Tantengco Ourlad Alzeus G, Vink Joy, Medina Paul Mark B, Menon Ramkumar
Division of Maternal-Fetal Medicine and Perinatal Research, Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA.
Biological Models Laboratory, Department of Biochemistry and Molecular Biology, College of Medicine, University of the Philippines Manila, Manila, Philippines.
Biol Reprod. 2021 Jul 2;105(1):204-216. doi: 10.1093/biolre/ioab058.
A physiologic increase in reactive oxygen species throughout pregnancy is required to remodel the cervix. Oxidative stress can cause cellular damage that contributes to dysfunctional tissue. This study determined the oxidative stress-induced cell fate of human cervical epithelial and cervical stromal cells. We treated the ectocervical and endocervical epithelial cells and cervical stromal cells with cigarette smoke extract, an oxidative stress inducer, for 48 h. Cell viability (crystal violet assay); cell cycle, apoptosis, and necrosis (flow cytometry); senescence (senescence-associated β-galactosidase staining); autophagy (staining for autophagosome protein, microtubule-associated protein 1 light chain 3B); stress signaler p38 mitogen-activated protein kinases pathway activation (western blot analyses); and inflammation by measuring interleukin-6 (enzyme-linked immunosorbent assay) were conducted after 48 h of cigarette smoke extract treatment. Oxidative stress induced reactive oxygen species production in cervical cells, which was inhibited by N-acetylcysteine. Oxidative stress promoted cell cycle arrest and induced necrosis in cervical cells. High senescence and low autophagy were observed in cervical stromal cells under oxidative stress. Conversely, senescence was low and autophagy was high in endocervical epithelial cells. Oxidative stress induced p38 mitogen-activated protein kinases (p38MAPK) activation in all cervical cells but only increased interleukin-6 production by the ectocervical epithelial cells. Inhibition of interleukin-6 production by a p38 mitogen-activated protein kinases inhibitor confirmed the activation of an oxidative stress-induced pathway. In conclusion, oxidative stress can promote cell death and sterile inflammation that is mediated by p38 mitogen-activated protein kinases activation in the cellular components of the cervix. These cellular damages may contribute to the normal and premature cervical ripening, which can promote preterm birth.
整个孕期活性氧的生理性增加对于宫颈重塑是必需的。氧化应激可导致细胞损伤，进而导致组织功能障碍。本研究确定了氧化应激诱导的人宫颈上皮细胞和宫颈基质细胞的细胞命运。我们用香烟烟雾提取物（一种氧化应激诱导剂）处理宫颈外口和宫颈内口上皮细胞以及宫颈基质细胞48小时。在香烟烟雾提取物处理48小时后，进行细胞活力（结晶紫测定）、细胞周期、凋亡和坏死（流式细胞术）、衰老（衰老相关β-半乳糖苷酶染色）、自噬（自噬体蛋白微管相关蛋白1轻链3B染色）、应激信号蛋白p38丝裂原活化蛋白激酶途径激活（蛋白质印迹分析）以及通过测量白细胞介素-6进行炎症检测（酶联免疫吸附测定）。氧化应激诱导宫颈细胞产生活性氧，而N-乙酰半胱氨酸可抑制这种产生。氧化应激促进宫颈细胞的细胞周期停滞并诱导坏死。在氧化应激下，宫颈基质细胞中观察到高衰老和低自噬。相反，宫颈内口上皮细胞中衰老程度低而自噬程度高。氧化应激在所有宫颈细胞中诱导p38丝裂原活化蛋白激酶（p38MAPK）激活，但仅增加宫颈外口上皮细胞的白细胞介素-6产生。p38丝裂原活化蛋白激酶抑制剂抑制白细胞介素-6产生证实了氧化应激诱导途径的激活。总之，氧化应激可促进细胞死亡和无菌性炎症，这是由宫颈细胞成分中p38丝裂原活化蛋白激酶激活介导的。这些细胞损伤可能导致正常和过早的宫颈成熟，进而促进早产。
