Research Article: Multi-omics analysis explores the potential mechanisms of Huangtcia renifolia ethanolic extract against osteoporosis in ovariectomized rats
Abstract:
Huangtcia renifolia (L.) H. Ohashi & K. Ohashi, a traditional folk medicine, has been used for centuries by the Dai people in Xishuangbanna, Yunnan Province, China. Traditionally, it has been used to clear kidney heat, promote diuresis, and treat conditions, including kidney deficiency, sexual dysfunction, hepatitis, and osteoporosis. Recent pharmacological investigations indicate that Huangtcia renifolia ethanol extract (DRS) exerts bone-protective effects, partially via modulation of OPG/RANKL/RANK and BMP-2/Smad signaling pathways. This study used an integrated multi-omics strategy to explore the bone-protective effects of DRS and its potential mechanisms in ovariectomized (OVX) rats.
The chemical composition of DRS was profiled by widely targeted UPLC-MS/MS. Osteoporosis was induced in rats via bilateral ovariectomy, and bone status was assessed using serum biochemical analyses, micro-computed tomography (microCT) and hematoxylin-eosin (H&E) staining of femur tissue. Integrated multi-omics analyses were subsequently performed, comprising 16S rRNA sequencing of fecal microbiota, widely targeted serum metabolomics, and femoral transcriptomic sequencing. In vitro validation was performed using MC3T3-E1 osteoblasts treated with DRS-containing serum.
DRS treatment significantly increased bone mineral density, improved trabecular bone microstructure, and effectively reversed abnormal bone turnover markers in OVX rats. Gut microbiota profiling indicated that DRS treatment modulated the gut microbiota, with significant increases in the relative abundance of Lactobacillus and Limosilactobacillus . Serum metabolomics identified 34 metabolites differentially abundant in DRS-treated rats, with pathway enrichment analyses highlighting purine metabolism as one of the principal affected pathways. Femoral transcriptomics and RT-qPCR validation indicated that DRS treatment was associated with upregulation of osteogenic genes ( Col1a1 and Col1a2 ) and downregulation of muscle-related genes ( Myog , Myh3 , and Mstn ), consistent with potential modulation of pathways linking bone and muscle function. Additionally, DRS-containing serum dose-dependently enhanced osteoblast differentiation and upregulated BMP2 and RUNX2 expression in vitro .
DRS treatment protects against osteoporosis in OVX rats. This protection was associated with gut microbiota composition, serum metabolic profiles, and the expression balance between osteogenic and muscle-related genes in bone.
Introduction:
Huangtcia renifolia (L.) H. Ohashi & K. Ohashi, a traditional folk medicine, has been used for centuries by the Dai people in Xishuangbanna, Yunnan Province, China. Traditionally, it has been used to clear kidney heat, promote diuresis, and treat conditions, including kidney deficiency, sexual dysfunction, hepatitis, and osteoporosis. Recent pharmacological investigations indicate that Huangtcia renifolia ethanol extract (DRS) exerts bone-protective effects, partially via modulation of OPG/RANKL/RANK and…
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