Wei Yuchi, Wang Mingyue, Jiang Zhanliang, Jia Yuyan, Si Yongmei, Peng Zeyu, Yang Jie, Shi Ye, Wu Yongji, Ding Xiaolei, Pan Daian, Zhao Daqing, Leng Xiangyang, Li Xiangyan, Dong Haisi
Affiliated Hospital of Changchun University of Traditional Chinese Medicine, Changchun University of Chinese Medicine, 130117, Changchun, Jilin Province, China.
College of Traditional Chinese Medicine, Changchun University of Chinese Medicine, 130117, Changchun, Jilin Province, China.
J Ethnopharmacol. 2025 Feb 11;341:119346. doi: 10.1016/j.jep.2025.119346. Epub 2025 Jan 10.
Type 2 diabetic osteoporosis (T2DOP) is a metabolic bone disease characterized by impaired bone structure and decreased bone strength in diabetic patients. Jiangu Decoction (JGD), a traditional Chinese poly-herbal formulation, has shown efficacy in mitigating osteoporosis (OP) and fractures caused by osteoporosis in diabetic patients in clinical trials. In addition, JGD has been proven to promote the proliferation of osteoblasts. However, the specific mechanisms underlying these effects remain unclear.
This study aimed to elucidate the molecular mechanisms underlying the therapeutic effects of JGD in treating T2DOP.
Liquid chromatography-mass spectrometry (LC-MS) was utilized to elucidate the chemical profile of JGD. A T2DOP mouse model (C57BL/6) was established by combining a high-fat diet with streptozotocin (STZ). Micro-computed tomography (micro-CT) imaging, three-point bending tests, and histological staining were utilized to assess alterations in bone mass, bone quality, and bone strength in mice. Mouse Calvaria 3T3-E1 (MC3T3-E1) cells were treated with 33 mmol/L D-glucose (HG), and the protective effect of JGD on the high glucose injury model was observed. Western blotting and qRT-PCR were employed to analyze alterations in biomarkers associated with the Keap1/Nrf2/HO-1 signaling pathway, both in vivo and in vitro.
A total of 909 compounds were identified in JGD using LC-MS. Subsequently, the function of JGD was evaluated both in vitro and in vivo. The findings indicated that JGD promoted bone formation, enhanced bone microstructure, and ameliorated diabetic symptoms in T2DOP mice. Additionally, JGD increased alkaline phosphatase (ALP) activity, facilitated bone mineralization, and upregulated the expression levels of osteogenic marker genes such as runt-related transcription factor 2 (Runx2), osteocalcin (Ocn), and collagen type 1 alpha (Col1a1). Importantly, JGD reduced oxidative stress levels and decreased the accumulation of reactive oxygen species by modulating the Keap1/Nrf2/HO-1 axis both in vivo and in vitro.
Our study suggests that JGD could alleviate T2DOP impairment, closely linked to the Keap1/Nrf2/HO-1 signaling pathway.
2型糖尿病性骨质疏松症（T2DOP）是一种代谢性骨病，其特征是糖尿病患者的骨结构受损和骨强度降低。健骨汤（JGD）是一种传统的中药复方制剂，在临床试验中已显示出对糖尿病患者骨质疏松症（OP）及骨质疏松性骨折具有缓解作用。此外，JGD已被证明能促进成骨细胞的增殖。然而，这些作用的具体机制仍不清楚。
本研究旨在阐明JGD治疗T2DOP的分子机制。
采用液相色谱-质谱联用（LC-MS）技术阐明JGD的化学特征。通过高脂饮食联合链脲佐菌素（STZ）建立T2DOP小鼠模型（C57BL/6）。利用微计算机断层扫描（micro-CT）成像、三点弯曲试验和组织学染色评估小鼠骨量、骨质量和骨强度的变化。用33 mmol/L D-葡萄糖（HG）处理小鼠颅骨3T3-E1（MC3T3-E1）细胞，观察JGD对高糖损伤模型的保护作用。采用蛋白质免疫印迹法（Western blotting）和实时定量聚合酶链反应（qRT-PCR）分析体内外与Keap1/Nrf2/HO-1信号通路相关生物标志物的变化。
利用LC-MS在JGD中鉴定出共909种化合物。随后，在体外和体内对JGD的功能进行了评估。结果表明，JGD促进了T2DOP小鼠的骨形成，增强了骨微结构，并改善了糖尿病症状。此外，JGD增加了碱性磷酸酶（ALP）活性，促进了骨矿化，并上调了成骨标记基因如 runt相关转录因子2（Runx2）、骨钙素（Ocn）和I型胶原α1（Col1a1）的表达水平。重要的是，JGD在体内和体外均通过调节Keap1/Nrf2/HO-1轴降低了氧化应激水平，减少了活性氧的积累。
我们的研究表明，JGD可以减轻与Keap1/Nrf2/HO-1信号通路密切相关的T2DOP损伤。
