二甲双胍减轻糖尿病诱导的大鼠骨质减少与RAGE-JAK2-STAT1信号轴的下调有关。

Metformin attenuates diabetes-induced osteopenia in rats is associated with down-regulation of the RAGE-JAK2-STAT1 signal axis.

作者信息

Lin Rui, Xu Bilian, Ye Zhiqiang, Gao Yin, Fang Haiping, Song Jintong, Liang Dahong, Liu Lingna, Hu Zilong, Zhang Min, Wei Jinsong, Deng Feifu, Zhong Xiangxin, Cui Liao, Liu Yanzhi

机构信息

Zhanjiang Key Laboratory of Orthopaedic Technology and Trauma Treatment, Zhanjiang Central Hospital, Guangdong Medical University, Zhanjiang, 524037, PR China.

Guangdong Provincial Key Laboratory for Research and Development of Natural Drug, School of Pharmacy, Guangdong Medical University, Zhanjiang, 524023, PR China.

出版信息

J Orthop Translat. 2023 Jun 3;40:37-48. doi: 10.1016/j.jot.2023.05.002. eCollection 2023 May.

DOI:10.1016/j.jot.2023.05.002

PMID:37304218

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10250823/

Abstract

BACKGROUND

Osteopenia and fragile fractures are diabetes-associated complications. Many hypoglycemic drugs have effects on bone metabolism. Metformin, as is a prescribed medication for type 2 diabetes mellitus (T2DM), had been reported to have osteoprotective effects beyond its hypoglycemic effect, however the potential mechanism behind these effects remains unclear. In this study, we aimed to investigate the comprehensive effects of metformin on bone metabolism in T2DM rat model and elucidate the potential mechanism.

METHODS

Goto-Kakizaki spontaneous T2DM rats with significant hyperglycemia were treated with/without metformin for 20 weeks. Glucose tolerance was tested and all rats were weighed every two weeks. The osteoprotective effects of metformin in diabetic rats were determined by quantifying serum bone biomarkers, μ-CT imaging, histological staining, bone histomorphometry, and biomechanical properties analyses. Potential targets of metformin in the treatment of T2DM and osteoporosis were predicted using network pharmacology. The effects of metformin on mesenchymal stem cells (C3H10) cultured in high glucose medium were evaluated by CCK-8 assay, alkaline phosphatase (ALP) staining, qPCR and western blotting.

RESULTS

This study demonstrated that metformin significantly attenuated osteopenia, decreased serum glucose and glycated serum protein (GSP) levels, improved bone microarchitecture, and biomechanical properties in GK rats with T2DM. Metformin significantly increased biomarkers of bone formation, and significantly decreased muscle ubiquitin C (Ubc) expression. Network pharmacology analysis found that signal transducer and activator of transcription1 (STAT1) would be a potential target of metformin for regulating bone metabolism. Metformin increased C3H10 cell viability , alleviated ALP inhibition caused by hyperglycemia, increased the osteogenic gene expression of runt-related transcription factor 2 (RUNX2), collagen type I alpha 1 (Col1a1), osteocalcin (OCN), and ALP, while suppressing RAGE and STAT1 expression. Metformin also increased the protein expression of Osterix and decreased that of RAGE, p-JAK2, and p-STAT1.

CONCLUSIONS

Our results demonstrate that metformin attenuated osteopenia and improved bone microarchitecture in GK rats with T2DM and significantly promoted stem cell osteogenic differentiation under high glucose condition. The effects of metformin on bone metabolism are closely associated with the suppression of RAGE-JAK2-STAT1 signaling axis.

THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE

Our research provides experiment evidence and potential mechanistic rationale for the use of metformin as an effective candidate for diabetes-induced osteopenia treatment.

摘要

背景

骨质减少和脆性骨折是糖尿病相关并发症。许多降糖药物对骨代谢有影响。二甲双胍作为2型糖尿病（T2DM）的常用处方药，已被报道除降糖作用外还具有骨保护作用，但其潜在机制仍不清楚。在本研究中，我们旨在探讨二甲双胍对T2DM大鼠模型骨代谢的综合影响，并阐明其潜在机制。

方法

将具有明显高血糖的Goto-Kakizaki自发性T2DM大鼠用/不用二甲双胍治疗20周。每两周测试一次葡萄糖耐量并对所有大鼠称重。通过定量血清骨生物标志物、μ-CT成像、组织学染色、骨组织形态计量学和生物力学性能分析来确定二甲双胍对糖尿病大鼠的骨保护作用。使用网络药理学预测二甲双胍在治疗T2DM和骨质疏松症中的潜在靶点。通过CCK-8测定、碱性磷酸酶（ALP）染色、qPCR和蛋白质印迹法评估二甲双胍对在高糖培养基中培养的间充质干细胞（C3H10）的影响。

结果

本研究表明，二甲双胍显著减轻了T2DM GK大鼠的骨质减少，降低了血糖和糖化血清蛋白（GSP）水平，改善了骨微结构和生物力学性能。二甲双胍显著增加了骨形成生物标志物，并显著降低了肌肉泛素C（Ubc）表达。网络药理学分析发现，信号转导和转录激活因子1（STAT1）可能是二甲双胍调节骨代谢的潜在靶点。二甲双胍增加了C3H10细胞活力，减轻了高血糖引起的ALP抑制，增加了 runt相关转录因子2（RUNX2）、I型胶原α1（Col1a1）、骨钙素（OCN）和成骨细胞碱性磷酸酶（ALP）的成骨基因表达，同时抑制了晚期糖基化终末产物受体（RAGE）和STAT1表达。二甲双胍还增加了osterix的蛋白表达，降低了RAGE、p-JAK2和p-STAT1的蛋白表达。