不同剂量 rhEPO 对新型支气管肺发育不良大鼠肺eNOS和Bcl-2蛋白表达的影响

摘要

目的：观察使用不同治疗剂量的重组人促红细胞生成素（rhEPO）对SD新生幼鼠的新型支气管肺发育不良（BPD）模型中内皮型一氧化氮合酶（eNOS）、Bcl‐2在肺组织的表达水平变化的影响，并找出合适的给药时间及使用剂量，旨在探讨其对新型BPD的保护作用。方法利用脂多糖（LPS）和持续高氧制备新型BPD模型。选取未作任何处理的正常新生幼鼠18只作为空气对照组，然后将90只新生BPD模型幼鼠分为生理盐水对照组（A 组）、rhEPO800组（B组）、rhEPO1000组（C组）、rhE‐PO1200组（D组）、rhEPO1400组（E组），各18只，在给药后第1、7、14天从各组抽取6只处死并取下肺组织于液氮保存。采用RT‐PCR及Western blot法检测SD新生幼鼠肺组织中eNOS、Bcl‐2蛋白及mRNA水平的表达情况。结果 A组中Bcl‐2蛋白及mRNA表达水平比空气对照组明显降低，而eNOS的表达水平明显升高；B、C、D、E组与A组相比，随着rhEPO浓度的升高，Bcl‐2蛋白和mRNA的表达水平逐渐升高，而 eNOS的表达量逐渐降低，E组中的表达变化最为明显，且差异有统计学意义（ P＜0．05）。结论 rhEPO对新型BPD具有一定的治疗作用，而且连续使用1400 IU／kg rhEPO 14 d ，治疗效果最好。%Objective To observe changes of the expression of Bcl‐2 and endothelial nitric oxide synthase (eNOS) in lung tissue ,which was used the indifferent treatment doses of recombinant human erythropoietin (rhEPO) of newborn SD model of a new type of bronchial pulmonary dysplasia .To explore its effect on the protection of the new type of bronchial pulmonary dysplasia (BPD) ,and find out the appropriate drug delivery time and dosage .Methods We used the things lipopolysaccharide (LPS) and continuous high oxygen preparation to make the new model of the BPD .We made random normal newborn mice 18 as normal air group ,and then 90 newborn baby mice model of random points normal saline group (group A) ,rhEPO800 group (group B);rhEPO 1000 group (group C) ,rhEPO1200 group (group D) ,rhEPO1400 group (group E) ,in experiment 1 ,7 ,and 14 days ,we were ran‐domly selected 6 executed only lung tissue under liquid nitrogen .Using RT‐PCR and Western blot method to detect the Bcl‐2 and eNOS protein and mRNA expression in lung tissue .Results Compared with the normal air group ,the expression of protein and mRNA of Bcl‐2 in group A was decreased ,while the expression of eNOS was increased .Compared with group B ,C ,D ,E ,with the increase of rhEPO concentration ,the expression of Bcl‐2 protein and mRNA in group A gradually increased ,while the expression of eNOS gradually decreased ,and the expression of the group E was the most obvious ,and the difference was statistically significant (P<0 .05) .Conclusion It is prompted that rhEPO has some treatment function to the new BPD ,while the dose of 1 400 IU/kg of 14 days have the best function .