摘要
目的探讨中性粒细胞胞外诱捕网(neutrophil extracellular traps, NETs)对人羊膜上皮细胞增殖和凋亡的影响。方法取正常妊娠足月择期剖宫产孕妇术前外周血, 体外提取中性粒细胞并诱导生成NETs。体外培养人羊膜上皮细胞(WISH细胞)并分为4组:(1)对照组:不经过任何处理;(2)NETs组:用NETs(500 ng/ml)刺激WISH细胞;(3)NETs+SB203580(p38激酶抑制剂)组:用SB203580(5 μmol/L)预处理WISH细胞30 min后加入NETs(500 ng/ml);(4)SB203580组:只加入SB203580。处理48 h后, 分别通过细胞增殖实验、乳酸脱氢酶(lactate dehydrogenase, LDH)检测实验和流式细胞技术检测各组细胞增殖比率、细胞上清LDH水平和细胞凋亡率。4组间比较采用单因素方差分析, 两两比较采用LSD-t检验。结果 (1)细胞增殖情况:NETs组细胞增殖比率低于对照组[(9.379±0.775)%与(36.560±1.208)%, LSD-t=20.78, P<0.001];NETs+SB203580组WISH细胞增殖比率[(27.920±0.926)%]高于NETs组(LSD-t=14.18, P<0.001)。(2)LDH:NETs组细胞上清LDH水平高于对照组(1.518±0.038与0.274±0.004, LSD-t=44.25, P<0.05), NETs+SB203580组LDH水平(0.857±0.009)低于NETs组(LSD-t=23.51, P<0.001)。(3)细胞凋亡情况:NETs组细胞凋亡率较对照组增加[(14.290±0.141)%与(10.110±0.044)%, LSD-t=21.76, P<0.001];NETs+SB203580组[(10.500±0.218)%]细胞凋亡率低于NETs组(LSD-t=19.70, P<0.001)。结论 NETs可能通过p38/丝裂原活化蛋白激酶信号通路抑制人羊膜上皮细胞的增殖, 并促进其凋亡。
Objective To investigate the effects of neutrophil extracellular traps(NETs)on the proliferation and apoptosis of human amniotic epithelial cells.Methods NETs were induced in vitro from the neutrophil cells obtained from the peripheral blood of normal pregnant women before elective cesarean section at full-term.Human amniotic epithelial cell lines(WISH cells)were cultured in vitro,and were divided into four groups:(1)control group:without any stimulus;(2)NETs group:WISH cells were stimulated with NETs(500 ng/ml);(3)NETs+SB203580(p38 kinase inhibitor)group:WISH cells were pretreated with SB203580(5μmol/L)for 30 min and then NETs(500 ng/ml)was added;(4)SB203580 group:only SB203580 was added.After stimulating for 48 h,cell proliferation assay,lactate dehydrogenase(LDH)assay,and flow cytometry assay were used to detect the cell proliferation rate,LDH level of cell supernatant,and cell apoptosis rate among different groups.The results were analyzed and compared using one-way analysis of variance and LSD-t test.Results(1)Cell proliferation:The cell proliferation ratio in the NETs group was lower than that in the control group[(9.379±0.775)%vs(36.560±1.208)%,LSD-t=20.78,P<0.001];and the figure in the NETs+SB203580 group[(27.920±0.926)%]was higher than that in the NETs group(LSD-t=14.18,P<0.001).(2)LDH:There was an increased LDH level in the cell supernatant of the NETs group compared with the control group(1.518±0.038 vs 0.274±0.004,LSD-t=44.25,P<0.05),and the LDH level in the NETs+SB203580 group(0.857±0.009)was decreased than that in the NETs group(LSD-t=23.51,P<0.001).(3)Apoptosis:Compared with the control group,the cell apoptosis level of the NETs group was increased[(14.290±0.141)%vs(10.110±0.044)%,LSD-t=21.76,P<0.001];but that in the NETs+SB203580 group[(10.500±0.218)%]was lower than in the NETs group(LSD-t=19.70,P<0.001).Conclusion p38/mitogen-activated protein kinases signaling pathway may be involved in the process of NETs,inhibiting proliferation and promoting apoptosis of human amniotic epithelial cells.
作者
胡敏
申迪
张美华
王谢桐
Hu Min;Shen Di;Zhang Meihua;Wang Xietong(Cheeloo College of Medicine,Shandong University,Jinan 250102,China;Department of Obstetrics,Shandong Provincial Hospital,Jinan 250021,China;Key Laboratory of Birth Regulation and Control Technology of National Health Commission of China,Jinan 250014,China;Department of Obstetrics,Shandong Maternal and Child Health Care Hospital,Jinan 250014,China)
出处
《中华围产医学杂志》
CAS
CSCD
北大核心
2022年第8期609-614,共6页
Chinese Journal of Perinatal Medicine
基金
国家自然科学基金(81873842)
山东省医药卫生科技发展计划(202105020991)。
