Department of Anesthesiology, Fujian Maternity and Child Health Hospital, 18 Daoshan Road, Gulou District, Fuzhou, 350001, Fujian, China.
Arch Gynecol Obstet. 2022 Apr;305(4):1055-1067. doi: 10.1007/s00404-021-06316-z. Epub 2022 Jan 4.
The purpose of this study was to evaluate the effect of propofol on lipopolysaccharide (LPS)-induced toxicity in HTR-8/SVneo cells.
In this study, HTR-8/SVneo cells were induced by LPS. The cells were treated with different concentrations of propofol. Cell proliferation, apoptosis, invasion, and wound healing rate were measured by MTT, flow cytometry, Transwell, and wound-healing assay. The relative mRNA expression levels of miR-216a-5p, TLR, MyD88, and NF-κB(p65) were measured by qRT-PCR. The protein expression levels of TLR, MyD88, and p-NF-κB(p65) were detected by western blot. The p-NF-κB(p65) nuclear volume was evaluated by cell immunofluorescence.
Compared with control group, the cell proliferation, invasion, and wound healing rate were significantly decreased and the cell apoptosis rate was significantly increased in LPS- induced HTR-8/SVneo cells (P < 0.01). With propofol supplement, the cell proliferation, migration, and invasion abilities were significantly recovered and apoptosis rate was significantly inhibited (P < 0.05). The expression levels of miR-216a-5p, TLR4, MyD88, NF-κB(p65), and p-NF-κB(p65), and p-NF-κB(p65) nuclear volume were significantly different between propofol group and model group (P < 0.05). However, after knockdown of miR-216a-5p expression by si-miR-216a-5p transfection, the cell proliferation, migration, and invasion abilities were significantly inhibited and apoptosis rate was notably increased (P < 0.05).
Propofol improves LPS-induced toxicity in HTR-8/SVneo cells via regulation miR-216a-5p/TLR4 axis.
本研究旨在评估异丙酚对脂多糖（LPS）诱导的 HTR-8/SVneo 细胞毒性的影响。
本研究采用 LPS 诱导 HTR-8/SVneo 细胞，用不同浓度的异丙酚处理细胞。通过 MTT、流式细胞术、Transwell 和划痕愈合试验测定细胞增殖、凋亡、侵袭和伤口愈合率。采用 qRT-PCR 测定 miR-216a-5p、TLR、MyD88 和 NF-κB(p65)的相对 mRNA 表达水平。采用 Western blot 检测 TLR、MyD88 和 p-NF-κB(p65)的蛋白表达水平。采用细胞免疫荧光法评估 p-NF-κB(p65)核体积。
与对照组相比，LPS 诱导的 HTR-8/SVneo 细胞的细胞增殖、侵袭和伤口愈合率明显降低，细胞凋亡率明显升高（P<0.01）。用异丙酚补充后，细胞增殖、迁移和侵袭能力明显恢复，凋亡率明显抑制（P<0.05）。异丙酚组与模型组之间 miR-216a-5p、TLR4、MyD88、NF-κB(p65)和 p-NF-κB(p65)的表达水平和 p-NF-κB(p65)核体积均有显著差异（P<0.05）。然而，转染 si-miR-216a-5p 抑制 miR-216a-5p 表达后，细胞增殖、迁移和侵袭能力明显受到抑制，凋亡率明显增加（P<0.05）。
异丙酚通过调节 miR-216a-5p/TLR4 轴改善 LPS 诱导的 HTR-8/SVneo 细胞毒性。
