Kadam Leena, Gomez-Lopez Nardhy, Mial Tara N, Kohan-Ghadr Hamid-Reza, Drewlo Sascha
1 Department of Physiology, School of Medicine, Wayne State University, Detroit, MI, USA.
2 Department of Obstetrics and Gynecology, School of Medicine, Wayne State University, Detroit, MI, USA.
Reprod Sci. 2017 Dec;24(12):1590-1599. doi: 10.1177/1933719117697128. Epub 2017 Mar 21.
Elevated inflammation accounts for approximately 30% of preterm birth (PTB) cases. We previously reported that targeting the peroxisome proliferator-activated receptor gamma (PPARγ) pathway reduced the incidence of PTB in the mouse model of endotoxin-induced PTB. The PPARγ has proven anti-inflammatory functions and its activation via rosiglitazone significantly downregulated the systemic inflammatory response and reduced PTB and stillbirth rate by 30% and 41%, respectively, in our model. Oxidative stress is inseparable from inflammation, and rosiglitazone has a reported antioxidative activity. In the current study, we therefore aimed to evaluate whether rosiglitazone treatment had effects outside of inflammatory pathway, specifically on the antioxidation pathway in our model.
Pregnant C57BL/6J mice (E16.5) were treated with phosphate-buffered saline (PBS), rosiglitazone (Rosi), lipopolysaccharide (LPS; 10µg in 200µL 1XPBS), or LPS + Rosi (6 hours after the LPS injection). The myometrial and decidual tissues were collected and processed for macrophage isolation using magnetic cell sorting and F4/80+ antibody. Expression levels of antioxidative factors- Nrf2 and Ho-1-along with the LPS receptor Tlr4 were quantified by quantitative polymerase chain reaction. The protein levels were assessed by immunofluorescence staining.
Both the decidual and myometrial macrophages from the LPS-treated animals showed significantly lowered expression of Ho-1 and Nrf2 and higher expression of Tlr4 when compared to the PBS control group. The macrophages from the animals in the LPS + Rosi group had significantly elevated expression of Ho-1 and Nrf2 and downregulated expression of Tlr4 when compared to the LPS group.
Rosiglitazone administration prevents PTB by downregulating inflammation and upregulating antioxidative response.
炎症升高约占早产(PTB)病例的30%。我们之前报道,靶向过氧化物酶体增殖物激活受体γ(PPARγ)途径可降低内毒素诱导的PTB小鼠模型中的PTB发生率。PPARγ已被证明具有抗炎功能,在我们的模型中,通过罗格列酮激活PPARγ可显著下调全身炎症反应,并使PTB和死产率分别降低30%和41%。氧化应激与炎症密不可分,且有报道称罗格列酮具有抗氧化活性。因此,在本研究中,我们旨在评估罗格列酮治疗是否在炎症途径之外有作用,特别是对我们模型中的抗氧化途径。
对怀孕的C57BL/6J小鼠(胚胎期16.5天)用磷酸盐缓冲盐水(PBS)、罗格列酮(Rosi)、脂多糖(LPS;200μL 1XPBS中含10μg)或LPS + Rosi(LPS注射后6小时)进行处理。收集子宫肌层和蜕膜组织,并使用磁珠细胞分选和F4/80 +抗体进行巨噬细胞分离处理。通过定量聚合酶链反应对抗氧化因子Nrf2和Ho-1以及LPS受体Tlr4的表达水平进行定量。通过免疫荧光染色评估蛋白质水平。
与PBS对照组相比,LPS处理动物的蜕膜和子宫肌层巨噬细胞中Ho-1和Nrf2的表达显著降低,Tlr4的表达更高。与LPS组相比,LPS + Rosi组动物的巨噬细胞中Ho-1和Nrf2的表达显著升高,Tlr4的表达下调。
给予罗格列酮可通过下调炎症和上调抗氧化反应来预防PTB。