BACKGROUND:Chinese nourishing kidney herbs can prevent osteoporosis and improve bone metabolism, which has been proved in animal and cel experiments. But there are few reports on the compatibility of Chinese nourishing kidney herbs, and it is difficult to screen the optimal compatibility, as the interaction of active ingredients and drug substance basis are uncertain. OBJECTIVE: To determine the proliferation, differentiation and Smad4 mRNA expression of neonatal Sprague-Dawley rat osteoblasts cultured by Chinese nourishing kidney herbs with different compatibility so as to find out the optimal compatibility of Chinese nourishing kidney herbs. METHODS: Passage 5 osteoblasts were divided into five groups: group A, 1×10-5mol/L icarin; group B, 1×10-5mol/L icarin+1×10-5 mol/L naringin; group C, 1×10-5mol/L icarin+1×10-5 mol/L diosgenin; group D, 1×10-5mol/L icarin+ 1×10-5mol/L catalpol; group E, 10 μL normal saline (control group). There were six wels in each group. RESULTS AND CONCLUSION: Compared with the group E, the proliferative ability of osteoblasts and expression of Smad4 mRNA were increased in the groups B and C; until the 72nd hour, the proliferative ability of osteoblasts in the group B reached the peak. At 48 hours of culture, the activity of alkaline phosphatase in groups B and C was higher than that in group E; at 72 hours of culture, the activity of alkaline phosphatase in groups B and D was higher than that in group E. These findings indicate that the compatibility of Chinese nourishing kidney herbs can influence the activity of osteoblasts, and icarin+naringin has the strongest effect.

Objective:To study the role of a novel brain-derived peptide hypoxic-ischemic brain damage associated peptide (HIBDAP) in regulating pyroptosis of oxygen-glucose deprived (OGD) microglia.Methods:The sequence of HIBDAP was coupled with the sequence of cell-penetrating peptide transactivator of transcription (TAT) to form TAT-HIBDAP. Fluorescein isothiocyanate (FITC) labeled TAT-HIBDAP was added to microglia cells and observed under fluorescence microscope. Microglia cells were treated with different concentrations of TAT-HIBDAP (1, 5, 10, 20 μmol/L) and then OGD process. Cell pyroptosis was analyzed using lactate dehydrogenase (LDH) assay. The concentration of TAT-HIBDAP with the most prominent inhibiting effects was determined and selected for subsequent experiments. The pyroptosis morphology of the control group, the OGD group and the HIBDAP group (5 μmol/L TAT-HIBDAP+OGD) was observed using transmission electron microscope. The mRNA and protein expression of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasomes were examined using real-time quantitative PCR and Western Blot analysis.Results:Fluorescence microscope showed FITC-labeled TAT-HIBDAP could successfully enter microglia cells. Compared with the OGD group, low concentrations of TAT-HIBDAP (1, 5, 10 μmol/L) could significantly reduce microglia pyroptosis and the concentration of 5 μmol/L showed the most prominent effects. Compared with the control group, OGD group showed typical pyroptosis morphology and HIBDAP group showed significantly improved morphology. The mRNA and protein expression of NLRP3 inflammasomes in the OGD group were significantly higher than the control group and also the HIBDAP group.Conclusions:The novel brain-derived peptide HIBDAP may reduce the expression of NLRP3 inflammasomes and inhibit the pyroptosis of OGD microglia.