本研究旨在探究苦参酮（ORI）对雌激素剥夺所致小鼠骨质疏松的影响及其机制。本研究采用动物实验验证了ORI的抗骨质疏松功效，经微CT进行形态学分析，使用特殊的蛋白质仪检测了小鼠血清中的免疫球蛋白lgM、lgG、补体C3和C4的含量。采用流式细胞术检测小鼠CD4+CD25+Foxp3+调节性T（Treg）细胞和CD4+/CD8+淋巴细胞亚群的表达，采用酶联免疫吸附法检测胰岛素样生长因子（IGF-1）、肿瘤坏死因子（TNF-α）、白细胞介素-1（IL-1）和白细胞介素-6（IL-6）的含量。此外，使用Western blotting检测了Wnt3a/β-连环蛋白信号通路中的关键信号分子。结果显示，与模型组相比，ORI组小鼠股骨中的钙和磷含量增加，脾指数下降。高、中剂量ORI组小鼠血清中碱性磷酸酶（ALP）活性下降，子宫指数增加。ORI显著增加了小鼠股骨的最大弯曲负载和最大弯曲应力，增加了小梁的数量，修复了骨骼的微观结构。同时，ORI可以显著提高免疫球蛋白（lgG和IgM）和补体（C3和C4）的水平，提高小鼠腹膜巨噬细胞的活性，增加脾脏中CD4+CD25+Foxp3+ Tregs和CD4+/CD8+的表达，增加IGF-1的含量，降低TNF-α、IL-1和IL-6的含量，增加股骨组织中VEGF、Wnt3a、p-GSK3β/GSK3β和β-连环蛋白/Lamin的表达水平。这些结果表明，ORI可能通过Wnt3a/β-连环蛋白信号通路调节VEGF的表达，提高小鼠的免疫力、维持免疫系统平衡、促进血管生成，从而改善小鼠的骨密度和骨组织形态，起到抗骨质疏松作用。 版权所有 ©2023 Elsevier B.V.

This study was performed with the aim of investigating the effect of oridonin (ORI) on estrogen deprivation-induced osteoporosis in mice and its mechanism. Animal experiments were used in this work to validate the anti-osteoporotic efficacy of ORI. Morphometric analysis was performed by micro-CT. A special protein meter was used to detect the content of immunoglobulin lgM, immunoglobulin lgG, complement C3 and C4 in the serum of mice. The expression of CD4+CD25+Foxp3+ Treg cell and CD4+/CD8+ lymphocyte subsets in mice was detected by flow cytometry. ELISA was used to detect the content of insulin-like growth factor (IGF-1), tumor necrosis factor (TNF-α), interleukin-1 (IL-1) and interleukin-6 (IL-6). In addition, key signaling molecules in the Wnt3a/β-catenin signaling pathway were detected by Western blotting. The results showed that compared with the model group, the contents of calcium and phosphorus in the femurs of mice in the ORI groups were increased, and the spleen coefficient was decreased. The ALP activity in the serum of mice in the high and medium dose ORI groups was decreased, and the uterine coefficient was increased. ORI significantly increased the maximum bending load and the maximum bending stress of the femurs of mice, increased the number of trabeculae, and repaired the bone microstructure. At the same time, ORI could significantly increase the levels of immunoglobulin (lgG and lgM) and complement (C3 and C4), increase the activity of peritoneal macrophages in mice, increase the expression of CD4+CD25+Foxp3+ Tregs and CD4+/CD8+ in the spleen, increase the content of IGF-1, reduce the content of TNF-α, IL-1 and IL-6 and increase the expression levels of VEGF, Wnt3a, p-GSK3β/GSK3β and β-catenin/Lamin in the femoral tissue. These results indicated that ORI might regulate the expression of VEGF through the Wnt3a/β-catenin signaling pathway, improve the immunity of mice, maintain the balance of the immune system, and promote angiogenesis, thereby improving the bone mineral density and bone tissue morphology of mice and playing an anti-osteoporotic role.Copyright © 2023 Elsevier B.V. All rights reserved.