Hattori Y, Kotani T, Tsuda H, Mano Y, Tu L, Li H, Hirako S, Ushida T, Imai K, Nakano T, Sato Y, Miki R, Sumigama S, Iwase A, Toyokuni S, Kikkawa F
Departments of Gynecology and Obstetrics, Nagoya University Graduate School of Medicine , Nagoya , Japan.
Free Radic Res. 2015;49(8):1026-37. doi: 10.3109/10715762.2015.1038257. Epub 2015 May 7.
Maternal inflammation is associated with spontaneous preterm birth and respiratory impairment among premature infants. Recently, molecular hydrogen (H2) has been reported to have a suppressive effect on oxidative stress and inflammation. The aim of this study was to evaluate the effects of H2 on fetal lung injury caused by maternal inflammation. Cell viability and the production of interleukin-6 (IL-6) and reactive oxygen species (ROS) were examined by treatment with lipopolysaccharide (LPS) contained in ordinal or H2-rich medium (HM) using a human lung epithelial cell line, A549. Pregnant Sprague Dawley rats were divided into three groups: Control, LPS, and HW + LPS groups. Rats were injected with phosphate-buffered saline (Control) or LPS intraperitoneally (LPS) on gestational day 19 and provided H2 water (HW) ad libitum for 24 h before LPS injection (HW + LPS). Fetal lung samples were collected on day 20, and the levels of apoptosis, oxidative damage, IL-6, and vascular endothelial growth factor (VEGF) were evaluated using immunohistochemistry. The number of apoptotic cells, and levels of ROS and IL-6 were significantly increased by LPS treatment, and repressed following cultured with HM in A549 cells. In the rat models, the population positive for cleaved caspase-3, 8-hydroxy-2'-deoxyguanosine, IL-6, and VEGF was significantly increased in the LPS group compared with that observed in the Control group and significantly decreased in the HW + LPS group. In this study, LPS administration induced apoptosis and oxidative damage in fetal lung cells that was ameliorated by maternal H2 intake. Antenatal H2 administration may decrease the pulmonary mobility associated with inflammation in premature infants.
母体炎症与早产及早产儿呼吸功能损害有关。最近，有报道称分子氢（H2）对氧化应激和炎症具有抑制作用。本研究的目的是评估H2对母体炎症所致胎儿肺损伤的影响。使用人肺上皮细胞系A549，通过在普通培养基或富氢培养基（HM）中加入脂多糖（LPS）进行处理，检测细胞活力以及白细胞介素-6（IL-6）和活性氧（ROS）的产生。将怀孕的Sprague Dawley大鼠分为三组：对照组、LPS组和HW + LPS组。在妊娠第19天，给大鼠腹腔注射磷酸盐缓冲盐水（对照组）或LPS，并在注射LPS前24小时随意给予H2水（HW + LPS组）。在第20天收集胎儿肺样本，使用免疫组织化学方法评估细胞凋亡、氧化损伤、IL-6和血管内皮生长因子（VEGF）的水平。LPS处理显著增加了凋亡细胞数量以及ROS和IL-6水平，而在A549细胞中用HM培养后这些指标受到抑制。在大鼠模型中，与对照组相比，LPS组中裂解的半胱天冬酶-3、8-羟基-2'-脱氧鸟苷、IL-6和VEGF的阳性细胞数量显著增加，而在HW + LPS组中显著减少。在本研究中，LPS给药诱导了胎儿肺细胞的凋亡和氧化损伤，而母体摄入H2可改善这种损伤。产前给予H2可能会降低早产儿与炎症相关的肺部易感性。
