目的 探究光甘草定衍生物MG01對β-淀粉樣多肽1-42（beta-amyloid peptide 1-42，Aβ_1-42）誘導的小鼠海馬神經(jīng)元細胞株HT22鐵死亡的保護以及預防作用。方法 計算機模擬分子對接驗證MG01可以與雌激素受體β（etrogen receptor beta，ERβ）以及過氧化物酶體增殖物激活受體γ共激活因子-1α‌‌（peroxisome proliferator-activated receptor-γ coactivator-1α，PGC-1α）相互作用；利用Cell counting kit-8（CCK-8）試劑盒分析MG01對細胞活力的影響并確定最佳給藥濃度；活性氧（reactive oxygen species，ROS）試劑盒檢測細胞ROS的水平；JC-1線粒體膜電位試劑盒檢測線粒體膜電位；細胞免疫熒光染色檢測神經(jīng)元樹突標志物微管相關蛋白-2（microtubule-associated protein-2，MAP-2）、鐵死亡特異性標志物過氧化物酶3（peroxiredoxin 3，PRDX3）的變化；蛋白質免疫印跡（Western blotting，WB）法檢測ERβ、ERα‌‌、谷胱甘肽過氧化物酶4（glutathion peroxidase，GPX4）、PRDX3、膜鐵轉運蛋白1（ferroportin1，F(xiàn)PN1）、鐵蛋白（ferritin）、PGC-1α、核因子E2相關因子2‌（nuclear factor erythroid 2-related factor 2，NRF2）、線粒體轉錄因子A（transcription factor A，TFAM）蛋白表達水平。結果 WB結果表明MG01能顯著上調(diào)ERβ與PGC-1α的表達（P＜0.05、0.01）；CCK-8、細胞形態(tài)檢測與免疫熒光結果表明，與模型組比較，MG01組細胞活力和形態(tài)得到明顯改善（P＜0.05、0.01），且有效濃度顯著低于光甘草定（P＜0.05）。WB結果表明，MG01能夠改善Aβ_1-42誘導的細胞鐵死亡，改善鐵死亡標志蛋白GPX4、PRDX3、FPN1以及ferritin的表達（P＜0.01），其作用機制可能與上調(diào)ERβ/PGC-1α促進線粒體生物發(fā)生，改善線粒體膜電位降低，降低ROS的水平有關。結論 MG01介導ERβ/PGC-1α促進線粒體生物發(fā)生，改善線粒體功能，抑制鐵死亡，發(fā)揮神經(jīng)保護作用。

Objective To investigate the protective and preventive effects of glabridin derivative MG01 on ferroptosis induced by beta-amyloid peptide 1-42 (Aβ1-42) in HT22 cells. Methods Computer simulation of molecular docking verifies that MG01 can interact with etrogen receptor beta (ERβ) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α). Cell counting kit-8 (CCK-8) was used to analyze the effects of MG01 on cell viability and determine the optimal concentration of MG01. Reactive oxygen species (ROS) detection kit was used to detect the ROS level of cells, and JC-1 mitochondrial membrane potential detection kit was used to detect mitochondrial membrane potential. Cell immunofluorescence (ICC) was used to detect the changes of neuronal dendrite marker microtubule-associated protein-2 (MAP-2), ferroptosis specific marker peroxiredoxin 3 (PRDX3). Western blotting (WB) was used to detect the expression levels of ERβ, ERα‌, glutathion peroxidase (GPX 4), PRDX3, ferroportin1 (FPN1), ferritin, PGC-1α, nuclear factor erythroid 2-related factor 2(NRF2), and transcription factor A (TFAM). Results WB results showed that MG01 could significantly increase the expression of ERβ and PGC-1α (P < 0.05, 0.01). CCK-8, cell morphology detection and immunofluorescence results showed that compared with the model group, the cell viability and morphology of the MG01 group were significantly improved (P < 0.05, 0.01), and the effective concentration was significantly lower than that of glabridin (P < 0.05). WB results showed that MG01 could improve the cell ferroptosis induced by Aβ_1-42, improve the expression of ferroptosis marker proteins GPX4, PRDX3, FPN1, and ferritin (P < 0.01), and its mechanism may be related to upregulating ERβ/PGC-1α to promote mitochondrial biogenesis, improving mitochondrial membrane potential, and reducing the level of ROS. Conclusion MG01 mediates ERβ/PGC-1α to promote mitochondrial biogenesis, improve mitochondrial function, inhibit ferroptosis, and play a neuroprotective role.

R285.5

山東省自然科學基金項目（ZR2024MH226）;國家自然科學基金項目（82073827）