PI3K/Akt信号通路在骨髓间充质干细胞条件培养基减轻大鼠内毒素性急性肺损伤中的作用  

Role of PI3K/Akt signaling pathway in reduction of acute lung injury by bone marrow mesenchymal stem cells-conditioned medium in rats

作　　者：罗思静 林群[2] 雷立华[3] Luo Sijing;Lin Qun;Lei Lihua(Department of Anesthesiology,The Affiliated TCM Hospital of Guangzhou Medical University,Guangzhou 510130,China;Department of Anesthesiology,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China;Department of Anesthesiology,Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,China)

机构地区：[1]广州医科大学附属中医医院麻醉科,广州510130 [2]福建医科大学附属第一医院麻醉科,福州350005 [3]福建医科大学省立临床学院麻醉科,福州350001

出　　处：《中华麻醉学杂志》2022年第11期1375-1379,共5页Chinese Journal of Anesthesiology

基　　金：福建省财政厅教育和科研专项资金(闽财指【2021】848号)。

摘　　要：目的:评价磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路在骨髓间充质干细胞(MSCs)条件培养基减轻大鼠内毒素性急性肺损伤中的作用。方法:健康雄性SD大鼠32只,6~8周龄,体重200~250 g,按照随机数字表法分为4组(n=8):对照组(C组)、急性肺损伤组(ALI组)、急性肺损伤+MSCs条件培养基组(M组)和急性肺损伤+MSCs条件培养基+LY294002组(L组)。ALI组、M组和L组采用气管内雾化吸入LPS 5 mg/kg的方法制备大鼠急性肺损伤模型。给予LPS 1 h后,C组和ALI组静脉注射无血清DMEM培养基1 ml;M组静脉注射MSCs条件培养基1 ml;L组于造模前2 h腹腔注射PI3K抑制剂LY29400230 mg/kg,造模后1 h静脉注射MSCs条件培养基1 ml。给予LPS后24 h,心脏取血,分离血清,采用ELISA法测定血清TNF-α和IL-6的浓度;然后处死大鼠,取肺组织,测定湿重/干重(W/D)比值,光镜下观察肺组织病理学结果,并行肺损伤评分,采用Western blot法测定Akt和磷酸化Akt(p-Akt)的表达,计算p-Akt/Akt比值。结果:与C组比较,ALI组肺W/D比值、肺损伤评分、血清TNF-α和IL-6浓度升高,肺组织p-Akt表达下调,p-Akt/Akt比值下降(P<0.05),肺组织发生病理学损伤;与ALI组比较,M组肺W/D比值、肺损伤评分、血清TNF-α和IL-6浓度下降,肺组织p-Akt表达上调,p-Akt/Akt比值升高(P<0.05),肺组织病理学损伤减轻,L组上述各指标差异无统计学意义(P>0.05);与M组比较,L组肺W/D比值、肺损伤评分、血清TNF-α和IL-6浓度升高,肺组织p-Akt表达下调,p-Akt/Akt比值下降(P<0.05),肺组织病理学损伤加重。4组间肺组织Akt表达比较差异无统计学意义(P>0.05)。结论:PI3K/Akt信号通路参与了MSCs条件培养基减轻大鼠内毒素性急性肺损伤的过程。Objective To evaluate the role of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway in reduction of acute lung injury(ALI)by bone marrow mesenchymal stem cells(MSCs)-conditioned medium in rats.Methods Thirty-two healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-250 g,were divided into 4 groups(n=8 each)according to the random number table method:control group(group C),group ALI,ALI+bone marrow MSCs-conditioned medium group(group M),and ALI+bone marrow MSCs-conditioned medium+LY294002 group(group L).A rat model of ALI was developed by endotracheal inhalation of aerosolized lipopolysaccharide(LPS)5 mg/kg in anesthetized animals in group ALI,group M and group L.At 1 h after LPS administration,serum-free DMEM medium 1 ml was intravenously injected in group C and group ALI,bone marrow MSCs-conditioned medium 1 ml was intravenously injected in group M,PI3K inhibitor LY29400230 mg/kg was intraperitoneally injected at 2 h before the model was developed,and bone marrow MSCs-conditioned medium 1 ml was intravenously injected at 1 h after the model was developed in group L.The blood samples were taken from the heart at 24 h after LPS administration,and serum was isolated for determination of the serum levels of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)(by enzyme-linked immunosorbent assay).Then the rats were sacrificed and lung tissues were obtained for determination of the lung wet/dry weight ratio(W/D ratio),expression of Akt and phosphorylated Akt(p-Akt)(by Western blot)and for microscopic examination of the pathological changes which were scored.The p-Akt/Akt ratio was calculated.Results Compared with group C,the W/D ratio,lung injury score and serum levels of TNF-αand IL-6 were significantly increased,the expression of p-Akt in lung tissues was down-regulated,the ratio of p-Akt/Akt was decreased(P<0.05),and pathological changes were found in lung tissues in group ALI.Compared with group ALI,the W/D ratio,lung injury score and serum levels of TNF-αand IL-6 were signi

关 键 词：磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 间质干细胞 骨髓 内毒素类 急性肺损伤 

分 类 号：R563.8[医药卫生—呼吸系统]