李 华,侯春霞,王俊芝,刘玉强,肖克源.隐丹参酮通过调节SDF-1/CXCR4信号通路保护人骨髓间充质干细胞免受过氧化氢诱导的细胞凋亡[J].,2024,(14):2639-2644 |
隐丹参酮通过调节SDF-1/CXCR4信号通路保护人骨髓间充质干细胞免受过氧化氢诱导的细胞凋亡 |
Cryptotanshinone Protects Human Bone Marrow Mesenchymal Stem Cells from Hydrogen Peroxide Induced Cell Apoptosis by Regulating the SDF-1/CXCR4 Signaling Pathway |
投稿时间:2023-12-27 修订日期:2024-01-23 |
DOI:10.13241/j.cnki.pmb.2024.14.007 |
中文关键词: 隐丹参酮 人骨髓间充质干细胞 SDF-1/CXCR4信号通路 氧化应激 细胞凋亡 |
英文关键词: Cryptotanshinone Human bone marrow mesenchymal stem cells SDF-1/CXCR4 signaling pathway Oxidative stress Cell apoptosis |
基金项目:山西省卫生健康委科研项目(2023141) |
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中文摘要: |
摘要 目的: 探讨隐丹参酮调节基质细胞衍生因子-1(SDF-1)/C-X-C型趋化因子受体4(CXCR4)信号通路抑制过氧化氢诱导的人骨髓间充质干细胞(BMSC)凋亡的作用机制。方法:体外培养人骨髓间充质干细胞(HBMSC),利用H2O2建立氧化应激模型。将HBMSC细胞分为对照组(不进行氧化应激诱导)、模型组(500 μmol/L H2O2诱导24 h)、隐丹参酮低剂量组(10 μmol/L)、隐丹参酮中剂量组(20 μmol/L)、隐丹参酮高剂量组(40 μmol/L)。细胞计数试剂盒(CCK-8)检测细胞活性;流式细胞术检测细胞凋亡;酶联免疫吸附法(Elisa)检测细胞葡萄糖摄取量和乳酸水平;微量法检测细胞中谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)含量;蛋白质印迹法(Western blot)检测细胞中半胱氨酸天冬氨酸蛋白酶3(caspase-3)、B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、SDF-1、CXCR4蛋白表达情况。结果:与对照组比较,模型组HBMSC细胞葡萄糖摄取量和乳酸水平、GSH、SOD活性、Bcl-2蛋白表达、SDF-1/CXCR4信号通路蛋白表达下降,细胞凋亡率、凋亡蛋白caspase-3、Bax表达、MDA含量上升(P<0.05);与模型组比较,隐丹参酮低、中、高剂量组HBMSC细胞葡萄糖摄取量和乳酸水平、GSH、SOD活性、Bcl-2蛋白表达、SDF-1/CXCR4信号通路蛋白表达上升,细胞凋亡率、凋亡蛋白caspase-3、Bax表达、MDA含量下降(P<0.05),且隐丹参酮低、中、高剂量组各指标差异具有统计学意义(P<0.05)。结论:隐丹参酮可能通过促进SDF-1/CXCR4信号通路转导,抑制过氧化氢诱导的BMSC氧化应激损伤,抑制其凋亡,其中高剂量隐丹参酮的效果最好。 |
英文摘要: |
ABSTRACT Objective: To investigate the mechanism by cryptotanshinone regulates the signaling pathway of stromal cell derived factor 1 (SDF-1)/C-X-C chemokine receptor 4 (CXCR4) and inhibits hydrogen peroxide induced cell apoptosis of human bone marrow mesenchymal stem cells (BMSCs). Methods: Cultivated human bone marrow mesenchymal stem cells (HBMSCs) in vitro and established an oxidative stress model by H2O2. Divided HBMSC cells into control group (without oxidative stress induction) and model group (500 μmol/L H2O2 induction for 24 hours), low-dose group of cryptotanshinone(10 μmol/L), medium dose group of cryptotanshinone (20 μmol/L), high-dose group of cryptotanshinone(40 μmol/L). Cell activity was detected by Cell Count Kit (CCK-8). Cell apoptosis was detected by flow cytometry detection. Cellular glucose uptake and lactate levels was detected by enzyme linked immunosorbent assay(Elisa) testing. The content of glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MDA) in cells were detected by micro method. Cysteine aspartate protease 3(caspase-3), B-lymphoblastoma-2 associated X protein (Bax), B-lymphoblastoma-2 (Bcl-2), expression of SDF-1 and CXCR4 proteins was detected by western blot detection (Western blot). Results: Compared with the control group, the glucose uptake and lactate levels, GSH, SOD activity, Bcl-2 protein expression, and SDF-1/CXCR4 signaling pathway protein expression of HBMSC cells in the model group decreased, while the cell apoptosis rate, expression of apoptotic protein caspase-3, Bax, and MDA content increased(P<0.05). Compared with the model group, the low, medium, and high dose groups of cryptotanshinone showed an increase in glucose uptake and lactate levels, GSH, SOD activity, Bcl-2 protein expression, and SDF-1/CXCR4 signaling pathway protein expression in HBMSC cells, while the cell apoptosis rate, expression of apoptotic proteins caspase-3, Bax, and MDA content decreased (P<0.05). The differences in various indicators among the low, medium, and high dose groups of cryptotanshinone were statistically significant(P<0.05). Conclusion: Cryptotanshinone may inhibit oxidative stress damage and apoptosis of BMSC induced by hydrogen peroxide by promoting SDF-1/CXCR4 signaling pathway transduction, with high-dose cryptotanshinone having the best effect. |
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