Drug and food/drink for preventing or improving cerebral dysfunction
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example 1
[0102]In Example 1, where PC12 cells and RBL2H3 cells were used as aromatic monoamine synthesizing cells and sepiapterin was added to a culture medium, evaluation was made for a total amount of biopterin in the cells (a total amount of tetrahydrobiopterin, and its oxidant, dihydrobiopterin and biopterin, the same shall apply hereinafter).
[0103]The PC12 cells are cultured cells having properties of a neuron and it is known that they synthesize dopamine, noradrenaline and adrenaline, with tetrahydrobiopterin used as a coenzyme. The RBL2H3 cells are cultured cells having properties of a mast cell and it is known that they synthesize serotonin, with tetrahydrobiopterin used as a coenzyme.
[0104]The both cells are capable of synthesizing tetrahydrobiopterin and also retain a certain amount of tetrahydrobiopterin in the cells but do not retain a saturated amount thereof. Furthermore, unlike cells which are not capable of synthesizing aromatic monoamines by themselves but take up aromatic m...
example 2
[0119]In Example 2, a cell system of a brain blood vessel wall model was used to compare passage of sepiapterin with that of tetrahydrobiopterin across a blood vessel wall.
[0120]As the brain blood vessel wall model, there was used a “BBB kit, RBT24H (made by PharmaCo-Cell Company Ltd. in Japan).” This kit was a model system in which rat vascular endothelical cells were cultured on a porous synthetic resin film having small pores of 3 μm in inner diameter to form tight intercellular junctions, thereby forming the blood vessel wall. In this kit, pericytes were cultured in advance on the back side of the porous synthetic resin film and astroglia cells were also cultured at the same time in the well below the film, thereby forming the intercellular tight junctions of vascular endothelical cells. A cultured area was 0.3 cm2 per well. In this model, the cell sheet, an upper side of the cell sheet and a lower side of the cell sheet respectively correspond to the blood vessel wall, an intra...
example 3
[0128]In Example 3, comparison was made between sepiapterin and tetrahydrobiopterin in terms of uptake into astroglia cells.
[0129]The astroglia cells are major glia cells and present tightly around vessels of the brain. This cell selectively takes up a substance which has passed through the blood vessel walls of the brain and supplies the substance to neurons.
[0130]Therefore, CTX TNA2 cells, that is, cultured cells derived from the astroglia cells, were used to evaluate the uptake of sepiapterin and that of tetrahydrobiopterin into the astroglia cells. It is noted that the CTX TNA2 cells used were obtained from the ATCC in the U.S.A. (American Type Culture Collection).
[0131]On a previous day of the experiment, the CTX TNA2 cells were inoculated at 1×105 per well. Thirty minutes after a culture medium was replaced by Hank's-HEPES (pH 7.4), sepiapterin was added at 50 μM or tetrahydrobiopterin was added at 100 μM, each of which was cultured for 0, 5, 10, 20, 40 and 60 minutes. After c...
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