Abstract
First documented in Shennong Bencao Jing (about 200 B.C.–200 A.D.), Elaphuri Davidiani Cornu (EDC) has been recorded for its effects in strengthening bones and balancing other aspects of overall health for approximately 2000 years. In the present study, our aim was to investigate which are the components of the active EDC fraction by a peptidomic strategy. We explored the extent to which EDC increases the proliferation of osteoblasts by measuring the elevations in osteonectin and type I collagen mRNA levels and characterized it using nano-flow liquid chromatography in tandem with orbitrap mass spectrometry. In total, 272 peptide sequences from collagens were determined. “Hot regions” in parent proteins determined by peptide heat maps which indicated that amino acid sequences in the regions might undergo proteolysis easily and generate peptides. Among the identified peptides, 90.2% were hydrophilic, and the molecular weight of 97.1% of identified peptides was lower than 2000 Da. According to these results, EDC collagen-derived peptides were easily analyzed and identified. Moreover, this methodology is feasible to characterize the active peptides matrices originated from collagen hydrolysates or some other animal horn- derived TCMs.
Highlights
Animal horns are pointed projections on the head of animals, and are an important part of traditional Chinese medicines (TCMs)
The methylthiazole tetrazolium (MTT) method was used to observe the effect of ECDF on the proliferation of osteoblasts
The EDCF-induced osteoblast differentiation was further confirmed by activation of osteonectin and type I collagen (COL1A1)
Summary
Animal horns are pointed projections on the head of animals, and are an important part of traditional Chinese medicines (TCMs). Most of the Milu deer in the Nanyuan Royal Hunting Garden of the Qing dynasty were killed, to be consumed by starving. Molecules 2017, 22, 166 peasants in 1895 after the hunting garden was destroyed by a flood on the Yongding River. 1900, the remaining Milu deer were shot and eaten by troops during the Boxer Rebellion, making Milu deer extinct in China [1]. As a species under first rank state protection in China, Milu deer are well protected. Since the reintroduction of Milu deer from the United Kingdom, the population has significantly increased from 18 to over 3000 in the last three decades in China [2]. The antlers of Milu deer are naturally shed every year, and can be collected, as easy obtained components, there were sufficient Milu deer antler resources for our scientific study
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