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| 六味地黄丸通过SLC7A11/GPX4抑制MC3T3-E1细胞铁死亡的机制 |
| Mechanism of Liuwei Dihuang Pill in inhibiting ferroptosis of MC3T3-E1 cells through the SLC7A11/GPX4 |
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| DOI：10.3969/j.issn.1006-7108.2026.06.003 |
| 中文关键词: 骨质疏松症 铁死亡 六味地黄丸 成骨分化 SLC7A11/GPX4通路 |
| 英文关键词:Osteoporosis Ferroptosis Liuwei Dihuang Pill Osteogenic differentiation SLC7A11/GPX4 pathway |
| 基金项目:国家自然科学基金项目（82374483，82274563）；福建省自然科学基金项目（2022J01863） |
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| 中文摘要: |
| 目的 探讨六味地黄丸在抑制Erastin诱导的MC3T3-E1细胞铁死亡的作用及其机制。方法 用Erastin处理MC3T3-E1细胞构建铁死亡模型。20 %六味地黄丸含药血清干预 MC3T3-E1细胞，将细胞分为正常对照组（无Erastin）、模型组（Erastin）、空白血清组（Erastin+空白血清）、六味地黄丸组（Erastin+六味地黄丸含药血清）、抑制剂组（Erastin+六味地黄丸含药血清+SLC7A11抑制剂HG106）。通过CCK-8法检测细胞活力；荧光探针H2DCFDA检测细胞活性氧（reactive oxygen spcies，ROS）水平；碱性磷酸酶（alkaline phosphatase，
ALP）检测成骨分化能力；qPCR 检测铁死亡相关基因 SLC7A11、GPX4、CHAC1、PTGS2及成骨分化相关基因RUNX2、OCN、OSX的mRNA水平；Western blot检测铁死亡相关蛋白 SLC7A11、GPX4、CHAC1、PTGS2及成骨分化相关蛋白RUNX2、OCN、OSX的蛋白水平。结果 六味地黄丸显著降低了Erastin处理的MC3T3-E1中ROS的水平，抗铁死亡基因SLC7A11、GPX4 mRNA和蛋白表达水平显著提高，促铁死亡基因CHAC1、PTGS2 mRNA和蛋白的表达水平显著下降，六味地黄丸还上调了ALP活性以及成骨相关基因RUNX2、OCN和OSX的mRNA和蛋白水平。这些效应可通过信号通路抑制剂HG106逆转。结论 六味地黄丸通过SLC7A11/GPX4通路抑制MC3T3-E1细胞铁死亡并促进成骨分化。 |
| 英文摘要: |
| Objective To explore the effect and mechanism of Liuwei Dihuang Pill in inhibiting erastin-induced ferroptosis of MC3T3-E1 cells. Methods A ferroptosis model was established using erastin-treated MC3T3-E1 cells. MC3T3-E1 cells were treated with 20% Liuwei Dihuang-containing serum.The cells were divided into five groups: normal control (no erastin), model (erastin), blank serum (erastin + blank serum), Liuwei Dihuang Pill (erastin + Liuwei Dihuang Pill-containing serum), and inhibitor (erastin + 20% Liuwei Dihuang Pill-containing serum + 2.5 μM SLC7A11 inhibitor HG106). Cell viability was assessed by CCK-8 assay; intracellular reactive oxygen species (ROS) levels were measured using the H2DCFDA fluorescent probe; alkaline phosphatase (ALP) activity was evaluated to determine osteogenic differentiation capacity; mRNA levels of ferroptosis-related genes (SLC7A11, GPX4, CHAC1, PTGS2) and osteogenic differentiation genes (RUNX2, OCN, OSX) were quantified by qPCR; protein expression of these genes was analyzed by Western blot. Results Liuwei Dihuang Pill significantly reduced ROS levels in erastin-treated MC3T3-E1 cells, upregulated mRNA and protein expression of anti-ferroptosis genes (SLC7A11, GPX4), and downregulated pro-ferroptosis genes (CHAC1, PTGS2). Additionally, Liuwei Dihuang Pill enhanced ALP activity and increased mRNA and protein levels of osteogenic genes (RUNX2, OCN, OSX). These protective effects were reversed by the SLC7A11 inhibitor HG106. Conclusion Liuwei Dihuang Pill inhibits ferroptosis and promotes osteogenic differentiation in MC3T3-E1 cells via the SLC7A11/GPX4 pathway. |
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