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王光祖,王少敏,乔素兰,李晓敬,姚春霞,布兰娜.芹菜素调节AMPK/mTOR通路对子痫前期胎盘滋养细胞自噬的影响研究[J].湖南中医药大学学报,2023,43(8):1394-1401[点击复制] |
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| 芹菜素调节AMPK/mTOR通路对子痫前期胎盘滋养细胞自噬的影响研究 |
| 王光祖,王少敏,乔素兰,李晓敬,姚春霞,布兰娜 |
| (邯郸市妇幼保健院, 邯郸市儿童医院 新生儿科, 河北 邯郸 056000;河北工程大学医学院, 河北 邯郸 056038) |
| 摘要: |
| 目的 探究芹菜素对子痫前期（preeclampsia，PE）胎盘滋养细胞自噬的影响及可能的作用机制。方法 取人绒毛膜滋养层HTR-8/Svneo细胞，采用缺氧复氧1 h（hypoxic-reoxygenation 1 h，H1R1）诱导建立PE细胞模型，并通过MTT法检测芹菜素对HTR-8/Svneo细胞毒性的影响。取对数生长期的HTR-8/Svneo细胞，分为对照组、H1R1组、腺苷酸活化蛋白激酶（adenosine monophosphate-activated protein kinase，AMPK）激活剂阿卡地新（acadesine，AICAR）组（1 mmol/L）、芹菜素低剂量组（10 μmol/L）、芹菜素高剂量组（20 μmol/L）、芹菜素+AICAR组（芹菜素20 μmol/L+AICAR 1 mmol/L）。采用MTT法检测细胞活力；透射电子显微镜观察细胞自噬情况；划痕实验和Transwell小室实验检测细胞迁移、侵袭能力；Western blot法检测细胞AMPK/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）通路和自噬、侵袭相关蛋白的表达。结果 与对照组比较，10、20 μmol/L芹菜素对HTR-8/Svneo细胞增殖有促进作用（P<0.05）。与对照组比较，H1R1组细胞在24、48、72 h时细胞活力、划痕愈合率和穿膜细胞数、N-钙黏蛋白（N-cadherin）表达、p-mTOR/mTOR比值显著降低（P<0.05），微管相关蛋白1轻链3-Ⅱ（microtubule-associated protein 1 light chain 3-Ⅱ，LC3Ⅱ）/微管相关蛋白1轻链3-Ⅰ（microtubule-associated protein 1 light chain 3-Ⅰ，LC3Ⅰ）比值、Beclin-1和E-钙黏蛋白（E-cadherin）表达、p-AMPK/AMPK比值显著升高（P<0.05），自噬小体数量增多（P<0.05）；与H1R1组比较，芹菜素低、高剂量组细胞活力、划痕愈合率和穿膜细胞数、N-cadherin表达、p-mTOR/mTOR比值显著升高（P<0.05），LC3II/LC3I比值、Beclin-1和E-cadherin表达、p-AMPK/AMPK比值显著降低（P<0.05），自噬小体数量减少（P<0.05）；与芹菜素高剂量组比较，芹菜素+AICAR组细胞在24、48、72 h时细胞活力、划痕愈合率和穿膜细胞数、N-cadherin表达、p-mTOR/mTOR比值显著降低（P<0.05），LC3II/LC3I比值、Beclin-1和E-cadherin表达、p-AMPK/AMPK比值显著升高（P<0.05），自噬小体数量增多（P<0.05）。结论 芹菜素能增强滋养细胞的侵袭能力，改善PE，其作用机制可能与抑制AMPK/mTOR通路介导的胎盘滋养细胞过度自噬有关。 |
| 关键词: 芹菜素 子痫前期 胎盘滋养细胞 自噬 迁移 侵袭 腺苷酸活化蛋白激酶 哺乳动物雷帕霉素靶蛋白 |
| DOI：10.3969/j.issn.1674-070X.2023.08.008 |
| 投稿时间：2023-05-22 |
| 基金项目:河北省自然科学基金项目（H2020402004）；邯郸市科学技术研究与发展计划项目（19422083040ZC）。 |
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| Effects of apigenin on autophagy of placental trophoblast cells in preeclampsia by regulating AMPK/mTOR pathway |
| WANG Guangzu,WANG Shaomin,QIAO Sulan,LI Xiaojing,YAO Chunxia,BU Lanna |
| (Department of Neonatology, Handan Maternal and Child Health Hospital & Handan Children's Hospital, Handan, Hebei 056000, China;School of Medicine, Hebei University of Engineering, Handan, Hebei 056038, China) |
| Abstract: |
| Objective To explore the effects of apigenin on autophagy of placental trophoblast cells in preeclampsia (PE) and its possible mechanism of action. Methods Human chorionic trophoblast HTR-8/Svneo cells were taken and hypoxia-reoxygenation 1 h (H1R1) was used to induce the establishment of PE cell models. The effects of apigenin on HTR-8/Svneo cytotoxicity was tested by MTT method. In addition, HTR-8/Svneo cells in logarithmic growth phase were taken and divided into control group, H1R1 group, adenosine monophosphate-activated protein kinase (AMPK) activator acadesine (AICAR) group (1 mmol/L), low-dose apigenin group (10 μmol/L), high-dose apigenin group (20 μmol/L), and apigenin+AICAR group (apigenin 20 μmol/L+AICAR 1 mmol/L). Cell viability was determined by MTT method; cell autophagy was observed by transmission electron microscope; cell migration and invasion ability were tested by scratch assay and Transwell chamber assay. Western blot method was used to examine the related protein expressions of cellular AMPK/mammalian target of rapamycin (mTOR) pathway, autophagy, and invasion. Results Compared with the control group, 10 and 20 μmol/L apigenin had a promoting effect on the proliferation of HTR-8/Svneo cells (P<0.05). Compared with the control group, the cell viability at 24, 48, and 72 h, scratch healing rate, number of transmembrane cells, N-cadherin expression, and p-mTOR/mTOR ratio of the H1R1 group were significantly lower (P<0.05), while the microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ)/microtubule-associated protein 1 light chain 3 Ⅰ(LC3Ⅰ) ratio, Beclin-1 and E-cadherin expressions, p-AMPK/AMPK ratio were significantly higher (P<0.05), and the number of autophagosomes increased (P<0.05); compared with the H1R1 group, the cell viability at 24, 48, and 72 h, scratch healing rate, number of transmembrane cells, N-cadherin expression, and p-mTOR/mTOR ratio in low- and high-dose apigenin groups were significantly elevated (P<0.05), while LC3II/LC3I ratio, Beclin-1 and E-cadherin expressions, and p-AMPK/AMPK ratio were significantly reduced (P<0.05), and the number of autophagosomes became lower (P<0.05); compared with high-dose apigenin group, the cell viability at 24, 48 and 72 h, scratch healing rate, number of transmembrane cells, N-cadherin expression, and p-mTOR/mTOR ratio of apigenin+AICAR group were significantly lower (P<0.05), while LC3II/LC3I ratio, Beclin-1 and E-cadherin expressions, and p-AMPK/AMPK ratio were significantly higher (P<0.05), and the number of autophagosomes increased (P<0.05). Conclusion Apigenin can enhance the invasion ability of trophoblast cells and thus improve PE, and its mechanism of action may be related to the inhibition of the excessive autophagy of placental trophoblast cells mediated by AMPK/mTOR pathway. |
| Key words: apigenin preeclampsia placental trophoblast cells autophagy migration invasion adenosine monophosphate-activated protein kinase mammalian target of rapamycin |
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