KAT7促进软骨细胞衰老
KAT7 promotes the aging of cartilage cellular senescence
王旭蕾 1储柱平 1王惠敏 1魏伟 1严尚学1
作者信息
- 1. 安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,抗炎免疫药物协同创新中心,合肥 230032
- 折叠
摘要
目的 建立过度复制诱导原代小鼠软骨细胞衰老模型及小鼠自然衰老模型,研究KAT7在软骨细胞及组织衰老中的作用.方法 二型胶原酶消化法获取小鼠膝关节软骨细胞,使用甲苯胺蓝染色及Col Ⅱ染色鉴定小鼠软骨细胞;Western blot和细胞免疫荧光法检测不同代次小鼠软骨细胞衰老信号相关蛋白及KAT7表达水平,SA-β-Gal染色确定细胞衰老情况.取22月龄老龄小鼠与2月龄年轻小鼠关节HE染色、番红O-固绿染色比较小鼠关节病理情况;免疫组化观察小鼠关节组织中KAT7和p53的表达情况.结果 与对照组相比,衰老的小鼠软骨细胞KAT7蛋白及p21、p53表达水平显著升高,KAT7抑制剂WM-3835可抑制衰老软骨细胞KAT7和p21的蛋白表达.SA-β-Gal染色可见,第八代(P8)较第一代(P1)软骨细胞阳染明显增加.与年轻小鼠软骨组织相比,老龄小鼠软骨组织呈现近骨性分布,软骨表面完整性降低,肥大软骨细胞数量显著增多,KAT7和p53细胞阳染增加.结论 KAT7在软骨细胞衰老模型及老龄小鼠软骨组织中表达升高,提示KAT7可能与软骨细胞衰老相关.
Abstract
Objective To investigate the role of KAT7 in cartilage cell and tissue ageing by establishing an over-replicating-induced primary mouse cartilage cell ageing model and a mouse natural ageing model.Methods Chon-drocytes of the mouse knee joint were obtained by type Ⅱ collagenase digestion and identified by toluidine blue stai-ning and Col Ⅱ staining.The age-related proteins and KAT7 expression levels in cartilage cells from different gener-ations of mice were discovered using Western blot and cellular immunofluorescence techniques,and the aging of the cells was assessed by SA-β-Gal coloring.The pathological alterations were examined in the joints of 22-month-old mice compared to 2-month-old mice using HE staining and safranin O-solid green staining.Additionally,immuno-histochemical analysis was done to observe the expression of KAT7 and p53 in mouse joint tissue.Results Com-pared with the control group,the expression levels of KAT7 protein and p21 and p53 in aged mouse chondrocytes significantly increased.WM-3835,a commonly inhibitor of KAT7 that possess the capacity on halting the protein expression procedure of gene KAT7 as well as p21 in ageing chondrocytes.SA-β-Gal staining showed a significant increase in positive staining of chondrocytes in the eighth generation(P8)compared to the first generation(P1).Compared with the cartilage tissue of young mice,the cartilage tissue of elderly mice presents a near-bone distribu-tion,with a decrease in cartilage surface integrity,a significant increase in the number of hypertrophic chondro-cytes,and more KAT7 and p53 cells that were positive.Conclusion The expression of KAT7 increases in the ageing chondrocytes and the cartilage tissue of ageing mice,reveales the potential significance of KAT7 correlated to cellular aging process in cartilage.
关键词
KAT7/衰老/软骨细胞Key words
KAT7/senescence/chondrocyte引用本文复制引用
基金项目
安徽省高等学校自然科学研究重大项目(KJ2020ZD15)
出版年
2024
国家科技期刊平台
NSTL
万方数据