Lycium barbarum L.-Derived miR162a Functions on Osteoporosis Through Directly Promoting Osteoblast Formation
a Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing 210022, China
b School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China
c Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Nantong University, Nantong 226001, China
d Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Microbiology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China
e State Administration of Traditional Chinese Medicine Key Laboratory of Chinese Medicinal Resources Recycling Utilization, Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China
f School of Artificial Intelligence and Information Technology, Nanjing University of Chinese Medicine, Nanjing 210023, China
Traditional Chinese medicine (TCM) can help prevent or treat diseases; however, there are few studies on the active substances of TCM. For example, Lycium barbarum L. has been proven to be effective in treating osteoporosis for thousands of years, but its active substance remains to be unknown. Prompted by the efforts to modernize TCM, the present study focused on the novel active substance of Lycium barbarum L. to reinforce kidney essence to produce bone marrow. Illumina deep sequencing analysis and stem-loop polymerase chain reaction (PCR) assay revealed that miR162a, a Lycium barbarum L.-derived microRNA, can pass through the gastrointestinal tract to target the bone marrow in mice. Immunofluorescence staining showed that miR162a was absorbed through systemic RNA interference defective transmembrane family member 1 (SIDT1) in the stomach. Bioinformatics prediction and luciferase reporter assay identified that miR162a targeted nuclear receptor corepressor (NcoR). Alizarin red staining and micro-computed tomography (microCT) confirmed that miR162a promoted osteogenic differentiation in bone marrow mesenchymal stem cells, zebrafish, and a mouse model of osteoporosis. In addition, transgenic Nicotiana benthamiana (N. benthamiana) leaves overexpressing miR162a were developed by agrobacterium infiltration method. microCT and tartrate-resistant acid phosphatase staining confirmed that transgenic N. benthamiana leaves effectively protected against osteoporosis in mice. Our study mechanistically explains how Lycium barbarum L. improves osteoporosis and supports that Lycium barbarum L. reinforces kidney essence, thereby strengthening the bone. miR162a expressed by transgenic plants may represent a novel and safe treatment for human osteoporosis.