Liuwei Dihuang pills attenuate ovariectomy-induced bone loss by alleviating bone marrow mesenchymal stem cell (BMSC) senescence via the Yes-associated protein (YAP)-autophagy axis

Abstract

Context

Liuwei Dihuang pill (LWDH) has been used to treat postmenopausal osteoporosis (PMOP).

Objective

To explore the effects and mechanisms of action of LWDH in PMOP.

Materials and methods

Forty-eight female Sprague-Dawley rats were divided into four groups: sham-operated (SHAM), ovariectomized (OVX), LWDH high dose (LWDH-H, 1.6 g/kg/d) and LWDH low dose (LWDH-L, 0.8 g/kg/d); the doses were administered after ovariectomy via gavage for eight weeks. After eight weeks, the bone microarchitecture was evaluated. The effect of LWDH on the differentiation of bone marrow mesenchymal stem cells (BMSCs) was assessed via osteogenesis- and lipogenesis-induced BMSC differentiation. The senescence-related biological indices were also detected using senescence staining, cell cycle analysis, quantitative real-time polymerase chain reaction and western blotting. Finally, the expression levels of autophagy-related proteins and Yes-associated protein (YAP) were evaluated.

Results

LWDH-L and LWDH-H significantly modified OVX-induced bone loss. LWDH promoted osteogenesis and inhibited adipogenesis in OVX-BMSCs. Additionally, LWDH decreased the positive ratio of senescence OVX-BMSCs and improved cell viability, cell cycle, and the mRNA and protein levels of p53 and p21. LWDH upregulated the expression of autophagy-related proteins, LC3, Beclin1 and YAP, in OVX-BMSCs and downregulated the expression of p62.

Discussion and conclusions

LWDH improves osteoporosis by delaying the BMSC senescence through the YAP-autophagy axis.

Keywords:

• Traditional Chinese medicine
• postmenopausal osteoporosis (P MOP)
• senescence

Acknowledgements

We are grateful for the technical support provided by the State Key Laboratory of Southwestern Chinese Medicine Resources (Pharmacy School, Chengdu University of Traditional Chinese Medicine, Chengdu 611130, China).

Author contributions

Bing Liang, Xiongbin Chen and Min Li conceived the study, finished doing all the experiments, compiled and analysed the data, and wrote the manuscript. Xia Yang provides language support for the manuscript. Lingling Zhang, Liangqin Shi, Yanju Gong, Yuanyuan Gong and Huan Xu participated in the design of this study, performed data collection and analysis, manuscript preparation and revision. Xiao Wu, Zhong Jin, Yanru Wang and Luwei Liu contributed to animal experiments, data collection, analysis and manuscript preparation. Xiaohong Yi, Lushuang Xie, Hua Zhong, Chongyang Shen and Yong Wang assisted in the cell experiments and provided guidance on experimental methods and techniques. Lan Yang designed and conceived the study, critically revised the manuscript and finally reviewed it. All authors reviewed the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data that support the findings of this study are available from the corresponding author Lan Yang upon reasonable request.

Additional information

Funding

This study was supported by grants from the Scientific Research Foundation of Chengdu University of Traditional Chinese Medicine (Grant no. JSZX2018004, XCZX2022001), Xinglin Scholar Research Promotion Project of Chengdu University of TCM (Grant no. QNXZ2020002), Scientific Research Start-up Fund (2021) for Introduced Talents (Grant no. 030041251/030) and The National Natural Science Foundation of China (Grant no. 82174226), Open Project of Sichuan Provincial Key Laboratory for Human Disease Gene Study (Grant no. 2023kflx003).