Qujing No.1 Hospital, Affiliated Qujing Hospital of Kunming Medical University, No. 1 Yuanlin Road, Qujing City, Yunnan Province, China.
Kunming Institute of Zoology, Chinese Academy of Sciences, No.17 Longxin Road, Kunming City, Yunnan Province, China.
Biomed Pharmacother. 2024 Oct;179:117349. doi: 10.1016/j.biopha.2024.117349. Epub 2024 Aug 26.
Adipose-derived mesenchymal stem cells (ADSCs) have received significant attention in the field of cartilage tissue repair. Angelica sinensis polysaccharide (ASP) can enhance both the proliferation and differentiation of mesenchymal stem cells. Therefore, we intend to explore the effect of ASP on chondrogenic differentiation of ADSCs in vitro, and elucidate the underlying mechanisms.
ADSCs were treated with different concentrations of ASP to determine the optimal concentration. The chondrogenic differentiation of ADSCs was evaluated using Alcian blue staining, qRT-PCR, western blot, and IF staining. Transcriptome sequencing was performed to identify the expression profiles of ADSCs before and after ASP treatment, followed by bioinformatic analyses including differential expression analysis, enrichment analysis, and construction of PPI networks to identify differentially expressed genes (DEGs) associated with ASP and chondrogenic differentiation.
Surface markers of isolated rat-derived ADSCs were identified by CD44CD90CD45CD106, and exhibited the capacity for lipogenic, osteogenic, and chondrogenic differentiation. With increasing concentration of ASP treatment, there was an upregulation in the activity and acidic mucosubstance of ADSCs. The levels of Aggrecan, COL2A1, and Sox9 showed an increase in ADSCs after 28 days of 80 µg/ml ASP treatment. Transcriptome sequencing revealed that ASP-associated DEGs regulate extracellular matrix synthesis, immune response, inflammatory response, and cell cycle, and are involved in the NF-κB, AGE-RAGE, and calcium pathways. Moreover, Edn1, Frzb, Mdk, Nog, and Sulf1 are hub genes in DEGs. Notably, ASP upregulated MDK levels in ADSCs, while knockdown of MDK mitigated ASP-induced elevations in acidic mucosubstance, chondrogenic differentiation-related markers (Aggrecan, COL2A1, and Sox9), and the activity of the PI3K/AKT pathway.
ASP enhances the proliferation and chondrogenic differentiation of ADSCs by activating the MDK-mediated PI3K/AKT pathway.
脂肪间充质干细胞（ADSCs）在软骨组织修复领域受到广泛关注。当归多糖（ASP）可促进间充质干细胞的增殖和分化。因此，我们旨在探讨 ASP 对体外 ADSC 软骨分化的影响，并阐明其潜在机制。
用不同浓度的 ASP 处理 ADSCs，以确定最佳浓度。通过阿尔辛蓝染色、qRT-PCR、western blot 和 IF 染色评估 ADSC 的软骨分化。进行转录组测序以鉴定 ASP 处理前后 ADSC 的表达谱，然后进行生物信息学分析，包括差异表达分析、富集分析和 PPI 网络构建，以鉴定与 ASP 和软骨分化相关的差异表达基因（DEGs）。
通过 CD44CD90CD45CD106 鉴定分离的大鼠源性 ADSC 的表面标志物，并表现出脂肪生成、成骨和成软骨分化的能力。随着 ASP 处理浓度的增加，ADSC 的活性和酸性黏多糖增加。在 80µg/ml ASP 处理 28 天后，ADSC 中 Aggrecan、COL2A1 和 Sox9 的水平增加。转录组测序显示，ASP 相关的 DEGs 调节细胞外基质合成、免疫反应、炎症反应和细胞周期，并参与 NF-κB、AGE-RAGE 和钙途径。此外，Edn1、Frzb、Mdk、Nog 和 Sulf1 是 DEGs 的枢纽基因。值得注意的是，ASP 上调了 ADSC 中的 MDK 水平，而 MDK 敲低则减轻了 ASP 诱导的酸性黏多糖、软骨分化相关标志物（Aggrecan、COL2A1 和 Sox9）以及 PI3K/AKT 通路活性的升高。
ASP 通过激活 MDK 介导的 PI3K/AKT 通路增强 ADSC 的增殖和软骨分化。
