取羊水，观察胎鼠膈疝形成情况。采用 ELISA 法、免疫组化染色法检测羊水和胎肺中 TNF-α 的表达情况。余孕鼠自然分娩，观察各组仔鼠出生后情况。结果：除草醚组胎鼠无论有无膈疝形成均存在肺发育不良，肺及羊水中 TNF-α 的表达均明显升高；TET治疗组胎鼠巨大膈疝的发生率低于除草醚组，肺与羊水中 TNF-α的含量明显较除草醚组少（P＜0.01）。在自然分娩的仔鼠中，TET治疗组仔鼠的 24 h存活率明显高于除草醚组（P＜0.01）。结论：产前应用 TET 能降低CDH大鼠模型胎肺与羊水中 TNF-α 的含量，改善因除草醚诱导的胎鼠肺发育不良，减少巨大膈疝的发生，提高仔鼠的存活率。

Abstract:

OBJECTIVE: To study the protective effect of the prenatal use of tetrandrine (TET) against congenital diaphragmatic hernia (CDH) in rats and possible mechanisms. METHODS: Pregnant female Sprague-Dawley rats were randomly divided into 3 groups: control, nitrofen and TET treatment. The later two groups were administered with nitrofen by gavage on day 9.5 of gestation. On day 18.5 of gestation, TET (30 mg/kg) was given by gavage (once a day, for three days) in the TET treatment group. On day 21 of gestation, parts of pregnant rats were delivered by cesarean section and amniotic fluid was collected. The fetal rats were examined for a diaphragmatic hernia. Lung histologic evaluations with microscope and immunohistochemistry staining of TNF-α were performed. TNF-α in amniotic fluid was detected using ELISA. The remaining pregnant rats were allowed to deliver spontaneously at term. The survival of pup rats was observed until 24 hrs of age.ResultsIn the nitrofen group, significant lung hypoplasia was presented not only in fetuses with CDH but also in those without CDH. Stronger expression of TNF-α was observed in fetal lungs and amniotic fluid in the nitrofen group, even when CDH was absent. The TET treatment group showed improved lung development compared with the nitrofen group. The incidence of large diaphragmatic hernia in the TET treatment group was lower than that in the nitrofen group (P<0.05), and the expression of TNF-α in fetal lungs and amniotic fluid in the TET treatment group was also lower than in the nitrofen group (P<0.01). The 24-hr survival rate of pup rats in the TET group was higher than that in the ntrofen group (P<0.01). CONCLUSIONS: Prenatal use of TET can improve nitrofen-induced pulmonary hypoplasia, decrease the incidence of large diaphragmatic hernia and increase the survival rate of pup rats, possibly through a reduction in the production of TNF-α in fetal lungs and amniotic fluid in rats with CDH.