To investigate the effects of different concentrations of Yinyanghuo (Epimedii Folium) extract on osteogenic differentiation of the rats adipose-derived stem cells (ADSCs).

Five healthy, nulliparous SPF-grade female SD rats were used to isolate and culture the ADSCs. The obtained third-generation ADSCs were stained with Alizarin Red and Oil Red O to evaluate their osteogenic and adipogenic differentiation abilities after being cultured with 1, 10, 50, 100 μmol/L Yinyanghuo (Epimedii Folium) extract. Then, the third-generation ADSCs were divided into a blank group, a resveratrol group (100 μmol/L) and Yinyanghuo (Epimedii Folium) extract groups. According to the different concentrations of Yinyanghuo (Epimedii Folium) extract, four subgroups were created, namely Ⅰ～Ⅳ groups (1, 10, 50, 100 μmol/L). After 7 days of culture, qRT-PCR and Western blotting were performed to detect the expression levels of LIM domain-containing protein Ajuba (Ajuba), Runx-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) in the ADSCs.

The resveratrol group and Yinyanghuo (Epimedii Folium) extract I to IV groups all upregulated the expression levels of Runx2 gene and protein, as well as ALP gene, with the best effect observed in the 100 μmol/L Yinyanghuo (Epimedii Folium) extract group (P<0.01). As for the expression levels of ALP protein, the resveratrol group exhibited a stronger promoting effect than the Yinyanghuo (Epimedii Folium) extract I to IV groups (P<0.01). The resveratrol group and Yinyanghuo (Epimedii Folium) extract groups I to IV all downregulated the expression levels of Ajuba gene and protein (P<0.01, P<0.05), with the resveratrol group showing a stronger gene inhibitory effect at the same concentration.

Both Yinyanghuo (Epimedii Folium) extract and resveratrol can promote the osteogenic differentiation of adipose-derived stem cells, but the promoting effect of Yinyanghuo (Epimedii Folium) extract is more significant and effective. Both can also inhibit adipogenic differentiation, with Yinyanghuo (Epimedii Folium) extract exhibiting weaker inhibition than resveratrol in the short term. With the extension of intervention time, further research is needed to investigate the trend in the future.