Department of Human Anatomy, Research Center for Bone and Stem Cells; Key Laboratory for Aging & Disease, The State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.
Department of Nutrition and Food Safety, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China.
Stem Cells Dev. 2022 Sep;31(17-18):541-554. doi: 10.1089/scd.2021.0337. Epub 2022 Aug 22.
Alcohol consumption is regarded as one of the leading risk factors for secondary osteopenia. Angiogenesis and osteogenesis coupled by type-H vessels coordinate the biological process of bone homeostasis to prevent osteopenia. This study aimed to determine whether chronic alcohol inhibits type-H vessel-dependent bone formation. Two-month-old mice were fed with 5% (v/v) alcohol liquid diet (28% of calories) or normal liquid diet every day for 2 months. The tibias were isolated and detected with X-ray and microcomputed tomography. Paraffin-embedded or frozen tibial sections were prepared and used for immunohistochemical or immunofluorescence staining, respectively. Human umbilical vein endothelial cells (HUVECs) were treated with different concentrations of alcohol, including 0 mM (0%), 8.7 mM (0.5%), 52 mM (3%), or 87 mM (5%) alcohol for 12 h. The conditioned medium of the above HUVEC cells was collected to culture human bone marrow-mesenchymal stem cells (BM-MSCs), which were induced to differentiate into osteoblasts in vitro. The alcoholic diet retarded the bone growth and led to osteoporosis, impaired bone formation of osteoblasts, and decreased CD31EMCN type-H vessel formation through inhibiting proliferation and promoting aging of endothelial cells in mice. Alcohol treatment obviously increased the expression of p16, while significantly decreased the expression of Bmi-1, CDK6, Cyclin D, E2F1, and bone morphogenetic protein (BMP)2 compared with vehicle. Alcohol inhibited the differentiation of BM-MSCs into osteoblasts through reducing the BMP2 secretion of endothelial cells in type-H vessels. Alcoholic diet impaired CD31EMCN type-H vessel formation through inhibiting proliferation and promoting aging of endothelial cells through Bmi-1/p16 signaling, and inhibited the differentiation of BM-MSCs into osteoblasts through reducing the BMP2 secretion of endothelial cells in type-H vessels. This study provides a basis for developing a new treatment strategy targeting aging endothelial cells of type-H vessel to prevent alcoholic osteopenia.
饮酒被认为是导致继发性骨质疏松症的主要危险因素之一。由 H 型血管耦合的血管生成和成骨作用协调了骨稳态的生物学过程，以防止骨质疏松症。本研究旨在确定慢性酒精是否抑制 H 型血管依赖性骨形成。将 2 月龄的小鼠用 5%（体积/体积）酒精液体饮食（28%卡路里）或正常液体饮食每天喂养 2 个月。分离胫骨并用 X 射线和微计算机断层扫描进行检测。制备石蜡包埋或冷冻胫骨切片，分别用于免疫组织化学或免疫荧光染色。用人脐静脉内皮细胞（HUVEC）用不同浓度的酒精处理，包括 0 mM（0%）、8.7 mM（0.5%）、52 mM（3%）或 87 mM（5%）酒精处理 12 小时。收集上述 HUVEC 细胞的条件培养基，用于培养人骨髓间充质干细胞（BM-MSCs），体外诱导其分化为成骨细胞。酒精饮食会减缓骨骼生长，导致骨质疏松症，损害成骨细胞的骨形成，并通过抑制内皮细胞的增殖和促进其衰老来减少 CD31EMCN H 型血管的形成。与载体相比，酒精处理明显增加了 p16 的表达，同时显著降低了 Bmi-1、CDK6、Cyclin D、E2F1 和骨形态发生蛋白（BMP）2 的表达。酒精通过减少 H 型血管内皮细胞的 BMP2 分泌来抑制 BM-MSCs 向成骨细胞的分化。酒精通过 Bmi-1/p16 信号抑制内皮细胞的增殖和促进其衰老，从而损害 CD31EMCN H 型血管的形成，并通过减少 H 型血管内皮细胞的 BMP2 分泌来抑制 BM-MSCs 向成骨细胞的分化。本研究为开发针对 H 型血管老化内皮细胞的新治疗策略以预防酒精性骨质疏松症提供了依据。
