5'-N-EthylcarboxamIDOadeNOSine 是腺苷受体的激动剂，可增加荧光素和低分子量葡聚糖的脑外渗，与血脑屏障调节无关。

NECA is an agonist of Adenosine receptor, increases cerebral extravasation of fluorescein and low molecular weight dextran independent of blood-brain barrier modulation.

In vitro cultures of an Epo producing hepatocellular carcinoma (Hep3B) cell line with NECA (>or = 10(-6) M) for 20 hours under hypoxic conditions (1% O2) produced significant increases in medium levels of Epo when compared with hypoxia controls. Hepatocellular carcinoma cells treated with NECA at a concentration range of 10(-7) M to 5 x 10(-5) M for one hour in a hypoxic atmosphere also had significantly higher cAMP levels than that of hypoxia controls. Scatchard analyses of [3H]NECA binding to membrane preparations of hepatocellular carcinoma cells showed low affinity binding sites with a dissociation-constant (Kd) of 0.44 microM and a binding capacity of 863 fmol/mg protein. Suggest that the increase in Epo production in response to NECA under hypoxic conditions can be attributed, at least in part, to stimulation of adenosine A2 receptors which is coupled to adenylyl cyclase activation[1].

NECA, an adenosine analogue, on erythropoietin (Epo) production. NECA (0.05 and 0.1 mumol/kg i.v.) produced significant increases in serum Epo levels (368.8 +/- 56.1 and 384.6 +/- 45.9 mU/ml, respectively) in exhypoxic polycythemic mice after a four hour exposure to hypoxia when compared with hypoxia controls (133.2 +/- 18.2 mU/ml). The hypoxic kidney Epo levels were 46.4 +/- 13.4 mU/kg kidney which were significantly higher than normoxic kidney Ep levels (< 1.24 mU/kg kidney). Theophylline (20 mg/kg i.p.), an adenosine receptor antagonist, significantly inhibited the stimulatory effects of NECA on serum Epo levels[1].

A human hepatocellular carcinoma cell line (Hep3B) was employed to determine the in vitro effects of NECA on Epo production and cAMP accumulation. Hep3B cells were carried in a monolayer cell culture and maintained in 75 cm^2 Corning culture flasks containing Eagle's minimal essential medium (MEM) supplemented with 10% fetal bovine serum (PBS), 0.1 m nonessential amino acids, 1 m sodium pyruvate, 100 U/ml penicillin G, and 100 μg/ml streptomycin in a humidified atmosphere of 5% C02/95% air at 37℃. The culture medium was replaced every two days. Cells were detached with trypsin and aliquots of 2.5 x 10^5 viable cells (determined by trypan blue dye exclusion) were transferred to 24 multi-well plates. Each experiment was carried out with low density cells before they reached confluency. The cells were incubated with NECA in concentrations of 1O^9 M to 5 x l0^-5 M in the presence of 1 U/ml adenosine deaminase (ADA) under hypoxic conditions (1% 02, 5% C02, and 94% N2) for 20 hours following 24 hour preincubation in a normoxic atmosphere. At the end of the incubation period, the supernatant was harvested and frozen at -70℃ prior to Epo RIA. The multiple-range test of Duncan was used for the comparison of the several in vitro treatment groups compared with controls[1].

35920-39-9

C12H16N6O4

308.298

5'-N-乙基酰胺基腺苷;NECA

DMSO:150 mg/mL (486.55 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years