This study explored the therapeutic effect of bone marrow mesenchymal stem cell‐derived exosomes on the treatment of obesity‐induced fracture healing. Quantitative real‐time PCR was used to detect the expression of lncRNA H19, miR‐467 and Hoxa10 and combined with WB detection to detect osteogenic markers (RUNX2, OPN, OCN). Determine whether exosomes have entered BMSCs by immunofluorescence staining. Alkaline phosphatase (ALP) and alizarin red staining (ARS) staining were used to detect ALP activity and calcium deposition. We found that high‐fat treatment can inhibit the secretion of BMSCs‐derived exosomes and affect the expression of H19 carried by them. In vivo and in vitro experiments show that high‐fat or obesity factors can inhibit the expression of osteogenic markers and reduce the staining activity of ALP and ARS. The treatment of exosomes from normal sources can reverse the phenomenon of osteogenic differentiation and abnormal fracture healing. Further bioinformatics analysis found that miR‐467 as a regulatory molecule of lncRNA H19 and Hoxa10, and we verified the targeting relationship of the three through dual luciferase report experiments. Further, we found similar phenomena in ALP and ARS staining. Bone marrow mesenchymal stem cell‐derived exosomes improve fracture healing caused by obesity.

中文翻译：

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肥胖通过 BMSC 衍生的外泌体 LncRNA H19 调节 miR-467/HoxA10 轴对成骨分化和骨折愈合

本研究探讨了骨髓间充质干细胞衍生的外泌体对肥胖诱导骨折愈合的治疗作用。采用定量实时PCR检测lncRNA H19、miR-467和Hoxa10的表达，并结合WB检测检测成骨标志物（RUNX2、OPN、OCN）。通过免疫荧光染色确定外泌体是否已进入骨髓间充质干细胞。碱性磷酸酶（ALP）和茜素红染色（ARS）染色用于检测ALP活性和钙沉积。我们发现高脂肪处理可以抑制 BMSCs 来源的外泌体的分泌并影响其携带的 H19 的表达。体内外实验表明，高脂肪或肥胖因子可以抑制成骨标志物的表达，降低ALP和ARS的染色活性。对正常来源的外泌体进行治疗可以逆转成骨分化和骨折异常愈合的现象。进一步的生物信息学分析发现miR-467作为lncRNA H19和Hoxa10的调控分子，我们通过双荧光素酶报告实验验证了三者的靶向关系。此外，我们在 ALP 和 ARS 染色中发现了类似的现象。骨髓间充质干细胞衍生的外泌体改善肥胖引起的骨折愈合。