基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 1024-1029.

• 研究论文 • 上一篇    下一篇

上调蛋白激酶D2表达减轻小鼠肺缺血/再灌注损伤

刘浩明1,华毛1,张莉1,朱海宏2   

  1. 1. 青海省第五人民医院
    2. 青海省人民医院
  • 收稿日期:2020-05-12 修回日期:2020-11-04 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 刘浩明 E-mail:lip56321@163.com
  • 基金资助:
    2015年卫生计生指导性科研课题

Up-regulation gene expression of protein kinase D2 reduces lung ischemia-reperfusion injury in mice

  • Received:2020-05-12 Revised:2020-11-04 Online:2021-07-05 Published:2021-06-17

摘要: 目的 观察蛋白激酶D2(PKD2)表达上调对肺缺血再灌注损伤(LIRI)的改善作用。方法 C57BL/6J野生型(WT)小鼠20只和PKD2基因过表达(PKD2+/+)小鼠40只,随机分为假手术组(WS和PS组)、LIRI组(WIR和PIR组,夹闭左侧肺门1 h,再灌注2 h)及低氧诱导因子-1α(HIF-1α)抑制剂(2-甲氧基雌二醇,2-ME)组(WIR+2ME和PIR+2ME组,造模前2 d腹腔注射20 mg/kg?d 2ME)。再灌注2 h后检测肺组织湿/干重比值(W/D),HE染色观察肺组织病变,ELISA法检测肺组织炎性因子TNF-α、IL-1β和IL-6含量,RT-qPCR和Western blot检测肺组织HIF-1 α和VEGFA mRNA及蛋白表达。结果 与WS组比较,WIR组W/D值增加,肺组织损伤明显,炎性因子含量增加,HIF-1 α和VEGFA mRNA和蛋白表达增加(P<0.05);与WIR组比较,PIR组W/D值降低,肺组织损伤减轻,炎性因子含量降低,HIF-1 α和VEGFA mRNA及蛋白表达增加(P<0.05);与WIR和PIR组比较,WIR+2ME和PIR+2ME组W/D值增加,肺组织损伤加重,炎性因子含量增加,HIF-1 α和VEGFA mRNA及蛋白表达降低(P<0.05),PIR+2ME组上述指标变化与WIR+2ME组无差异。结论 上调PKD2基因可改善LIRI,其机制可能与增加HIF-1 α/VEGFA信号通路的表达有关。

Abstract: Objective To observe the effect of up-regulation of protein kinase D2 (PKD2) expression on lung ischemia-reperfusion injury (LIRI). Methods 20 C57BL/6J wild-type (WT) mice and 40 PKD2 gene overexpression (PKD2+/+) mice were randomly divided into sham operation group (WS and PS groups), LIRI group (WIR and PIR groups, the left hilum was first clamped for 1 h, then reperfused for 2 h) and the hypoxia inducible factor-1α (HIF-1α) inhibitor (2-methoxyestradiol, 2-ME) group (WIR+2ME and PIR+2ME groups, 20 mg/kg?d 2ME was injected intraperitoneally 2 days before modeling). After 2 hours of reperfusion, the wet/dry weight ratio (W/D) of lung tissues was detected, the pathological changes of lung tissues were observed by HE staining, and the content of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissues were detected by ELISA. RT-qPCR and Western blot were used to detect the expression of HIF-1α and VEGFA mRNA and protein in lung tissues. Results Compared with WS group, the W/D value of the WIR group increased, the lung tissues were damaged, the content of inflammatory factors increased, and the expression of HIF-1α and VEGFA mRNA and protein increased (P<0.05). Compared with the WIR group, the PIR group W/D value decreased, lung tissue damage was reduced, inflammatory factor content decreased, HIF-1α and VEGFA mRNA and protein expression increased (P<0.05). Compared with WIR and PIR groups, W/D values of WIR+2ME and PIR+2ME groups increased, lung tissue damage worsened, inflammatory factor content increased, HIF-1α and VEGFA mRNA and protein expression decreased (P<0.05). Compared with WIR+2ME group, there was no significant difference in the changes of the above indicators in PIR+2ME group (P>0.05). Conclusions Up-regulation of PKD2 gene can improve LIRI, and its mechanism may be related to increasing the expression of HIF-1α/VEGFA signaling pathway.