天山茶藨对脂肪间充质干细胞成脂成骨分化的影响OACSTPCD
Effects of Ribes meyeri extracts on adipogenic and osteogenic differentiation of adipose-derived stem cells
目的 探究天山茶藨(Ribes meyeri)对脂肪间充质干细胞(adipose-derived stem cells,ADSCs)成脂成骨分化的影响.方法 对p9和p17代次的ADSCs进行成脂成骨的定向分化,通过β-半乳糖苷酶染色、荧光定量聚合酶链式反应(real-time PCR)测定细胞中P16INK4a、P53基因表达来检测细胞衰老情况;采用CCK-8法检测天山茶藨对ADSCs的细胞毒性,并利用细胞内活性氧类(reactive oxygen species,ROS)试剂盒检测天山茶藨对细胞内ROS含量的影响;将天山茶藨处理的衰老细胞进行成脂定向分化,年轻细胞进行成骨定向分化,并检测细胞中成脂标志基因LPL、PPARγ和成骨标志基因OCN、RUNX2、ALP的基因表达量;利用未分化、成脂成骨分化和天山茶藨处理的成脂成骨分化细胞构建加权基因共表达网络图谱(weighted gene co-expression network analysis,WGCNA);对天山茶藨处理的衰老细胞成脂分化和年轻细胞成骨分化中差异表达基因进行KEGG富集.结果 p17代次较之p9代次ADSCs细胞衰老程度加重,且衰老的ADSCs更易向成脂细胞分化,年轻的ADSCs则更易向成骨细胞分化;0.1 pg/μL天山茶藨促进ADSCs的细胞增殖,1 ng/mL和100pg/mL天山茶藨降低ADSCs中ROS水平;天山茶藨抑制衰老ADSCs的成脂分化,促进年轻ADSCs的成骨分化;衰老和年轻ADSCs具有不同的基因表达模式,天山茶藨在成脂分化过程中改变衰老ADSCs的表达模式;天山茶藨对ADSCs成脂成骨分化的影响与TGF-β、精氨酸生物合成信号通路相关.结论 天山茶藨促进年轻ADSCs的成骨分化,抑制衰老ADSCs的成脂细胞.
Objective To explore the effect of Ribes meyeri extracts on adipogenic and osteogenic differentiation of adipose-derived stem cells(ADSCs).Methods ADSCs of the p9 and p17 generations were used in the study,cell senescence was detected by β-galactosidase staining and expression of P16INK4a and P53 determined by real-time PCR.The cytotoxicity of Ribes meyeri on ADSCs was detected by CCK-8 assay,and the effect of Ribes meyeri on intracellular ROS content was detected by ROS kit.ADSCs of the p17 generation(senescent cells)treated with Ribes meyeri were subjected to adipogenic differentiation and ADSCs of the p9 generation(young cells)treated with Ribes meyeri were subjected to osteogenic differentiation.The expression of the adipogenic marker genes LPL and PPARγ,and the osteogenic marker genes OCN,RUNX2,and ALP were detected.Weighted gene co-expression network analysis(WGCNA)was constructed using undifferentiated,adipogenic and osteogenic differentiated and Ribes meyeri-treated adipogenic and osteogenic differentiated cells,respectively.KEGG enrichment of differentially expressed genes was performed for adipogenic differentiation of senescent cells and osteogenic differentiation of young cells treated with Ribes meyeri.Results The p17 generation of ADSCs was more senescent than p9 generations.The senescent ADSCs were more likely to differentiate into adipogenic cells,while young ADSCs were more likely to differentiate into osteoblasts.Ribes meyeri extract at concentration of 0.1 pg/μL promoted the cell proliferation of ADSCs,and Ribes meyeri at concentration of 1 ng/mL and 100 pg/mL decreased the level of ROS in ADSCs.Ribes meyeri inhibited the adipogenic differentiation of senescent ADSCs,and promoted osteogenic differentiation.The gene expression patterns of senescent and young ADSCs were different,and the gene expression pattern of young ADSCs was altered during the process of adipogenic differentiation.The effects of Ribes meyeri on adipogenic and osteogenic differentiation of ADSCs were correlated with TGF-β and Arg biosynthesis signaling pathways.Conclusion Ribes meyeri promotes the osteogenic differentiation of young ADSCs and inhibits the adipogenic differentiation of senescent ADSCs.
刘青;苏婷婷;高家明;刘海亮
同济大学医学院,上海 200331石河子大学生命科学学院新疆植物药资源利用教育部重点实验室,新疆石河子 832003同济大学医学院,上海 200331||石河子大学生命科学学院新疆植物药资源利用教育部重点实验室,新疆石河子 832003||同济大学附属东方医院干细胞转化医学产业基地,上海 200123
药学
天山茶藨脂肪间充质干细胞成脂成骨分化
Ribes meyeriadipose mesenchymal stem cellsadipogenic and osteogenic differentiation
《同济大学学报(医学版)》 2024 (004)
471-479 / 9
国家自然科学基金(82271593)
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