下调miR-128-3p缓解脓毒症大鼠急性肺损伤的炎症反应和肺组织形态学的影响  

Alleviation of the inflammatory response induced by acute lung injury in sepsis rats by miR-128-3p down-regulation and effects on lung tissue morphology

作　　者：黄钟[1] 孙洁[2] 姚振滨[1] 李桂花[1] 许晓刚[1] HUANG Zhong;SUN Jie;YAO Zhenbin;LI Guihua;XU Xiaogang(the First Affiliated Hospital of Shihezi University Medical College Department of Emergency Medicine,Shihezi 832000,China;the First Affiliated Hospital of Shihezi University Medical College Nosocomial Infection Control Office,Shihezi 832000)

机构地区：[1]石河子大学医学院第一附属医院急诊内科,新疆石河子832000 [2]石河子大学医学院第一附属医院,院内感染控制办公室,新疆石河子832000

出　　处：《中国比较医学杂志》2020年第12期9-16,共8页Chinese Journal of Comparative Medicine

基　　金：新疆维吾尔自治区地区科学基金项目(81260363)。

摘　　要：目的探究下调miR-128-3p缓解脓毒症大鼠急性肺损伤的炎症反应和肺组织形态学的影响。方法选取60只健康3周龄SD大鼠,采用随机分组法将大鼠分为:假手术组、脓毒症模型组、空白转染组和miR-128-3p沉默组,每组15只。除假手术组外各组大鼠采用盲肠结扎穿孔法制备大鼠脓毒症模型;空白转染组构建空白质粒并转染大鼠;miR-128-3p沉默组构建慢病毒质粒并转染大鼠。RT-PCR检测miR-128-3p表达,确定干扰成功;检测各组大鼠静息通气量、气道阻力改变和肺容积变换情况;采用ELISA检测外周血清IL-6、TNF-α、iNOS的含量;采用HE染色观察肺上皮组织纤维化情况及病理损伤情况;采用TUNEL染色结合蛋白质印记检测肺组织中Caspase-3和Caspase-9的表达;采用MASSON染色结合蛋白质印记检测TGF-β和α-SMA的表达情况。结果HE染色结果显示:miR-128-3p沉默组细胞分布较为均匀,且坏死细胞较少。MASSON染色果显示:脓毒症模型组肺组织出现严重纤维化和损伤,miR-128-3p沉默组肺组织纤维化程度得到改善。相比假手术组,脓毒症模型组大鼠miR-128-3p表达、气道阻力、IL-6含量水平、iNOS含量水平、TNF-α含量水平、肺组织细胞凋亡率、cleaved cas3/Caspase-3、cleaved cas9/Caspase-9、TGF-β蛋白相对表达和α-SMA蛋白相对表达均显著升高(P<0.05),静息通气量、气道阻力改变和肺容积变换显著降低(P<0.05)。相比空白转染组,miR-128-3p沉默组miR-128-3p表达、气道阻力、IL-6含量水平、iNOS含量水平、TNF-α含量水平、肺组织细胞凋亡率、cleaved cas3/Caspase-3、cleaved cas9/Caspase-9、TGF-β蛋白相对表达和α-SMA蛋白相对表达均显著降低(P<0.05),静息通气量、气道阻力改变和肺容积变换显著升高(P<0.05)。结论下调miR-128-3p可以缓解脓毒症大鼠急性肺损伤,其作用机制可能与降低大鼠体内炎症反应并抑制肺组织细胞的凋亡相关。Objective To explore alleviation of the inflammatory response induced by acute lung injury in sepsis rats by down-regulation of miR-128-3p and effects on lung tissue morphology.Methods Sixty healthy 3-week-old SD rats were divided into a sham-operated group,a sepsis model group,a blank transfection group and an miR-128-3p inhibitor group by a random grouping method with 15 rats in each group.Sepsis models were prepared by cecal ligation and puncture.In the blank transfection group,blank plasmids were constructed and were transfected into rats.In the miR-128-3p inhibitor group,lentiviral plasmids were constructed and were transfected into rats.The expression of miR-128-3p was detected by RT-PCR to determine the success of interference.Resting ventilation,changes in airway resistance and lung volume were determined in each group.The contents of serum IL-6,TNF-α and iNOS in peripheral blood were detected by ELISA.Lung tissues and pathological injury were observed by hematoxylin&eosin(HE)staining.The expression of Caspase-3 and Caspase-9 in lung tissue was detected by TUNEL(terminal deoxynucleotidyl transferase dUTP nick end labeling)staining and western blotting.The expression of TGF-β and α-SMA was detected by Masson's trichrome staining and western blotting.Results HE staining showed that cell distribution was even in the miR-128-3p inhibitor group,with few necrotic cells.MASSON staining showed that there was severe fibrosis and damage of lung tissues in sepsis model group,and degree of lung tissue fibrosis in miR-128-3p silencing group was improved.Compared with the sham-operated group,expression of miR-128-3p,airway resistance,levels of IL-6,iNOS and TNF-α,apoptosis rate of lung tissue cells,relative expression of cleared cas3/Caspase-3,cleared cas9/Caspase-9,and TGF-β and α-SMA protein levels were significantly increased(P<0.05),while resting ventilation,changes in airway resistance and lung volume were significantly decreased in the sepsis model group(P<0.05).Compared with the blank transfection group,ex

关 键 词：脓毒症 急性肺损伤 组织形态学 炎症反应 

分 类 号：R-33[医药卫生]