上调蛋白激酶D2表达减轻小鼠肺缺血/再灌注损伤  

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

作　　者：刘浩明 华毛 张莉 朱海宏[3] LIU Hao-ming;HUA Mao;ZHANG Li;ZHU Hai-hong(Department of Thoracic Surgery,the Fifth People's Hospital of Qinghai Province,Xining 810007;Head and Neck Surgery, the Fifth People's Hospital of Qinghai Province, Xining 810007;Department of Thoracic Surgery, Qinghai People's Hospital, Xining 810007, China)

机构地区：[1]青海省第五人民医院胸外科,青海西宁810007 [2]青海省第五人民医院头颈外科,青海西宁810007 [3]青海省人民医院胸外科,青海西宁810007

出　　处：《基础医学与临床》2021年第7期1024-1029,共6页Basic and Clinical Medicine

基　　金：2015年卫生计生指导性科研课题基金([2015]18-17)。

摘　　要：目的观察蛋白激酶D2(PKD2)表达上调对小鼠肺缺血/再灌注损伤(LI/RI)的改善作用。方法C57BL/6J野生型(WT)小鼠20只和PKD2过表达(PKD2+/+)小鼠40只,随机分为假手术组(WS和PS组)、LI/RI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[W(IR+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(W/D)值,HE染色观察肺组织病变,ELISA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-1α、血管内皮生长因子A(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-1α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-1α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-1α和VEGFA mRNA及蛋白表达降低(P<0.05)。结论上调PKD2基因可改善LI/RI,其机制可能与增加HIF-1α/VEGFA信号通路的表达有关。Objective To observe the effect of up-regulation of protein kinase D2(PKD2)expression on lung ischemia-reperfusion injury(LI/RI)in mice.Methods 20 C57BL/6J wild-type(WT)mice and 40 PKD2 over-expression(PKD2+/+)mice were randomly divided into sham operation group(WS and PS groups),LI/RI 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-1βand IL-6 in lung tissues were detected by ELISA.RT-qPCRand Western blot were used to detect the expression of(HIF-1α)and vascular endothelial growth factor A(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-1αand 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-1αand 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-1αand VEGFA mRNA and protein expression significantly decreased(P<0.05).Conclusions Up-regulation of PKD2 expression can improve LI/RI,and its mechanism may be related to increasing the expression of HIF-1α/VEGFA signaling pathway.

关 键 词：肺缺血/再灌注损伤 蛋白激酶D2 HIF-1α/VEGFA信号通路 2-甲氧基雌二醇 

分 类 号：R363[医药卫生—病理学]