miR-21通过靶向调控PTEN在新生大鼠急性呼吸窘迫综合征模型中作用的研究  

The role of miR-21 on the targeted regulation of PTEN in neonatal rat model of acute respiratory distress syndrome

作　　者：颜林 梅花[1] 张钰恒 安彩艳[1] 新春 霍梦月 王晓丽 张杰[1] Yan Lin;Mei Hua;Zhang Yuheng;An Caiyan;Xin Chun;Huo Mengyue;Wang Xiaoli;Zhang Jie(Department of Neonatology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院新生儿科,呼和浩特010050

出　　处：《中华新生儿科杂志(中英文)》2025年第2期101-108,共8页Chinese Journal of Neonatology

基　　金：内蒙古自治区重点研发和成果转化计划项目(2022YFSH0088);内蒙古自治区卫生健康委医疗卫生科技计划项目(202201291);内蒙古医科大学重点项目(YKD2022ZD015)。

摘　　要：目的探讨miR-21及PTEN在新生儿急性呼吸窘迫综合征(neonatal acute respiratory distress syndrome,NARDS)大鼠模型中的作用机制。方法将90只7日龄新生SD大鼠分为3组,LPS+miR-21抑制剂组10只、LPS组40只、对照组40只,LPS+miR-21抑制剂组、LPS组采用暴露式气管内滴注4 mg/kg脂多糖(lipopolysaccharide,LPS)的方法构建NARDS大鼠模型,前者同时腹腔注射30 mg/kg miR-21抑制剂,对照组滴注等量等渗NaCl溶液。LPS组及对照组于处理6 h、12 h、24 h、48 h后,LPS+miR-21抑制剂组于24 h后观察大鼠,提取肺组织进行湿重/干重(W/D)比值、HE染色病理评分、实时荧光定量聚合酶链反应检测miR-21和PTEN表达量、WB分析PTEN蛋白表达,ELISA测定白细胞介素(interleukin,IL)6、IL-1β、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)水平。结果1.造模后SD大鼠存活率为100%。随实验进展,LPS组大鼠肺水肿程度、肺病理表现及损伤评分均呈现NARDS肺损伤表现,炎性因子表达逐渐升高,造模成功。2.LPS组miR-21表达量在造模后24 h达峰后下降,而PTEN表达量在24 h降至最低后回升,两者在各时间点表达水平的差异均有统计学意义(P<0.05)。与对照组相比,LPS组中miR-21表达量更高,PTEN表达量更低,差异有统计学意义(P<0.05)。在24 h各组,LPS+miR-21抑制剂组miR-21表达较LPS组显著降低,PTEN表达较LPS组显著增加,差异有统计学意义(P<0.05)。3.LPS组肺组织IL-6、IL-1β、TNF-α表达水平随时间先升高,24 h达峰后下降,均高于对照组、LPS+miR-21抑制剂组(P<0.05)。结论1.暴露式气管内滴注LPS(4 mg/kg)可成功建立NARDS大鼠模型。2.NARDS大鼠模型肺组织中,miR-21与PTEN表达呈时序变化且负相关;同时随miR-21升高、PTEN下降,肺损伤加重、炎性因子表达升高,应用miR-21抑制剂后呈反向趋势,推测miR-21反向调控PTEN介导的NARDS发病进程。ObjectiveTo study the roles of miR-21 and PTEN in the rat model of neonatal acute respiratory distress syndrome(NARDS).MethodsA total of ninety 7 d neonatal SD rats were assigned into the LPS group(n=40),lipopolysaccharide(LPS)+miR-21 inhibitor(miR)group(n=10)and the control group(n=40).NARDS model was established with 4 mg/kg LPS instilled intratracheally.The miR group received an additional 30 mg/kg miR-21 inhibitor intraperitoneal injection when LPS was administered.The control group received intratracheal instillation of same amount of isotonic sodium chloride(NaCl)solution.The LPS group and the control group were observed after 6 h,12 h,24 h and 48 h of treatment and the miR group was observed after 24 h of treatment.The lung tissues were harvested for multiple tests:W/D ratio calculation,histopathological evaluation of lung injury score(LIS)using HE staining,qRT-PCR of miR-21 and PTEN mRNA expression,Western blot of PTEN protein expression and ELISA of IL-6,IL-1βand TNF-αlevels.SPSS 22.0 was used for statistical analysis.ResultsThe survival rate of SD rats was 100%after LPS instillation.The pulmonary edema,pulmonary pathology and LIS,the change of IL-6,TNF-α,IL-1βof the LPS group confirmed successful modeling.In the LPS group,the mRNA of miR-21 decreased after reaching peak at 24 h,while the mRNA of PTEN decreased to the lowest at 24 h and then increased(compared between each timepoint,all P<0.05).Comparing with the control group,the mRNA of miR-21 increased and PTEN decreased in the LPS group(both P<0.05).At 24 h,Comparing with the LPS group,the mRNA of miR-21 significantly decreased and PTEN significantly increased in the miR group(both P<0.05).IL-6,IL-1β,TNF-αin LPS group decreased after reaching peak at 24 h,higher than in the control group and miR group(both P<0.05).ConclusionsIntratracheal instillation of LPS can successfully establish NARDS model in neonatal rats.In the NARDS model,the expression of miR-21 increased and PTEN decreased,sequentially.Additionally,with the increase of miR-21 and th

关 键 词：新生儿 急性呼吸窘迫综合征 MICRORNA-21 PTEN 新生大鼠 

分 类 号：R722.1[医药卫生—儿科]