目的观察百草枯(PQ)中毒导致的急性肺损伤后内皮祖细胞(EPCs)移植治疗对大鼠肺组织丝裂原活化蛋白激酶(MAPK)通路蛋白表达的影响。方法36只SD雄性大鼠按随机数字表法平均分成三组:正常对照组、PQ损伤组和EPCs治疗组,每组12只。EPCs移植后24 h处死大鼠并收集大鼠肺组织和血清。分别采用比色法测定肺组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平,酶联免疫吸附(ELISA)法检测血清中细胞炎性因子白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量,血气分析仪检测大鼠动脉血氧分压(PaO2),肺组织切片后通过HE染色对肺损伤程度进行评估,Western blot方法测定肺组织中MAPK信号通路蛋白表达。结果与对照组比较,PQ损伤组大鼠血清IL-1β(μg/L:25.16±1.71 vs.34.94±1.68)和TNF-α(μg/L:51.17±2.33 vs.78.46±5.16)含量明显增加(P<0.05),PQ损伤组大鼠肺组织中MDA(nmol/m L:3.08±0.19 vs.6.89±0.44)含量明显增加(P<0.05),但是SOD(U/m L:249.11±4.71 vs.191.15±3.21)活性降低,且PaO2(mm Hg:92.19±3.31 vs.40.18±5.54)下降(P<0.05);与PQ损伤组比较,EPCs治疗组大鼠SOD(U/m L:191.15±3.21 vs.216.64±2.39)活性升高(P<0.05),MDA(nmol/m L:6.89±0.44 vs.5.13±0.40)、IL-1β(μg/L:34.94±1.68 vs.26.11±1.43)和TNF-α(μg/L:78.46±5.16 vs.60.18±4.31)含量降低(P<0.05),PaO2(mm Hg:40.18±5.54 vs.56.81±5.16)升高(P<0.05)。MAPK通路蛋白的磷酸化水平在对照组中仅有少量表达,而给予百草枯后表达量显著升高(P<0.05),使用EPCs治疗后各蛋白表达量又显著降低(P<0.05)。结论百草枯中毒可激活包含p38MAPK、C-JUN氨基末端激酶(JNK)、细胞外调节蛋白激酶1/2(ERK1/2)在内的MAPK通路。EPCs可以改善PQ中毒大鼠症状,减少肺部损伤,改善供氧,其机制可能与抑制MAPK通路各蛋白磷酸化有关。Objective To observe the effect of endothelial progenitor cells(EPCs) transplantation after acute lung injury caused by paraquat(PQ)poisoning on the protein expression of mitogen-activated protein kinase(MAPK)pathway protein in rat lung tissue.Methods 36 SD male rats were randomly divided into 3 groups according to the table of random numbers:normal control group(n=12),PQ injury group(n=12)and EPCs treatment group(n=12).The rats were killed 24 hours after EPCs transplantation,lung tissue and serum were collected.Colorimetric method was used to determine the superoxide dismutase(SOD)activity and malondialdehyde(MDA)level in lung tissue,enzyme-linked immunoassay to detect cell inflammatory cytokines of serum interleukin-1 beta(IL-1β)and tumor necrosis factor-alpha(TNF-α),blood gas analyzer to test rat arterial partial pressure of oxygen(PaO2).The degree of lung injury was evaluated by lung tissue section after HE staining.The protein expression of MAPK signaling pathway in lung tissue was determined by Western blot.Results After PQ poisoning,compared with control group,serum IL-1β(μg/L:25.16±1.71 vs.34.94±1.68)and TNF-α(μg/L:51.17±2.33 vs.78.46±5.16)levels of rats in PQ injury group increased significantly(P<0.05).MDA of rat lung tissue in PQ injury group(nmol/m L:3.08±0.19 vs.6.89±0.44)increased significantly(P<0.05),but the activity of SOD(U/m L:249.11±4.71 vs.191.15±3.21)and PaO2(mm Hg:92.19±3.31 vs.40.18±5.54)decreased(P<0.05).After EPCs treatment,compared with PQ injury group,the activity SOD(U/m L:191.15±3.21 vs.216.64±2.39)in EPCs treatment group increased(P<0.05),the content of MDA(nmol/m L:6.89±0.44 vs.5.13±0.40),IL-1β(μg/L:34.94±1.68 vs.26.11±1.43)and TNF-α(μg/L:78.46±5.16 vs.60.18±4.31)decreased(P<0.05),PaO2(mm Hg:40.18±5.54 vs.56.81±5.16)gradually increased(P<0.05).The phosphorylation level of MAPK pathway protein was only slightly expressed in control group.After giving PQ,the expression of PQ injury group was significantly increased(P<0.05),and the protein expres
机构地区中国医科大学附属第一医院急诊科
出处《中国急救医学》 CAS CSCD 北大核心 2021年第2期161-165,共5页Chinese Journal of Critical Care Medicine
基金国家自然科学基金资助项目(81772053)。
关键词内皮祖细胞(EPCs) 移植 百草枯(PQ) 中毒 急性肺损伤(ALI) 氧化应激 炎性因子 丝裂原活化蛋白激酶通路蛋白Endothelial progenitor cell(EPCs) Transplantation Paraquat(PQ) Poisoning Acute lung injury(ALI) Oxidative stress Inflammatory factors Mitogen-activated protein kinase(MAPK)pathway protein
分类号R28 [医药卫生—中药学]
