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首页> 外文期刊>Journal of tissue engineering and regenerative medicine >Regulating effect of Circ_ATRNL1 on the promotion of SOX9 expression to promote chondrogenic differentiation of hAMSCs mediated by MiR-145-5p
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Regulating effect of Circ_ATRNL1 on the promotion of SOX9 expression to promote chondrogenic differentiation of hAMSCs mediated by MiR-145-5p

机译：Circ_ATRNL1对SOX9促进促进MIR-145-5P介导的HAMSCs软骨分化的调节效果

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摘要

Circ_ATRNL1 is significantly highly expressed in cartilage tissues of patients with osteoarthritis. This study explored the role and mechanism of circ_ATRNL1 in cartilage differentiation of human adipose-derived mesenchymal stem cells (hAMSCs). hAMSCs were isolated and identified by flow cytometry. The degree of chondrocyte and adipogenic differentiation was assessed using Alcian blue staining and Oil Red O staining, respectively. The expressions of cartilage- and adipogenic-related genes, circ_ATRNL1, and SOX9 were detected by reverse transcription quantitative polymerase chain reaction. The correlation between SOX9 and circ_ATRNL1 was analyzed using Pearson test. Bioinformatics and luciferase analysis were used to detect the overlapped target miRNAs of circ_ATRNL1 and SOX9. The role of circ_ATRNL1/miRNA/SOX9 was examined using functional rescue assays. hAMSCs were identified as CD90-, CD105-, and CD44-positive. The degree of cartilage differentiation of hAMSCs was significantly enhanced after 2 weeks. Cartilage-related genes, circ_ATRNL1 and SOX9, were significantly upregulated, and positively correlated with each other. Circ_ATRNL1 overexpression enhanced hAMSC proliferation and differentiation into chondrogenesis, and promoted the expressions of COL2, Aggrecan, and SOX9. Overexpression of circ_ATRNL1 inhibited the adipogenic differentiation of hAMSCs and the expressions of adipogenic-related genes. miR-145-5p was a target miRNA for circ_ATRNL1 and SOX9. miR-145-5p mimic inhibited hAMSC differentiation toward cartilage, and inhibited the expression of cartilage-related factors. miR-145-5p mimic effectively reversed the regulating effect of circ_ATRNL1 on hAMSCs. Circ_ATRNL1 regulates the promotion of SOX9 expression to promote chondrogenic differentiation of hAMSCs mediated by miR-145-5p.
机译：Circ_ATRNL1在骨关节炎患者的软骨组织中显著高表达。本研究探讨了circ_ATRNL1在人脂肪间充质干细胞（HAMSC）软骨分化中的作用及其机制。用流式细胞术分离鉴定hAMSCs。软骨细胞和脂肪细胞的分化程度分别采用阿尔辛蓝染色和油红O染色进行评估。通过逆转录定量聚合酶链反应检测软骨和脂肪生成相关基因、circ_ATRNL1和SOX9的表达。采用皮尔逊检验分析SOX9与circ_ATRNL1之间的相关性。生物信息学和荧光素酶分析用于检测circ_ATRNL1和SOX9的重叠靶miRNA。使用功能性拯救分析检测circ_ATRNL1/miRNA/SOX9的作用。hAMSCs被鉴定为CD90、CD105和CD44阳性。2周后，hAMSCs的软骨分化程度显著增强。软骨相关基因circ_ATRNL1和SOX9显著上调，并且彼此呈正相关。Circ_ATRNL1的过度表达增强了hAMSC的增殖和向软骨形成的分化，并促进了COL2、聚集蛋白聚糖和SOX9的表达。circ_ATRNL1的过度表达抑制了火腿干细胞的成脂分化和成脂相关基因的表达。miR-145-5p是circ_ATRNL1和SOX9的靶向miRNA。miR-145-5p模拟物抑制hAMSC向软骨的分化，并抑制软骨相关因子的表达。miR-145-5p模拟物可有效逆转circ_ATRNL1对HAMSC的调节作用。Circ_ATRNL1调节SOX9表达的促进，以促进由miR-145-5p介导的HAMSC的软骨分化。

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