| 徐双双,邵玲玲,冯向东.五味子醇甲抑制肝细胞铁死亡保护对乙酰氨基酚肝损伤的研究[J].安徽医药,2025,29(11):2158-2164. |
| 五味子醇甲抑制肝细胞铁死亡保护对乙酰氨基酚肝损伤的研究 |
| Schizandrol A protects against acetaminophen-induced liver injury by inhibiting hepatocyte ferroptosis |
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| DOI：10.3969/j.issn.1009-6469.2025.11.008 |
| 中文关键词: 五味子 对乙酰氨基酚 铁死亡 药物性肝损伤 活性氧 |
| 英文关键词: Chinese magnoliavine fruit Acetaminophen Ferroptosis Drug-induced liver injury Reactive oxygen species |
| 基金项目:上海市药学会上海医院药学科研基金资助项目( 2023-YY-11) |
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| 中文摘要: |
| 目的研究五味子醇甲( schizandrol A，Sch A)对对乙酰氨基酚( APAP)肝损伤的保护作用及其机制。方法该研究起止时间为 2023年 10月至 2024年 4月。用细胞计数试剂盒( CCK-8)法检测 Sch A和 APAP对人正常肝细胞 LO2存活率的影响；将 LO2细胞分为对照组、模型组( APAP 10 mmol/L)、 Sch A低剂量组( APAP 10 mmol/L+Sch A 5 μmol/L)、 Sch A中剂量组( APAP 10 mmol/L+Sch A 10 μmol/L)、 Sch A高剂量组( APAP 10 mmol/L+Sch A 20 μmol/L)，用试剂盒检测细胞中还原型谷胱甘肽(GSH)水平，及细胞培养上清液中天冬氨酸氨基转移酶( AST)和丙氨酸氨基转移酶( ALT)含量；用 DCFH-DA荧光探针检测细胞内活性氧水平； C11-BODIPY荧光探针检测细胞内脂质过氧化水平；实时荧光 PCR(real-time PCR)和蛋白质印迹法分别检测溶质载体超家族 7成员 11(SLC7A11)谷胱甘肽过氧化物酶 4(GPX4)溶质载体超家族 40成员 1(SLC40A1)、转铁蛋白、核转录因子红系 2相关因子 2(Nrf2)、血红素加、氧酶 -1(HO-1)的 mRNA水平和蛋、白表达情况。结果 Sch A在 1.25~20.00 μmol/L时对 LO2细胞增殖无抑制作用，且 Sch A在 5、10、20 μmol/L时显示对 LO2细胞有一定的增殖促进作用。与模型组比较， Sch A(5、 10、20 μmol/L)能够显著提高细胞存活率［(66.47±4.81)%、(75.70±6.21)%、(85.24±5.40)%比( 50.44±4.70)%］、细胞内 GSH含量［( 28.10±1.97)%、(31.45±2.89)%、(40.45±4.89)%比( 19.20±2.38)%］，明显降低细胞培养上清液中 AST［( 21.10±2.11)%、(15.90±1.12)%、(11.00±0.92)%比( 26.35±1.58)%］和 ALT含量［( 75.70±6.21)%、(52.85±4.98)%、(39.15±5.21)%比( 89.95±4.82)%］(P<0.05)而且 Sch A(5、10、20 μmol/L)显著降低细胞内活性氧水平( 109.66±6.03、75.49±4.16、36.21±4.48比 125.29±7.74)(P<0.05)。型组比较， Sch A(5、10、20 μmol/L)显著缓解细胞铁死亡，明显降低细胞内脂质过氧化水平( 2.38±0.11、与模，1.85±0.21、1.33±0.17比 3.00±0.15)、细胞内铁含量( 4.25±0.20、3.09±0.26、2.11±0.14比 5.88±0.33)(细胞内脂质过氧化水平和细胞内铁含量为与对照组相对倍数值)，明显减少细胞乳酸脱氢酶( LDH)的泄漏［( 360.67±23.71)U/g、(280.67±17.79)U/g、(219.67±17.56)U/g比( 476.67±21.03)U/g)］(P<0.05)；而且 Sch A(5、10、20 μmol/L)使铁死亡相关基因 SLC7A11(0.41±0.10、 0.62±0.04、0.89±0.09比 0.27±0.07)、 GPX4(0.49±0.08、0.59±0.06、0.89±0.08比 0.29±0.06)的 mRNA水平显著升高，使 SLC7A11(0.47±0.06、0.74±0.08、0.89±0.04比 0.34±0.06)、 GPX4(0.51±0.04、0.64±0.06、0.85±0.06比 0.37±0.05)的蛋白表达显著增多( P<0.05)。与模型组比较， Sch A(5、10、20 μmol/L)显著升高 Nrf2(0.51±0.04、0.73±0.08、0.88±0.10比 0.29±0.08)和 HO-1(0.54±0.08、0.68±0.06、0.85±0.08比 0.34±0.06)的 mRNA水平，使 Nrf2(0.49±0.04、0.65±0.09、0.77±0.11比 0.36±0.06)和 HO-1(0.47±0.08、0.64±0.08、0.82±0.04比 0.35±0.05)的蛋白表达显著增多( P<0.05)。结论 Sch A明显改善 APAP导致的肝细胞损伤，其机制可能与抑制肝细胞铁死亡有关。 |
| 英文摘要: |
| Objective To investigate the protective effect of schizandrol A (Sch A) against acetaminophen (APAP)-induced liver inju-ry and its underlying mechanism. Methods This study was conducted from October 2023 to April 2024. The Cell Counting Kit-8 (CCK-8) assay was used to evaluate the effects of Sch A and APAP on the viability. LO2 cells were divided into the following groups:control group, model group (APAP 10 mmol/L), and Sch A low-, medium-, and high-dose groups (APAP 10 mmol/L+Sch A 5, 10, 20μmol/L). The levels of reduced glutathione (GSH) in the cells and the contents of aspartate aminotransferase (AST) and glutamate ami-notransferase (ALT) in the supernatant of cell culture were measured using commercial kits. Intracellular reactive oxygen species (ROS)levels were detected using the DCFH-DA fluorescent probe, and lipid peroxidation levels were measured using the C11-BODIPY fluo-rescent probe. Real-time PCR and Western blotting were used to determine the mRNA levels and protein expressions of solute carrierfamily 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), solute carrier family 40 member 1 (SLC40A1), transferrin, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1), respectively.Results Sch A at concentrations ranging from1.25 to 20.00 μmol/L did not inhibit LO2 cell proliferation and even promoted proliferation at 5, 10, and 20 μmol/L. Compared with themodel group, Sch A (5, 10, 20 μmol/L) significantly increased cell viability [(66.47±4.81)%, (75.70±6.21)%, (85.24±5.40)% vs. (50.44± 4.70)%], intracellular GSH content [(28.10±1.97)%, (31.45±2.89)%, (40.45±4.89)% vs. (19.20±2.38)%], while markedly reduced AST [(21.10±2.11)%, (15.90±1.12)%, (11.00±0.92)% vs. (26.35±1.58)%] and ALT levels [(75.70±6.21)%, (52.85±4.98)%, (39.15±5.21)% vs. (89.95±4.82)%] in the cell culture supernatant ( P<0.05). Furthermore, Sch A (5, 10, 20 μmol/L) significantly decreased intracellu-lar ROS levels (109.66±6.03, 75.49±4.16, 36.21±4.48 vs. 125.29±7.74) (P<0.05). Compared with the model group, Sch A (5, 10, 20μmol/L) significantly alleviated ferroptosis, reduced intracellular lipid peroxidation (2.38±0.11, 1.85±0.21, 1.33±0.17 vs. 3.00±0.15), decreased intracellular iron content (4.25±0.20, 3.09±0.26, 2.11±0.14 vs. 5.88±0.33) (the levels of intracellular lipid peroxidation andintracellular iron content are relative multiples of the control group), and reduced lactate dehydrogenase (LDH) leakage [(360.67±23.71) U/g, (280.67±17.79) U/g, (219.67±17.56) U/g vs. (476.67±21.03) U/g] (P<0.05). Moreover, Sch A (5, 10, 20 μmol/L) significantly increased the mRNA levels of ferroptosis-related genes SLC7A11 (0.41±0.10, 0.62±0.04, 0.89±0.09 vs. 0.27±0.07) and GPX4 (0.49± 0.08, 0.59±0.06, 0.89±0.08 vs. 0.29±0.06), and enhanced the protein expression of SLC7A11 (0.47±0.06, 0.74±0.08, 0.89±0.04 vs. 0.34±0.06) and GPX4 (0.51±0.04, 0.64±0.06, 0.85±0.06 vs. 0.37±0.05) (P<0.05). Compared with the model group, Sch A (5, 10, 20μmol/L) significantly elevated the mRNA levels of Nrf2 (0.51±0.04, 0.73±0.08, 0.88±0.10 vs. 0.29±0.08) and HO-1 (0.54±0.08, 0.68± 0.06, 0.85±0.08 vs. 0.34±0.06), and increased the protein expression of Nrf2 (0.49±0.04, 0.65±0.09, 0.77±0.11 vs. 0.36±0.06) and HO-1 (0.47±0.08, 0.64±0.08, 0.82±0.04 vs. 0.35±0.05) (P<0.05), in a dose-dependent manner.Conclusion Sch A significantly ameliorates APAP-induced hepatocyte injury, and its mechanism may be associated with the inhibition of hepatocyte ferroptosis. |
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