摘要
目的 探讨高糖高脂环境诱导骨髓间充质干细胞(BMSCs)损伤及其与铁死亡的相关性.方法 将BMSCs分为正常组(5.50 mmol·L-1葡萄糖)和高糖高脂组(25.00 mmol·L-1葡萄糖+0.25 mmol·L-1棕榈酸).用β-半乳糖苷酶染色评估细胞衰老情况;用酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子-o(TNF-α)、白细胞介素-10(IL-10)水平;用试剂盒检测谷胱甘肽(GSH)、丙二醛(MDA)、亚铁离子(Fe2+);用蛋白质印迹法检测酰基辅酶A合成酶长链家族成员4(ACSL4)、花生四烯酸15脂氧合酶(ALOX15)、谷胱甘肽过氧化物酶4(GPX4)的蛋白表达水平.结果 正常组、高糖高脂组的细胞衰老比率分别为(6.80±1.60)％和(13.00±1.58)％,TNF-α 含量分别为(122.54±3.94)和(169.77±2.89)pg·mL-1,IL-10 含量分别为(155.16±3.97)和(105.15±7.30)pg·mL-1,GSH 含量分别为 4.30±0.33 和 1.55±0.14,MDA含量分别为2.94±0.10和5.84±0.10,Fe2+含量分别为6.22±0.35和16.13±0.36,ACSL4蛋白相对表达水平分别为0.42±0.05和0.84±0.10,ALOX15蛋白相对表达水平分别为0.61±0.25和1.06±0.11,GPX4蛋白相对表达水平分别为1.13±0.17和0.33±0.08.高糖高脂组的上述指标与正常组比较,在统计学上差异均有统计学意义(均P＜0.01).结论 高糖高脂诱导BM-SCs的损伤,在这一过程中铁死亡扮演了重要角色.
Abstract
Objective To investigate the relationship between the injury and ferroptosis of bone marrow mesenchymal stem cells(BMSCs)induced by high glucose and high fat.Methods BMSCs were divided into normal group(5.50 mmol·L-1 glucose)and high glucose and high fat(HGHF)group(25.00 mmol·L-1 glucose+0.25 mmol·L-1 palmitic acid).Assessment of cellular aging via β-galactosidase staining;enzyme linked immunosorbent assay(ELISA)were used to detect tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)release levels;glutathione(GSH),malondialdehyde(MDA)and ferrous ion(Fe2+)detection kits were used to detect ferroptosis related indicators;Western blotting was used to detect the expression of ferroptosis related signaling pathway protein acyl-CoA synthetase long chain family member 4(ACS14)/arachidonate 15-lipoxygenase(ALOX15)/glutathione peroxidase 4(GPX4).Results The senescence rates of normal group HGHF group were(6.80±1.60)％and(13.00±1.58)％;the levels of TNF-α were(122.54±3.94)and(169.77±2.89)pg·mL-1;the levels of IL-10 were(155.16±3.97)and(105.15±7.30)pg·mL-1;GSH levels were 4.30±0.33 and 1.55±0.14;MDA levels were 2.94±0.10 and 5.84±0.10;Fe2+levels were 6.22±0.35 and 16.13±0.36;the relative expression levels of ACSL4 protein were 0.42±0.05 and 0.84±0.10;the relative ALOX15 protein were 0.61±0.25 and 1.06±0.11;the relative expression levels of GPX4 protein were 1.13±0.17 and 0.33±0.08,respectively.The above indexes in the HGHF group were significantly different from those in the normal group(all P＜0.05).Conclusion 25 mmol·L-1 glucose combined with 0.25 mmol·L-1 palmitic acid for 24 h can be used as a suitable condition to induce BMSCs injury.ferroptosis plays an important role in BMSCs injury induced by high glucose and high fat.
基金项目
国家自然科学基金资助项目(81760767)
新疆天然药物活性组分与释药技术重点实验室基金资助项目(XJDX1713)
国家科技期刊平台
NSTL
万方数据