hAMSCs调控MAPK信号通路对急性肺损伤AQP1的影响  

Effect of human amniotic mesenchymal stem cell regulating MAPK signaling pathway of aquaporin 1 in acute lung injury

作　　者：李埝 赵建军 张建勇 赵睿桢 Li nian;Zhao Jianjun;Zhang Jianyong;Zhao Ruizhen(The Second Ward,Department of Respiratory and Critical Care Medicine,Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学附属医院呼吸与危重症医学科呼吸二病区,遵义563000

出　　处：《中华肺部疾病杂志（电子版）》2023年第2期156-163,共8页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：黔科合支撑[2020]4Y162号。

摘　　要：目的 分析人羊膜间充质干细胞(human amniotic mesenchymal stromal cells, hAMSCs)移植后对急性肺损伤(acute lung injury, ALI)大鼠肺组织水通道蛋白1(AQP1)和磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)表达的影响及对肺损伤的作用机制。方法 随机将84只SD雄性大鼠分为4组(n=21):生理盐水对照组(NS组)、LPS诱导ALI模型组(LPS组)、hAMSCs对照组(NH组)及hAMSCs干预组(LH组)。LPS组和LH组大鼠通过舌下静脉注射LPS(4 mg/kg)造模;NS组和NH组大鼠通过舌下静脉注射250μl生理盐水,2 h后NH组和LH组大鼠通过尾静脉注射250μl经DAPI标记的hAMSCs(2.5×10^(6)个);NS组和LPS组大鼠通过尾静脉注射250μl生理盐水。每组分别于6 h、12 h、72 h随机处死7只大鼠观察hAMSCs在每组中的定植情况、肺组织病理形态,湿干重比值(W/D),Elisa法检测BALF中TNF-α、IL-1β水平,IHC及WB法检测各组大鼠肺组织AQP1、p38MAPK和p-p38 MAPK的表达。结果 (1)NH和LH组大鼠肺泡周围有蓝色荧光标记的hAMSCs, LH组分布较多,NH组分布较少;(2)NS组、NH组对应时相点的大鼠肺组织病理评分、W/D比值、BALF中TNF-α和IL-1β水平、AQP1和p-p38 MAPK蛋白表达量相比较无显著性差异(P>0.05);(3)与NS组、NH组时相点相比较,LPS组、LH组对应时相点大鼠肺组织病理评分、W/D比值、BALF中TNF-α、IL-1β水平和p-p38 MAPK蛋白表达量升高,AQP1表达量降低(P<0.05);(4)LPS 6 h组与12 h组大鼠肺组织病理评分、W/D比值、BALF中TNF-α和IL-1β水平、AQP1和p-p38 MAPK蛋白表达量无明显差异(P>0.05);与LPS 6 h组、12 h组相比较,LPS 72 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水平、p-p38 MAPK蛋白表达量降低,AQP1表达量升高(P<0.05);LH 6 h组与12 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水平、AQP1和p-p38 MAPK蛋白表达量无明显差异(P>0.05);与LH 6 h组、12 h组相比较,LH 72 h组大鼠肺组织病理评分、W/D比值、BALF中IL-1β和TNF-α水Objective To investigate the effect of the transplantation of human amniotic mesenchymal stem cell(hAMSCs)on the expressions of Aquaporin 1(AQP1)and phosphorylated p38 mitogen activated protein kinase(p-p38MAPK)in lung tissue of rats with acute lung injury(ALI),and to further explore whether it can alleviate lung injury and its possible mechanism.Methods 84 male SD rats were divided into 4 groups(n=21):normal saline control group(NS group),LPS induced ALI model group(LPS group),hAMSCs control group(NH group)and hAMSCs intervention group(LH group).The rats in LPS group and LH group were injected with LPS(4 mg/kg)through a sublingual vein to create models.NS group and NH group were injected with 250μl normal saline through a sublingual vein,and two hours later NH group and LH group were injected with 250μl DAP-labeled hAMSCs(2.5×10^(6))through a caudal vein.NS group and LPS group were injected with 250μl normal saline through a caudal vein.Seven rats in each group were killed at 6 h,12 h and 72 h in order to observe the colonization of hAMSCs in the lung tissue of rats,the pathological morphology of lung tissue and the lung wet/dry weight(W/D)ratio.Furthermore,the levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in bronchoalveolar lavage fluid(BALF)of rats in each group were detected by enzyme linked immunosorbent assay(ELISA).Immunohistochemistry(IHC)and western blot(WB)was used to detect the expressions of AQP1,p38 MAPK and p-p38 MAPK in the lung tissue of each group.Results①Under the fluorescence microscope,it could be observed that there were blue fluorescent labeled hAMSCs around the alveoli of rats in NH and LH groups.Among them,the distribution of hAMSCs in LH group is more than that in NH group,indicating that hAMSCs can"homing"and colonize in the injured lung tissue.②There was no significant difference in lung histopathological score,W/D ratio,TNF-αand IL-1βlevels in BALF,AQP1 and p-p38 MAPK protein expressions between NS group and NH group at each time point(P>0.05).③Compar

关 键 词：人羊膜间充质干细胞 脂多糖 急性肺损伤 水通道蛋白1 

分 类 号：R563[医药卫生—呼吸系统]