Objective To observe the effect of SBI-0206965,a selective inhibitor of unc-51-like kinase (ULK) 1,on autophagy and related functions of HTR-8/Svneo cells under hypoxia/reoxygenation. Methods A hypoxia/reoxygenation culture model of trophoblast HTR-8/Svneo cells was established to simulate the pathophysiological state of oxidative stress in preeclampsia in vitro.The HTR-8/Svneo cells were divided into the normal control group,the hypoxia/reoxygenation group and the observation group (the hypoxic/reoxygenated cells treated with 10 μM SBI-0206965).The ROS activity was detected by the glossy sperm method,the MDA levels were detected by the thiobarbituric acid method,the cell viability was detected by CCK-8 kit,the expression of Beclin1 and LC3B was detected by real-time quantitative PCR,cell migration was detected by scratching test,and cell invasion ability was detected by Transwell chamber assay. Results Compared with the control group,the ROS activity (t=8.418,P<0.001),MDA content (t=5.788,P<0.001),Beclin1 mRNA expression (t=8.987,P<0.001) and LC3B mRNA expression (t=10.000,P<0.001) were significantly increased,while cell viability(t=4.747,P<0.001),cell invasion (t=5.923,P<0.001) and cell migration (t=12.306,P<0.001) were significantly decreased in the hypoxia/reoxygenation group.Compared with the hypoxia/reoxygenation group,the ROS activity (t=2.641,P=0.019),MDA content (t=2.216,P=0.043),Beclin1 mRNA expression (t=3.245,P=0.005) and LC3B mRNA expression (t=3.194,P=0.006) were significantly decreased,while cell viability (t=2.234,P=0.041),cell invasion (t=2.847,P=0.012) and cell migration (t=4.549,P<0.001) were significantly increased in the observation group. Conclusion ULK1 inhibitor SBI-0206965 can down-regulate the autophagy level of HTR-8/Svneo cells under oxidative stress in preeclampsia,and improve the activity and function of the damaged cells.
Key words
Preeclampsia /
Trophoblasts /
SBI-0206965 /
Autophagy /
Oxidative stress
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Liu Y,Li DY,Bolatai A,et al.Progress in research on biomarkers of gestational diabetes mellitus and preeclampsia[J].Diabetes Metab Syndr Obes,2023,16:3807-3815.
[2] Klionsky DJ,Petroni G,Amaravadi RK,et al.Autophagy in major human diseases[J].EMBO J,2021,40(19):e108863.
[3] Nakashima A,Tsuda S,Kusabiraki T,et al.Current understanding of autophagy in pregnancy[J].Int J Mol Sci,2019,20(9):2342.
[4] 冯晓玲,姜天悦,张杨.胎盘自噬与子痫前期[J].中国计划生育学杂志,2022,30(1):236-240.
[5] 彭婷婷,刘海燕,肖璐,等.重度子痫前期患者胎盘组织中Rab23的表达及对人胎盘滋养细胞凋亡及自噬的影响[J].中国医药,2021,16(3):431-435.
[6] Ren H,Dai R,Nik Nabil WN,et al.Unveiling the dual role of autophagy in vascular remodelling and its related diseases[J].Biomed Pharmacother,2023,168:115643.
[7] Li J,Zhang T,Ren T,et al.Oxygen-sensitive methylation of ULK1 is required for hypoxia-induced autophagy[J].Nat Commun,2022,13(1):1172.
[8] Diceglie C,Anelli GM,Martelli C,et al.Placental antioxidant defenses and autophagy-related genes in maternal obesity and gestational diabetes mellitus[J].Nutrients,2021,13(4):1303.
[9] Egan DF,Chun MG,Vamos M,et al.Small molecule inhibition of the autophagy kinase ULK1 and identification of ULK1 substrates[J].Mol Cell,2015,59(2):285-297.
[10] Lu J,Zhu L,Zheng LP,et al.Overexpression of ULK1 represents a potential diagnostic marker for clear cell renal carcinoma and the antitumor effects of SBI-0206965[J].EBioMedicine,2018,34:85-93.
[11] Hwang DY,Eom JI,Jang JE,et al.ULK1 inhibition as a targeted therapeutic strategy for FLT3-ITD-mutated acute myeloid leukemia[J].J Exp Clin Cancer Res,2020,39(1):85.
[12] Tang F,Hu P,Yang Z,et al.SBI0206965,a novel inhibitor of Ulk1,suppresses non-small cell lung cancer cell growth by modulating both autophagy and apoptosis pathways[J].Oncol Rep,2017,37(6):3449-3458.
[13] 王光祖,王少敏,乔素兰,等.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.
[14] 孙艺娟,贾杰,邓恋,等.丙泊酚通过miR-182-5p介导HIF-1α通路对缺氧诱导胎盘滋养细胞生物学活性的影响[J].实用医学杂志,2023,39(15):1869-1875.
[15] Chen G,Li J,Wang Z,et al.Ezetimibe protects against spinal cord injury by regulating autophagy and apoptosis through inactivation of PI3K/AKT/mTOR signaling[J].Am J Transl Res,2020,12(6):2685-2694.
[16] 刘丽霞,张路,杨丽.胃癌组织中miR-184、Beclin1、LC3B的表达及临床意义[J].山东医药,2021,61(13):29-32.
[17] Gu S,Zhou C,Pei J,et al.Esomeprazole inhibits hypoxia/endothelial dysfunction-induced autophagy in preeclampsia[J].Cell Tissue Res,2022,388(1):181-194.
[18] Yang D,Ding J,Wang Y,et al.YY1-PVT1 affects trophoblast invasion and adhesion by regulating mTOR pathway-mediated autophagy[J].J Cell Physiol,2020,235(10):6637-6646.
[19] 董会月,路君,林灵婧,等.ULK1复合物在细胞自噬中的作用[J].中华细胞与干细胞杂志(电子版),2016,6(4):240-242.
[20] 张岚,欧阳亮,刘博.自噬启动因子ULK1的生物学功能与靶向治疗研究进展[J].厦门大学学报(自然科学版),2019,58(6):787-801.
[21] Deng R,Zhang HL,Huang JH,et al.MAPK1/3 kinase-dependent ULK1 degradation attenuates mitophagy and promotes breast cancer bone metastasis[J].Autophagy,2021,17(10):3011-3029.
PDF(1016 KB)
