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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(duì)D-半乳糖致MSCs衰老的保護(hù)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(duì)D-半乳糖致間充質(zhì)干細(xì)胞（mesenchymal stem cells，MSCs）衰老的保護(hù)作用，并探究其潛在機(jī)制。方法 分組實(shí)驗(yàn)。正常對(duì)照組，采用完全培養(yǎng)基對(duì)MSCs進(jìn)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(duì)MSCs進(jìn)行48 h衰老誘導(dǎo)培養(yǎng)；HSYA組，在D-半乳糖致衰基礎(chǔ)上，采用合適濃度的HSYA對(duì)MSCs進(jìn)行保護(hù)。采用化學(xué)比色法、β-半乳糖苷酶染色法、EdU摻入法和流式細(xì)胞術(shù)對(duì)與氧化應(yīng)激、炎癥反應(yīng)、細(xì)胞衰老、細(xì)胞增殖與凋亡緊密相關(guān)的關(guān)鍵指標(biāo)進(jìn)行檢測(cè)；并采用Western blotting檢測(cè)與細(xì)胞增殖、細(xì)胞衰老和NF-κB通路活性密切關(guān)聯(lián)的蛋白的相對(duì)含量。結(jié)果 與衰老組相比，利用120 mg/L的HSYA對(duì)MSCs進(jìn)行保護(hù)能顯著改善其不良狀況。即顯著減輕了MSCs內(nèi)因D-gal誘導(dǎo)產(chǎn)生的氧化應(yīng)激和炎癥反應(yīng)；MSCs衰老率明顯下降；通過(guò)上調(diào)細(xì)胞增殖核抗原（PCNA）表達(dá)和抑制p16表達(dá)促進(jìn)了MSCs的增殖；通過(guò)下調(diào)cleaved Caspase-3和Bax的相對(duì)含量抑制了MSCs的衰老；并通過(guò)抑制IKKβ和p65發(fā)生磷酸化反應(yīng)下調(diào)了NF-κB通路的活性。結(jié)論 采用120 mg/L 的HSYA進(jìn)行干預(yù)能有效延緩D-gal所致的MSCs體外衰老進(jìn)程，其機(jī)制可能與HSYA的抗炎癥和抗氧化功效及其對(duì)NF-κB通路活性的抑制作用相關(guān)。

    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(guān)鍵詞:
    細(xì)胞衰老;羥基紅花黃色素;間充質(zhì)干細(xì)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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