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Archive > Volume 15 Issue 1 > 2023,15(1):86-93. DOI:https://doi.org/10.1016/j.chmed.2022.06.011 Prev Next

羥基紅花黃色素對D-半乳糖致MSCs衰老的保護作用

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Protective effect of hydroxysafflor yellow A on MSCs against senescence induced by D-galactose

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    摘要:
    目的 研究羥基紅花黃色素（Hydroxysafflor yellow A，HSYA）對D-半乳糖致間充質干細胞（mesenchymal stem cells，MSCs）衰老的保護作用，并探究其潛在機制。方法 分組實驗。正常對照組，采用完全培養(yǎng)基對MSCs進行常規(guī)培養(yǎng)；衰老組，在完全培養(yǎng)基中添加D-半乳糖（D-gal，10 g/L），對MSCs進行48 h衰老誘導培養(yǎng)；HSYA組，在D-半乳糖致衰基礎上，采用合適濃度的HSYA對MSCs進行保護。采用化學比色法、β-半乳糖苷酶染色法、EdU摻入法和流式細胞術對與氧化應激、炎癥反應、細胞衰老、細胞增殖與凋亡緊密相關的關鍵指標進行檢測；并采用Western blotting檢測與細胞增殖、細胞衰老和NF-κB通路活性密切關聯(lián)的蛋白的相對含量。結果 與衰老組相比，利用120 mg/L的HSYA對MSCs進行保護能顯著改善其不良狀況。即顯著減輕了MSCs內因D-gal誘導產生的氧化應激和炎癥反應；MSCs衰老率明顯下降；通過上調細胞增殖核抗原（PCNA）表達和抑制p16表達促進了MSCs的增殖；通過下調cleaved Caspase-3和Bax的相對含量抑制了MSCs的衰老；并通過抑制IKKβ和p65發(fā)生磷酸化反應下調了NF-κB通路的活性。結論 采用120 mg/L 的HSYA進行干預能有效延緩D-gal所致的MSCs體外衰老進程，其機制可能與HSYA的抗炎癥和抗氧化功效及其對NF-κB通路活性的抑制作用相關。

    Abstract:
    Objective: To examine the protective effects of hydroxysafflor yellow A (HSYA) against the senescence of mesenchymal stem cells (MSCs) induced by D-galactose (D-gal) in vitro, and investigate the potential mechanism involved. Methods: Grouping experiment, Normal control (NC) group: conventional culture with complete medium; Senescence group: MSCs were cultured for 48 h with complete medium containing 10 g/L D-gal; HSYA group: on the basis of senescence induction, HSYA with the suitable concentration was used to protect MSCs. The key experimental indices associated with oxidative stress, inflammatory response, cell senescence, proliferation and apoptosis were measured through chemical colorimetry, b-galactosidase staining, EdU incorporation and flow cytometry, respectively. The relative quantity (RQ) of proteins related closely to cell proliferation, apoptosis, and NF-jB signaling were measured by Western blotting. Results: As compared with Senescence group, treatment with HSYA (120 mg/L) effectively ameliorated the adverse situation of MSCs. Oxidation stress and inflammation along with D-Gal induction was dramatically alleviated in MSCs; The b-Gal-positive staining indicated that MSC senescence was significantly mitigated; The proliferative capability of MSCs was significantly increased by up-regulating PCNA and inhibiting p16 expression; The anti-apoptotic effect on MSCs was exerted by down-regulating the RQ of cleaved Caspase-3 and Bax; The activity of NF-jB signaling in MSCs was notably suppressed through inhibiting phosphorylation of IKKb and p65. Conclusion: HSYA (120 mg/L) significantly delayed the D-Gal-induced senescence process in MSCs through attenuating inflammatory reaction and oxidative stress, and suppressing the activity of NF-jB signaling.

    關鍵詞:
    細胞衰老;羥基紅花黃色素;間充質干細胞

    Keywords:


    Project Supported:
    This study was supported by Applied Basic Research Project of Zhangjiakou Science and Technology Bureau (No. 1911020D), General Project of Hebei North University (No. YB2018034), Morphology Experimental Teaching Center of Hebei Province (No. XTZX201901) and College Students’ Innovation and Entrepreneurship Training Program (No. S202110092005).

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