Wen-Shen-Tong-Luo-Zhi-Tong Decoction regulates bone–fat balance in osteoporosis by adipocyte-derived exosomes

Abstract

Context

Wen-Shen-Tong-Luo-Zhi-Tong (WSTLZT) Decoction is a Chinese prescription with antiosteoporosis effects, especially in patients with abnormal lipid metabolism.

Objective

To explore the effect and mechanism of WSTLZT on osteoporosis (OP) through adipocyte-derived exosomes.

Materials and methods

Adipocyte-derived exosomes with or without WSTLZT treated were identified by transmission electron microscopy, nanoparticle tracking analysis (NTA) and western blotting (WB). Co-culture experiments for bone marrow mesenchymal stem cells (BMSCs) and exosomes were performed to examine the uptake and effect of exosome in osteogenesis and adipogenic differentiation of BMSC. MicroRNA profiles, luciferase and IP were used for exploring specific mechanisms of exosome on BMSC. In vivo, 80 Balb/c mice were randomly divided into four groups: Sham, Ovx, Exo (30 μg exosomes), Exo-WSTLZT (30 μg WSTLZT-exosomes), tail vein injection every week. After 12 weeks, the bone microstructure and marrow fat distribution were analysed by micro-CT.

Results

ALP, Alizarin red and Oil red staining showed that WSTLZT-induced exosomes from adipocyte can regulate osteoblastic and adipogenic differentiation of BMSC. MicroRNA profiles observed that WSTLZT treatment resulted in 87 differentially expressed miRNAs (p < 0.05). MiR-122-5p with the greatest difference was screened by q-PCR (p < 0.01). The target relationship between miR-122-5p and SPRY2 was tested by luciferase and IP. MiR-122-5p negatively regulated SPRY2 and elevated the activity of MAPK signalling pathway, thereby regulating the osteoblastic and adipogenic differentiation of BMSC. In vivo, exosomes can not only improve bone microarchitecture but also significantly reduce accumulation of bone marrow adipose.

Conclusions

WSTLZT can exert anti-OP effect through SPRY2 via the MAKP signalling by miR-122-5p carried by adipocyte-derived exosomes.

Keywords:

• Bone marrow mesenchymal stem cell
• miR122-5p
• SPRY2
• osteoblastic and adipogenic differentiation
• MAKP signalling

Acknowledgements

We thank all the staff of Lab of Professor Tonghui Ma in Nanjing University of Chinese Medicine.

Author contributions

LW and YG contributed to conception and design of study. JW, ZH and JS contributed to acquisition of data. LY, ML and ZY contributed to analysis of data. LW and YP contributed to drafting the manuscript. YM, ML and PT contributed to revising the manuscript critically for important intellectual content. All authors approved the version of the manuscript to be published.

Disclosure statement

The authors declare no conflict of interest.

Data availability statement

The data sets used and/or analysed during the current study or supplementary material are available from the corresponding author on reasonable request.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (82074458, 82174411). This research is also sponsored by Natural Science Foundation of Jiangsu Province (BK20221351, BK20220470), Natural Science Foundation of the Jiangsu Higher Education Institution of China (22KJB360012) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (Integration of Chinese and Western Medicine).