DataSheet_1_A comparison of rat models that best mimic immune-driven preeclampsia in humans.pdf

Preeclampsia (PE), a hypertensive pregnancy disorder, can originate from varied etiology. Placenta malperfusion has long been considered the primary cause of PE. However, we and others have showed that this disorder can also result from heightened inflammation at the maternal-fetal interface. To advance our understanding of this understudied PE subtype, it is important to establish validated rodent models to study the pathophysiology and test therapies. We evaluated three previously described approaches to induce inflammation-mediated PE-like features in pregnant rats: 1) Tumor necrosis factor-α (TNF-α) infusion via osmotic pump from gestational day (GD) 14-19 at 50ng/day/animal; 2) Polyinosinic:polycytidylic acid (Poly I:C) intraperitoneal (IP) injections from GD 10-18 (alternate days) at 10mg/kg/day/animal; and, 3) Lipopolysaccharide (LPS) IP injections from GD 13-18 at 20ug-70ug/kg/day per animal. Maternal blood pressure was measured by tail-cuff. Upon sacrifice, fetal and placenta weights were recorded. Placenta histomorphology was assessed using H&E sections. Placenta inflammation was determined by quantifying TNF-α levels and inflammatory gene expression. Placenta metabolic and mitochondrial health were determined by measuring mitochondrial respiration rates and placenta NAD+/NADH content. Of the three rodent models tested, we found that Poly I:C and LPS decreased both fetal weight and survival; and correlated with a reduction in region specific placenta growth. As the least effective model characterized, TNF-α treatment resulted in a subtle decrease in fetal/placenta weight and placenta mitochondrial respiration. Only the LPS model was able to induce maternal hypertension and exhibited pronounced placenta metabolic and mitochondrial dysfunction, common features of PE. Thus, the rat LPS model was most effective for recapitulating features observed in cases of human inflammatory PE. Future mechanistic and/or therapeutic intervention studies focuses on this distinct PE patient population may benefit from the employment of this rodent model of PE.

子痫前期（Preeclampsia, PE）是一类妊娠高血压疾病，其病因具有多样性。长期以来，胎盘灌注不良被认为是PE的主要致病原因。然而，本团队及其他研究均证实，该疾病亦可由母胎界面炎症反应增强所引发。为加深对这一尚未被充分研究的PE亚型的认识，构建经过验证的啮齿类动物模型，以探究其病理生理学机制并筛选治疗方案，具有重要意义。本研究评估了三种已报道的、可在妊娠大鼠中诱导炎症介导的PE样表型的造模方案：1）自妊娠第14~19天（GD14-19）通过渗透泵持续输注肿瘤坏死因子-α（Tumor necrosis factor-α, TNF-α），剂量为50ng/天/只；2）自GD10~18天每隔一日腹腔内（intraperitoneal, IP）注射聚肌胞苷酸：聚胞苷酸（Polyinosinic:polycytidylic acid, Poly I:C），剂量为10mg/kg/天/只；3）自GD13~18天腹腔内（intraperitoneal, IP）注射脂多糖（Lipopolysaccharide, LPS），剂量为20μg~70μg/kg/天/只。采用尾袖法检测孕鼠血压；处死动物后记录胎鼠与胎盘重量；通过苏木精-伊红（H&E）切片评估胎盘组织形态学特征；通过定量检测TNF-α水平及炎症相关基因表达量，评估胎盘炎症状态；通过测定线粒体呼吸速率及胎盘组织烟酰胺腺嘌呤二核苷酸/还原型烟酰胺腺嘌呤二核苷酸（NAD+/NADH）含量，分析胎盘代谢与线粒体健康状况。在三种受试啮齿类动物模型中，Poly I:C与LPS造模均可降低胎鼠体重与存活率，并与胎盘区域特异性生长受限相关。而TNF-α造模效果最弱，仅可轻微降低胎鼠/胎盘重量及胎盘线粒体呼吸速率。仅LPS造模可诱导孕鼠出现高血压，并表现出显著的胎盘代谢与线粒体功能障碍——这正是PE患者的典型病理特征。因此，大鼠LPS造模模型最能复现人类炎症性PE的临床特征。未来针对这一特殊PE患者群体的机制研究或治疗干预试验，采用该啮齿类PE模型或将获益良多。