微小RNA-21对脓毒症急性肺损伤小鼠肺组织的修复作用及其与PTEN的关系  

Protective effect of microRNA-21 on acute lung injury mice induced by sepsis and its relationship with PTEN

作　　者：葛晨 刘俊行 付优[1] 贾丽静 龙玲[1] 杜全胜[1] GE Chen;LIU Junhang;FU You;JIA Lijing;LONG Ling;DU Quansheng(Department of Intensive Care Unit,Hebei General Hospital,Shijiazhuang 050000,China;不详)

机构地区：[1]河北省人民医院重症医学科,石家庄050000 [2]河北省儿童医院骨科

出　　处：《山东医药》2022年第2期30-34,共5页Shandong Medical Journal

基　　金：河北省医学科学研究重点课题(20180192)。

摘　　要：目的观察脓毒症急性肺损伤(ALI)小鼠肺组织中微小RNA-21(miR-21)的表达情况,以及干预miR-21表达对肺水肿、血清炎症因子及氧化应激的影响,探讨该作用机制与PTEN的关系。方法取96只健康雄性昆明小鼠,采用随机数字表法分为6组:假手术组(n=32)、模型组(n=32)、miR-21前体组(MP组,n=8)、miR-21抑制剂组(MI组,n=8)、miR-21前体+PTEN抑制剂组(MP+Anti-PTEN组,n=8)、miR-21抑制剂+PTEN抑制剂组(MI+Anti-PTEN组,n=8)。假手术组仅进行开腹探查盲肠;模型组采用盲肠结扎穿孔术制作ALI模型;MP组、MI组分别在尾静脉注射miR-21前体腺病毒、miR-21抑制剂腺病毒4 d后再造模;MP+Anti-PTEN组、MI+Anti-PTEN组分别在转染miR-21前体、miR-21抑制剂后,造模前30 min给予尾静脉注射PTEN抑制剂。造模后24 h,取小鼠腹主动脉取血进行血气分析,计算氧合指数(OI)。取小鼠左肺组织,称取肺湿重和肺干重,计算肺湿重/干重比值(W/D)。取小鼠腹主动脉血,采用ELISA法检测血清炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和氧化应激指标活性氧(ROS)、超氧化物歧化酶(SOD)水平。分别于造模后6、12、24、48 h采用qRT-PCR法检测假手术组、模型组肺组织miR-21的相对表达量。于造模后24 h对假手术组、模型组、MP组、MI组采用qRT-PCR法检测肺组织miR-21表达,Western blotting法检测肺组织PTEN蛋白表达。结果造模后24 h,与假手术组比较,模型组OI下降,W/D、TNF-α、IL-6、ROS、SOD水平升高(P均<0.05);与模型组比较,MP组、MP+Anti-PTEN组、MI+AntiPTEN组OI升高,W/D、TNF-α、IL-6、ROS、SOD水平下降(P均<0.05),MI组OI下降,W/D、TNF-α、IL-6、ROS、SOD水平升高(P均<0.05);与MP+Anti-PTEN组比较,MI+Anti-PTEN组OI、W/D、TNF-α、IL-6、ROS、SOD水平无明显变化(P均>0.05)。与假手术组比较,模型组造模后6、12、24、48 h肺组织miR-21表达均升高(P均<0.05),并于造模后24 h达峰。造模后24 h,与假手术组比较,模�Objective To observe the expression of microRNA-21(miR-21) in acute lung injury(ALI) mice induced by sepsis,to explore the effects of intervention of miR-21 expression on pulmonary edema,serum inflammatory factors and oxidative stress,and to investigate their relationship with phosphatase and tensin homolog(PTEN).Methods Ninety-six healthy male Kunming mice were randomly divided into six groups: sham operation group(Sham group,n =32),sepsis-induced ALI group(ALI group,n = 32),miR-21 precursor group(MP group,n = 8),miR-21 inhibition group(MI group,n = 8),miR-21 precursor + PTEN inhibition group(MP + Anti-PTEN group,n = 8),and miR-21 inhibition + PTEN inhibition group(MI + Anti-PTEN group,n = 8).In the Sham group,laparotomy was performed to explore thececum.Mice in the ALI group were induced by cecal ligation puncture(CLP).Mice in the MP group and MI group were first injected with miR-21 precursor adenovirus and miR-21 inhibitor adenovirus,respectively,and after 4 d,models were established.Mice in the MP + Anti-PTEN group and MI + Anti-PTEN group were given PTEN inhibitor by caudal vein 30 minutes before CLP on the basis of transfection of miR-21 precursor and miR-21 inhibitor.The blood samples were collected for determination of oxygenation index(OI),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),reactive oxygen specie(ROS),and superoxide dismutase(SOD) levels by ELISA,and the lung samples for determination of wet to dry weight ratio(W/D) at 24 h in each group.Mice were sacrificed and lungs were removed for determination of miR-21 expression(by qRT-PCR) in the Sham and ALI group at 6,12,24 and 48 h.Lung tissues were collected for determination of miR-21 expression(by qRT-PCR) and PTEN expression(by Western blotting) in the Sham,ALI,MP and MI groups at24 h.Results Compared with the Sham group,OI decreased,W/D,TNF-α,IL-6,ROS and SOD levels increased in the ALI group at 24 h after modeling(all P < 0.05).Compared with the ALI group,OI increased,W/D,TNF-α,IL-6,ROS,and SOD levels decreased in the MP,MP + Anti-PT

关 键 词：脓毒症 急性肺损伤 微小RNA-21 PTEN蛋白 炎症因子 氧化应激 小鼠 

分 类 号：R631[医药卫生—外科学]