Department of Cardiology, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning, China.
Department of Emergency, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning, China.
J Cell Biochem. 2019 Dec;120(12):19660-19672. doi: 10.1002/jcb.29272. Epub 2019 Aug 12.
In this study, we established both an animal model and a cellular model of hyperuricemia (HUC). Subsequently, we treated these models with allopurinol (ALLO) to study the effect of uric acid (UA) and ALLO on the differentiation and proliferation of osteoblasts and vascular smooth muscle cells (VSMC).
Western Blot, immunohistochemistry assay, and real-time polymerase chain reaction were conducted to measure the changes in the expression of differentiation-related factors in osteoblasts and VSMCs in HUC and HUC+ALLO groups. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and flow cytometry were utilized to observe the changes in the proliferation of osteoblasts in HUC and HUC+ALLO groups. Von Kossa staining was performed along with calcium content measurement to investigate the effect of HUC/ALLLO on vascular calcification.
In this study, the levels of Wnt3a and differentiation-related factors, including Runx2, Sp7, Ibsp, Bglap, Dmp1, and Col1a1, were all evidently decreased in HUC rats, while the presence of ALLO increased the levels of above factors. In addition, the viability of osteoblasts was reduced while their apoptosis was elevated in the HUC group, and ALLO treatment reduced the apoptosis and increased the viability of osteoblasts to a certain extent. Moreover, HUC elevated the levels of Wnt3a, Runx2, Sp7, Bglap, Col1a1, SM22a, and Acta2 in VSMCs of HUC rats, leading to greatly increased calcium content and obvious vascular calcification. In contrary, ALLO treatment reduced the effect of HUC. Furthermore, the effect of UA and ALLO on osteoblasts and VSMCs was also validated in cellular models treated with monosodium urate (MSU) crystals or MSU+ALLO.
HUC can suppress the differentiation and proliferation of osteoblasts while promoting the differentiation of VSMCs both in vivo and in vitro. The treatment by ALLO exhibited a therapeutic effect on HUC by promoting the differentiation and proliferation of osteoblasts while reducing vascular calcification.
本研究建立了高尿酸血症（HUC）动物模型和细胞模型，并用别嘌醇（ALLO）处理这些模型，以研究尿酸（UA）和 ALLO 对成骨细胞和血管平滑肌细胞（VSMC）分化和增殖的影响。
采用 Western Blot、免疫组织化学检测和实时聚合酶链反应检测 HUC 和 HUC+ALLO 组中成骨细胞和 VSMC 分化相关因子的表达变化。MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐）法和流式细胞术观察 HUC 和 HUC+ALLO 组中成骨细胞增殖的变化。Von Kossa 染色结合钙含量测定研究 HUC/ALLO 对血管钙化的影响。
本研究中，HUC 大鼠的 Wnt3a 及 Runx2、Sp7、Ibsp、Bglap、Dmp1 和 Col1a1 等分化相关因子的水平均明显降低，而 ALLO 的存在增加了上述因子的水平。此外，HUC 组成骨细胞活力降低，凋亡增加，而 ALLO 处理在一定程度上降低了成骨细胞的凋亡，增加了其活力。此外，HUC 增加了 HUC 大鼠 VSMC 中的 Wnt3a、Runx2、Sp7、Bglap、Col1a1、SM22a 和 Acta2 水平，导致钙含量显著增加和明显的血管钙化。相反，ALLO 处理降低了 HUC 的作用。此外，在单核细胞尿酸盐（MSU）晶体或 MSU+ALLO 处理的细胞模型中，UA 和 ALLO 对成骨细胞和 VSMC 的作用也得到了验证。
HUC 可抑制体内外成骨细胞的分化和增殖，促进 VSMC 的分化。ALLO 治疗通过促进成骨细胞的分化和增殖，减少血管钙化，对 HUC 具有治疗作用。
