氧化应激诱导鼠羊膜腔衰老和无菌性炎症。
Oxidative stress induces senescence and sterile inflammation in murine amniotic cavity.
机构信息
Division of Maternal-Fetal Medicine and Perinatal Research, Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, United States; Department of Pathology, Botucatu Medical School, UNESP - Univ. Estadual Paulista, Botucatu, Sao Paulo, Brazil.
Division of Maternal-Fetal Medicine and Perinatal Research, Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, United States; Department of Neurobiology, Cell, and Anatomy, The University of Texas Medical Branch at Galveston, Galveston, TX, United States.
出版信息
Placenta. 2018 Mar;63:26-31. doi: 10.1016/j.placenta.2018.01.009. Epub 2018 Feb 2.
OBJECTIVE
A physiologic increase of reactive oxygen species (ROS) is observed through pregnancy. ROS-induced damage to major cellular elements, specifically protein peroxidation, can lead to fetal and placental tissue senescence and inflammation often associated with normal parturition. The purpose of this study was to examine the effects of oxidative stress (OS) in inducing changes in proteins, senescence, and sterile inflammation in pregnant mice.
METHODS
CD-1 mice (n = 5/group) on day 14 of gestation were subjected to minilaparotomy and the uterine horn between gestational sacs was injected with the following: saline (control), cigarette smoke extract (CSE) CSE diluted in saline and CSE + SB 203580 (SB) (a p38 mitogen-activated protein kinase (MAPK) inhibitor). Mice were sacrificed on day 18, and amniotic sacs, placentas and amniotic fluid (AF) were collected. Protein damage was evaluated by immunostaining for 3-Nitrotyrosine modified proteins (3-NT). Activation of prosenescence p38MAPK was evaluated by western blot. Senescence features, β-galactosidase (SA-β-Gal) and AF inflammatory cytokines were analyzed by immunostaining and multiplex luminex-based immunoassays, respectively. The data were analyzed by ANOVA and Tukey's test, p < .05 was used for significance.
RESULTS
Amniotic sac from CSE-treated animals showed significant protein peroxidation compared to control as indicated by 3-NT staining. CSE activated p38MAPK phosphorylation in amniotic sac but not in placenta. Membrane p38MAPK activation was reduced after treatment with SB. CSE increased fetal membrane senescence (staining for SA-β-Gal) and increased AF concentrations of all evaluated cytokines. High inflammation correlated with pup loss and a decrease in placental weight. Treatment with p38MAPK inhibitor (SB) minimized damages, senescence and sterile inflammation.
CONCLUSION
OS induction by cigarette smoke extract cause fetal tissue protein damage, p38MAPK activation, senescence and sterile inflammation in the amniotic cavity of mouse. Prevention of p38MAPK activation can be a novel approach to prevention of adverse pregnancy outcomes related to OS induced premature senescence.
目的
怀孕期间会观察到活性氧（ROS）的生理性增加。ROS 对主要细胞成分的损伤，特别是蛋白质过氧化，可导致胎儿和胎盘组织衰老和炎症，这通常与正常分娩有关。本研究旨在探讨氧化应激（OS）诱导怀孕小鼠蛋白质、衰老和无菌性炎症变化的作用。
方法
妊娠第 14 天的 CD-1 小鼠（每组 5 只）接受小剖腹术，在羊膜囊之间的子宫角内注射以下物质：生理盐水（对照）、香烟烟雾提取物（CSE）、CSE 稀释在盐水中和 CSE+SB 203580（SB）（p38 丝裂原活化蛋白激酶（MAPK）抑制剂）。第 18 天处死小鼠，收集羊膜囊、胎盘和羊水（AF）。通过免疫染色检测 3-硝基酪氨酸修饰蛋白（3-NT）评估蛋白质损伤。通过 Western blot 评估衰老相关的 p38MAPK 激活。通过免疫染色和多重 Luminex 免疫分析分别分析衰老特征、β-半乳糖苷酶（SA-β-Gal）和 AF 炎症细胞因子。数据采用 ANOVA 和 Tukey 检验进行分析，p<0.05 为差异有统计学意义。
结果
与对照相比，CSE 处理的动物的羊膜囊显示出明显的蛋白质过氧化，如 3-NT 染色所示。CSE 在羊膜中激活了 p38MAPK 磷酸化，但在胎盘组织中没有激活。用 SB 处理后，细胞膜 p38MAPK 激活减少。CSE 增加了胎儿膜衰老（SA-β-Gal 染色），并增加了所有评估细胞因子的 AF 浓度。高炎症与幼仔丢失和胎盘重量下降相关。用 p38MAPK 抑制剂（SB）治疗可最大程度地减少损伤、衰老和无菌性炎症。
结论
香烟烟雾提取物引起的氧化应激导致小鼠羊膜腔胎儿组织蛋白质损伤、p38MAPK 激活、衰老和无菌性炎症。抑制 p38MAPK 激活可能是预防与 OS 诱导的过早衰老相关的不良妊娠结局的一种新方法。
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