[关键词]
[摘要]
目的 探讨竹节参总皂苷对妊娠期高血压疾病胎盘滋养细胞氧化应激反应与自噬的影响及作用机制。方法 采用100 μmol/L亚硝基左旋精氨酸甲酯(N-nitro-L-arginine methylester,L-NAME)处理人胎盘滋养细胞株HTR-8/SV-neo 48 h,并给予竹节参总皂苷(20、40、80 μg/mL)干预24 h,采用CCK-8法检测各组细胞活力;采用流式细胞仪检测各组细胞凋亡情况;采用试剂盒检测各组细胞上清液中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽(glutataione,GSH)水平;采用qRT-PCR和Western blotting法检测各组细胞脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)mRNA和蛋白表达情况;采用Western blotting法检测各组细胞微管相关蛋白1轻链3-II(microtubule associated protein 1 light chain 3 II,LC3-II)、LC3-I、p62和自噬效应蛋白(Beclin-1)表达情况;采用免疫荧光法检测各组细胞LC3表达情况。结果 与模型组比较,竹节参总皂苷组细胞活力显著升高(P<0.05),细胞凋亡率显著降低(P<0.01);上清液中SOD和GSH水平显著升高(P<0.01),MDA水平显著降低(P<0.01);FABP4 mRNA和蛋白表达水平显著降低(P<0.05);LC3-Ⅱ和Beclin-1蛋白表达水平显著降低(P<0.05),p62蛋白表达水平显著升高(P<0.05);LC3荧光染色强度降低。结论 竹节参总皂苷能够改善妊娠期高血压疾病胎盘滋养细胞中的氧化应激反应,并抑制过度自噬,其机制可能与调控FABP4表达有关。
[Key word]
[Abstract]
Objective To investigate the effect and mechanism of total saponins from Panax japonicus on oxidative stress response and autophagy in placental trophoblast cells of pregnancy-induced hypertension. Methods N-nitro-L-arginine methylester (L-NAME, 100 μmol/L) was used to treat human placental trophoblast cell line HTR-8/SV-neo for 48 h, and total saponins from P. japonicus (20, 40, 80 μg/mL) were used to intervene for 24 h, cell viability of each group was detected by CCK-8 method; Apoptosis of each group was detected by flow cytometry; Levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) in supernatant of each group was detected by kits; qRT-PCR and Western blotting were used to detect fatty acid binding protein 4 (FABP4) mRNA and protein expression; Western blotting was used to detect expressions of microtubule associated protein 1 light chain 3 II (LC3-II), LC3-I, p62 and autophagy effector protein (Beclin-1) in each group of cells; Immunofluorescence method was used to detect the LC3 expression in each group of cells. Results Compared with model group, cell viability in total saponins from P. japonicus group was significantly increased (P < 0.05), and apoptosis rate was significantly reduced (P < 0.01); Levels of SOD and GSH in supernatant were significantly increased (P < 0.01), MDA level was significantly reduced (P < 0.01); FABP4 mRNA and protein expression levels were significantly reduced (P < 0.05); LC3-Ⅱ and Beclin-1 protein expression levels were significantly reduced (P < 0.05), p62 protein expression levels were significantly increased (P < 0.05); Intensity of LC3 fluorescence staining was reduced. Conclusion Total saponins from P. japonicus can improve the oxidative stress response in placental trophoblasts of hypertensive disorders of pregnancy and inhibit excessive autophagy, mechanism of which may be related to the regulation of FABP4 expression.
[中图分类号]
R285.5
[基金项目]
河南省医学科技攻关计划项目(LHGJ20190609)