PTEN抑制剂BPV通过下调细胞焦亡水平改善LPS诱导的急性肺损伤  

PTEN inhibitor BPV ameliorates LPS induced acute lung injury by inhibiting cell pyroptosis

作　　者：李瑞晟[1] 王琴[1] 李晶晶 穆清爽(指导)[1] LI Ruisheng;WANG Qin;LI Jingjing;MU Qingshuang(Cadre Section 1,the Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830063,China)

机构地区：[1]新疆医科大学第二附属医院干部一科,乌鲁木齐830063

出　　处：《中国免疫学杂志》2023年第5期911-916,共6页Chinese Journal of Immunology

基　　金：新疆维吾尔自治区自然科学基金项目(2021D01C371)。

摘　　要：目的:探究第10号染色体丢失的张力蛋白同源磷酸酶基因(PTEN)抑制剂过氧化氢氧化矾酸钾(BPV)对脂多糖(LPS)诱导的急性肺损伤(ALI)的影响及作用机制。方法:将60只雄性C57BL/6小鼠按随机数字表法分为假手术组(Sham组)、BPV组、LPS组和BPV+LPS组,其中又将LPS组分为LPS刺激后6 h、12 h、24 h、48 h组;Western blot检测LPS各组小鼠肺组织中PTEN蛋白表达变化;检测小鼠肺组织湿/干重(W/D),分析Sham组,BPV组、LPS组及BPV+LPS组小鼠肺组织水肿情况;HE染色观察4组小鼠肺组织病理学改变;ELISA和流式细胞术分别检测4组小鼠肺组织灌洗液(BALF)中炎症因子IL-1β、IL-6、IL-18和TNF-α的表达水平及巨噬细胞数量变化;组织免疫荧光染色检测各组小鼠肺泡巨噬细胞中pyrin结构域NLRP3表达;Western blot检测4组小鼠肺组织中NLRP3介导的焦亡途径中NLRP3、caspase-1 p10、ASC和GSDMD p30蛋白表达及PTEN/Akt/NF-κB信号通路中PTEN蛋白、Akt^(s473)的磷酸化水平及TLR4和NF-κB p65的蛋白表达。结果:与Sham组相比,LPS能以时间依赖性刺激小鼠肺组织中PTEN表达,且作用24 h的促进效果最为显著(P<0.01),并诱导小鼠肺组织损伤(P<0.05),促进BALF中巨噬细胞比例及炎症因子IL-1β、IL-6、IL-18和TNF-α表达(P<0.05或P<0.01),同时肺组织巨噬细胞中NLRP3表达明显增加(P<0.05);与LPS组相比,BPV+LPS组小鼠肺组织损伤明显减轻(P<0.05),BALF中上述炎症因子表达水平显著降低(P<0.05),肺泡巨噬细胞中NLRP3表达亦明显降低(P<0.05)。Western blot结果显示,与Sham组相比,LPS组和LPS+BPV组中NL-RP3、caspase-1 p10、ASC和GSDMD p30、PTEN蛋白及Akt^(s473)的磷酸化水平及TLR4和NF-κB p65表达均显著升高(P<0.05或P<0.01);与LPS组相比,除Akt^(s473)的磷酸化水平明显升高外(P<0.05),其他蛋白水平均明显降低(P<0.05)。结论:PTEN抑制剂BPV可能通过Akt/NF-κB信号通路抑制肺组织中NLRP3介导的细胞焦亡,改善LPS诱导的小鼠ALI症状�Objective:To explore effect and mechanism of phosphatase and tensin homology deleted on chromosome ten(PTEN)inhibitor potassium bisperoxo(1,10-phenanthroline)oxovanadate(BPV)on lipopolysaccharide(LPS)induced acute lung injury(ALI).Methods:Sixty male C57BL/6 mice were randomly divided into sham operation group(Sham group),BPV group,LPS group and BPV+LPS group,the LPS group was further divided into 6 h,12 h,24 h and 48 h groups after LPS stimulation;Western blot was used to detect the expression of PTEN protein in lung tissue of mice in LPS groups;Wet dry ratio(W/D)of lung tissue was detected to analysis edema of lung tissues in Sham group,BPV group,LPS group and BPV+LPS group;HE staining was used to observe pathological changes of lung tissue in four groups;expression levels of IL-1β,IL-6,IL-18 and TNF-αin BALF and the number of macrophages were detected by ELISA and flow cytometry;expression of pyrin domain NLRP3 in alveolar macrophages was detected by tissue immunofluorescence staining;protein expressions of NLRP3,caspase-1 p10,ASC and GSDMD p30,PTEN protein,phosphorylation level of Akt^(s473) and TLR4 and NF-κB p65 protein in PTEN/Akt/NF-κB signaling pathway were detected by Western blot.Results:Compared with Sham group,LPS could stimulate expression of PTEN in lung tissue of mice in a time-dependent manner,the effect was the most significant at 24 h(P<0.01),it also induced lung tissue injury in mice(P<0.05),increased proportion of macrophages in BALF and expressions of inflammatory factors IL-1β,IL-6,IL-18 and TNF-α(P<0.05,P<0.01);at the same time,expression of NLRP3 in alveolar macrophages was increased significantly(P<0.05).Compared with LPS group,lung injury of BPV+LPS group was significantly reduced(P<0.05),expressions of above inflammatory factors in BALF were significantly decreased(P<0.05),and expression of NLRP3 in alveolar macrophages was also significantly decreased(P<0.05).Western blot results showed that compared with Sham group, expression levels of NLRP3, caspase-1 p10, ASC and GSDMD p30, P

关 键 词：第10号染色体丢失的张力蛋白同源磷酸酶基因抑制剂 急性肺损伤 巨噬细胞 焦亡 

分 类 号：R563[医药卫生—呼吸系统]