Biochemical characterization of an ectonucleotide pyrophosphatase/phosphodiesterase (E-NPP, E.C. 3.1.4.1) from rat cardiac soluble and microsomal fractions

Pochmann, Daniela; Innocente, Adrine Maria; Buffon, Andréia; Sarkis, João José Freitas; Porciúncula, Lisiane de Oliveira

doi:10.3109/14756366.2011.574129

Cited by 2 publications

(1 citation statement)

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“…The activity of NPP1 was determined at day 3 and 5 of mineralization by quantifying the hydrolysis of p-nitrophenyl 5 -thymidine monophosphate (p-Nph-5 -TMP) and the resulting change in color [46]. Cells were rinsed twice with 50 mM Tris-HCl (pH 7.35) then scraped in 500 µL of 0.05% Triton-X100 Tris-HCl and transferred to 1.5-mL Eppendorf tubes.…”

Section: Npp1 Activity Assaymentioning

confidence: 99%

Red Rooibos Tea Stimulates Osteoblast Mineralization in a Dose-Dependent Manner

et al. 2019

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Most studies show a positive association between higher tea intake and bone mineral density (BMD), while associations with fracture are inconclusive. Although a few studies have investigated the potential effects of rooibos tea on the bone cell metabolism, only 1 or 2 doses were tested or specific flavonoids were evaluated in isolation. A dose-response study is needed to comprehensively identify whether a level that modulates bone cell activity is associated with dietary or supplemental levels of red rooibos (RR) tea. We investigated if RR tea stimulates mineralization in a dose-dependent manner. Mineralization was induced in Saos-2 cells in the presence of RR tea (0.1–10 µg/mL of polyphenols) or control (dH2O) for 5 days. Cell activity, mRNA, protein expression and activity of key regulators of mineralization were measured. A positive dose-dependent relationship was observed between RR tea concentration and mineralization accompanied with similar trends in cell activity. With high concentrations of tea, protein expression of enzymes directly regulating mineralization (ALP and NPP1) were unchanged but ALP activity was significantly elevated. Osteopontin (OPN) was significantly downregulated midway through mineralization while sclerostin (SOST) was upregulated in response to higher concentrations of RR tea. In conclusion, RR tea stimulated osteoblast mineralization in a dose-dependent manner and were accompanied with downregulation of OPN, increased ALP activity, and increased cell activity.

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Section: Npp1 Activity Assaymentioning

confidence: 99%