Yang Junzheng, He Qi, Wang Yunhan, Pan Zhaofeng, Zhang Gangyu, Liang Jianming, Su Lijun, Wang Ailin, Zeng Chuning, Luo Haoran, Liu Lingyun, Li Jianliang, Rao Qiuhong, Wang Baohua, Wang Haibin, Chen Peng
First School of Clinical Medicine, Guangzhou University of Chinese Medicine, 12 Jichang Road, Baiyun District, Guangzhou 510405, PR China; The Laboratory of Orthopaedics and Traumatology of Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, 12 Jichang Road, Baiyun District, Guangzhou 510405, PR China.
Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, 12 Jichang Road, Baiyun District, Guangzhou 510405, PR China.
Phytomedicine. 2022 Jan;94:153810. doi: 10.1016/j.phymed.2021.153810. Epub 2021 Oct 25.
Osteoporosis affects more than half the patients with type 2 diabetes mellitus (T2DM). Up to data, there is no effective clinical practice in managing type 2 diabetes osteoporosis (T2DOP) because of its complex pathogenesis. Gegen Qinlian Decoction (GQD) has been used for the long-term management of T2DM. However, the underlying mechanism of GQD in the treatment of T2DOP remains unknown.
To reveal the role of GQD in T2DOP and its potential therapeutic targets in the management of T2DOP.
The effect of GQD on T2DOP was observed in db/db mice in four groups: model group, GQD low-dose group (GQD-L), GQD high-dose group (GQD-H), and metformin (positive control) group. C57BL/6J mice were used as the negative control group.
Quantitative phytochemical analysis of GQD was performed using high-performance liquid chromatography (HPLC). Micro-CT and hematoxylin-eosin (H&E) staining were used to evaluate bone histomorphometry. To screen for candidate targets of GQD, a cytokine antibody array was used, followed by bioinformatics analysis. Quantitative real-time PCR (qRT-PCR) and western blotting (WB) were used to determine expression levels.
The major active components of GQD were confirmed by HPLC. Micro-CT and H&E staining showed that bone mass was significantly increased in the GQD-H group compared with the model group. Antibody arrays revealed that the expression of insulin-like growth factor binding protein 3 (IGFBP3) was elevated in the GQD-H group. The MAPK pathway was identified using bioinformatics analysis. Additionally, the levels of osteoclastogenesis-related genes, including cathepsin K (Ctsk), acid phosphatase 5 (Acp5), matrix metallopeptidase 9 (Mmp9), and ATPase H+ transporting V0 subunit D2 (Atp6v0d2) were significantly decreased in the GQD-H group. Compared with the model group, high-dosage GQD inhibited phosphorylation of extracellular signal-regulated kinases (ERKs) and P38 mitogen-activated protein kinase (MAPK) and the expression of c-Fos and nuclear factor of activated T cells 1 (NFATc1).
GQD plays a protective role in T2DOP by upregulating IGFBP3 expression and downregulating the IGFBP3/MAPK/NFATc1 signaling pathway. IGFBP3 in serum may also be a novel biomarker in the treatment of T2DOP. Our current findings not only expand the application of GQD, but also provide a theoretical basis and guidance for T2DOP.
骨质疏松症影响超过半数的2型糖尿病（T2DM）患者。截至目前，由于2型糖尿病骨质疏松症（T2DOP）发病机制复杂，尚无有效的临床治疗方法。葛根芩连汤（GQD）已被用于T2DM的长期治疗。然而，GQD治疗T2DOP的潜在机制尚不清楚。
揭示GQD在T2DOP中的作用及其在T2DOP治疗中的潜在靶点。
在db/db小鼠中观察GQD对T2DOP的影响，分为四组：模型组、GQD低剂量组（GQD-L）、GQD高剂量组（GQD-H）和二甲双胍（阳性对照）组。C57BL/6J小鼠作为阴性对照组。
采用高效液相色谱（HPLC）对GQD进行定量植物化学分析。使用微计算机断层扫描（Micro-CT）和苏木精-伊红（H&E）染色评估骨组织形态计量学。为筛选GQD的候选靶点，使用细胞因子抗体芯片，随后进行生物信息学分析。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法（WB）测定表达水平。
通过HPLC确定了GQD的主要活性成分。Micro-CT和H&E染色显示，与模型组相比，GQD-H组骨量显著增加。抗体芯片显示，GQD-H组胰岛素样生长因子结合蛋白3（IGFBP3）表达升高。通过生物信息学分析确定了丝裂原活化蛋白激酶（MAPK）通路。此外，GQD-H组破骨细胞生成相关基因，包括组织蛋白酶K（Ctsk）、酸性磷酸酶5（Acp5）、基质金属蛋白酶9（Mmp9）和ATP酶H+转运V0亚基D2（Atp6v0d2）的水平显著降低。与模型组相比，高剂量GQD抑制细胞外信号调节激酶（ERK）和P38丝裂原活化蛋白激酶（MAPK）的磷酸化以及c-Fos和活化T细胞核因子1（NFATc1）的表达。
GQD通过上调IGFBP3表达和下调IGFBP3/MAPK/NFATc1信号通路在T2DOP中发挥保护作用。血清中的IGFBP3也可能是T2DOP治疗中的一种新型生物标志物。我们目前的研究结果不仅扩展了GQD的应用，也为T2DOP提供了理论基础和指导。
