Cognitive function improving agent
A technology of cognitive function and improving agent, which can be used in testing pharmaceutical preparations, plant raw materials, antibodies, etc., and can solve problems such as ignorance
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manufacture example 1
[0107] Production Example 1 Production of anti-active GIP antibody
[0108] (1) Synthesis of peptides for immunization
[0109] After adding polyethylene glycol (PEG (polyethyleneglycol)) to the N-terminal 15 amino acids (GIP (1-15)) of active GIP, chemically binds keyhole limpet hemocyanin (KLH; keyhole limpet hemocyanin) , making KLH combined with PEGylated GIP (1-15) to form an immune antigen. A peptide obtained by PEGylation of the N-terminal 15 amino acids of active GIP (GIP(1-15)) was used as the assay antigen (1), and a peptide obtained by PEGylation of the N-terminal 13 amino acids of inactive GIP (GIP(3- 15)) The peptide obtained by PEGylation was used as the antigen for measurement (2).
[0110] (2) immunity
[0111] Immunization was carried out subcutaneously on the back using BALB / c mice (Oriental Yeast Co., Ltd.). In the primary immunization, an emulsion obtained by mixing the above-prepared antigen and complete Freund's adjuvant was administered. From the fi...
Embodiment 2
[0140] Example 2 The age-related change of GIP amount in blood
[0141] (1) Animals and feeding methods
[0142] After introducing C57BL / 6J male mice (Japan Clea Co., Ltd.) at the age of 4 weeks (room temperature 23°C, humidity 55±10%, diurnal rhythm; 7:00~19:00), under free access to food and water for feeding. The diet was CE-2 (Japan Clea Co., Ltd.), and after 1 week of acclimation, they were fed with normal diet (D12450K, Research Diets, Inc) or high-fat diet (D12451, Research Diets, Inc) for 95 weeks.
[0143] (2) blood collection
[0144] Whole blood was collected from the abdominal veins of mice of each week (5, 10, 15, 20, 30, 40, 50, 65, 80, and 100 weeks) under isoflurane anesthesia.
[0145] (3) Determination of GIP amount in blood
[0146] Regarding the concentration of GIP in blood, the collected blood was centrifuged to prepare a plasma fraction, and then the total GIP was measured by a conventional method using a GIPELISA kit (manufactured by Millipore).
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Embodiment 3
[0151] Example 3 Anti-GIP Antibody Improving Effects on Memory and Learning Functions of Aged Mice
[0152] (1) Animals and feeding methods
[0153] After introducing C57BL / 6J male mice (Japan Clea (strain)) at the age of 4 weeks (room temperature 23 ° C, humidity 55 ± 10%, diurnal rhythm; 7:00 ~ 19:00), free to eat (D12450K, Research Diets, Inc) and free drinking water were raised for 111 weeks.
[0154] (2) Preparation of anti-GIP antibody solution
[0155] The anti-active GIP antibody produced in Production Example 1 was dissolved in physiological saline at a concentration of 0.05 mg / mL to form an anti-GIP antibody solution.
[0156] (3) Dosage and method of administration
[0157] C57BL / 6J mice (107 weeks old) were intraperitoneally administered physiological saline (control group) or anti-GIP antibody solution (0.5 mg / kg body weight) once a week (AM9:00-10:00) for a total of 6 times. ) (anti-GIP antibody administration group).
[0158] (4) Y word maze (short-term mem...
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