The combination of mitogenic stimulation and DNA damage induces chondrocyte senescence.
The combination of mitogenic stimulation and DNA damage induces chondrocyte senescence.
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DOI：
10.1016/j.joca.2020.11.004
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发表时间：
2021-03
影响因子：
7
通讯作者：
Diekman BO
中科院分区：
文献类型：
--
作者：
Copp ME;Flanders MC;Gagliardi R;Gilbertie JM;Sessions GA;Chubinskaya S;Loeser RF;Schnabel LV;Diekman BO
细胞衰老是一种表型状态，其特征在于稳定的细胞周期停滞、增强的溶酶体活性以及炎性分子和基质降解酶的分泌。衰老与骨关节炎（OA）病理生理学有关;然而，驱动软骨和其他关节组织衰老诱导的机制尚不清楚。虽然许多生理信号能够启动衰老，但一个新出现的主题是受损细胞在持续的促有丝分裂刺激下转化为衰老。本研究的目的是建立一种体外关节软骨移植模型，以探讨衰老诱导的机制。本研究使用了来自尸体马后膝关节和人踝关节的健康软骨。照射外植体以引发DNA损伤，并通过含血清培养基和转化生长因子β1和碱性成纤维细胞生长因子处理提供促有丝分裂刺激。衰老的读数是定量流式细胞术测定以检测衰老相关的β半乳糖苷酶活性（SA-β-gal）、p16和γ H2 AX的免疫荧光以及炎症基因表达的qPCR。与基线对照条件相比，人软骨外植体需要辐照和促有丝分裂刺激来诱导衰老（7.16% vs 2.34% SA-β-gal高，p=0.0007）。这些条件还导致外植体内的软骨细胞簇、持续的DNA损伤反应、p16增加和基因表达变化。在细胞损伤的背景下，用促有丝分裂刺激物处理软骨外植体可靠地诱导高水平的SA-β-gal活性和其他衰老标记物，这提供了生理学相关的模型系统来研究衰老诱导的机制。
Cellular senescence is a phenotypic state characterized by stable cell-cycle arrest, enhanced lysosomal activity, and the secretion of inflammatory molecules and matrix degrading enzymes. Senescence has been implicated in osteoarthritis (OA) pathophysiology; however, the mechanisms that drive senescence induction in cartilage and other joint tissues are unknown. While numerous physiological signals are capable of initiating senescence, one emerging theme is that damaged cells convert to senescence in response to sustained mitogenic stimulation. The goal of this study was to develop an in vitro articular cartilage explant model to investigate the mechanisms of senescence induction. This study utilized healthy cartilage derived from cadaveric equine stifles and human ankles. Explants were irradiated to initiate DNA damage, and mitogenic stimulation was provided through serum-containing medium and treatment with transforming growth factor β1 and basic fibroblastic growth factor. Readouts of senescence were a quantitative flow cytometry assay to detect senescence-associated β galactosidase activity (SA-β-gal), immunofluorescence for p16 and γH2AX, and qPCR for the expression of inflammatory genes. Human cartilage explants required both irradiation and mitogenic stimulation to induce senescence as compared to baseline control conditions (7.16% vs. 2.34% SA-β-gal high, p=0.0007). These conditions also resulted in chondrocyte clusters within explants, a persistent DNA damage response, increased p16, and gene expression changes. Treatment of cartilage explants with mitogenic stimuli in the context of cellular damage reliably induces high levels of SA-β-gal activity and other senescence markers, which provides a physiologically relevant model system to investigate the mechanisms of senescence induction.
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影响因子：
64.5
作者：
López-Otín C;Blasco MA;Partridge L;Serrano M;Kroemer G
通讯作者：
Kroemer G
DOI：
10.1146/annurev-pathol-121808-102144
发表时间：
2010
期刊：
Annual review of pathology
影响因子：
--
作者：
Coppé JP;Desprez PY;Krtolica A;Campisi J
通讯作者：
Campisi J
影响因子：
3.7
作者：
HAYFLICK, L;MOORHEAD, PS
通讯作者：
MOORHEAD, PS
影响因子：
5.3
作者：
Kurtz, Steven M.;Ong, Kevin L.;Bozic, Kevin J.
通讯作者：
Bozic, Kevin J.
影响因子：
4.1
作者：
Feng, Meng;Peng, Hang;Qiu, Yusheng
通讯作者：
Qiu, Yusheng