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Objective: To explore the synthesis hexamethyl myricetin (hexamethylmyricetin, HMM) method, the detection method. : Myricetin (myricetin, Myr) as raw material, through methylating agent dimethyl sulfate (Me 2 SO 4 ) reaction, methylation structure modification. HMM-high performance liquid detection method. Orthogonal test method, the proportion of the reactants in the synthesis process, the reaction temperature, to optimize the reaction time of three factors. Sampling at different time points after the reaction starts, and HPLC detection product ultraviolet absorption, to determine the degree of progress of the reaction. The solvent was removed and washed with water to obtain a crude product, after anhydrous recrystallized from ethanol, filtered harvest endproducts, and the use of instruments such as NMR, mass spectrometry, analysis of the structure of the product, and the structure confirmation. Results: ① The final product was MS H NMR 13 the C NMR conclusive evidence of its structure as hexamethyl Myricetin; ② The end products hexamethyl Bayberry factors yield 53 %, purity of greater than 99.84% (HPLC peak area ratio); ③ The optimized reaction conditions were: the proportion of reactants the equivalence ratio 1:7:9 (MYR: Me 2 SO 4 : K 2 the CO 3 ); reaction temperature of 50 ° C; reaction time of 9 hours. Conclusion: The synthetic route is simple, operability, less pollution, high synthesis yield and purity, and low cost. Of purpose of the second part hexamethyl Myricetin hKv1.5 channel: study hexamethyl Myricetin hKv1.5 channel. Method: transfection and stable expression the hKv1.5 (IKur current) channel HEK293 cells, whole-cell patch clamp technique, record of drugs on IKur current in the voltage clamp mode. Results: The positive control of 10 μm drug verapamil can significantly inhibit IKur, its inhibition rate was 87 ± 5%, can be partially restored after elution, the current is recorded by the prompt IKur. Given 0.1,0.3,1,3 μM of the HMM, the current value before the administration of 94.1 ± 3.0% (n = 5), 88.4 ± 5.0% (n = 5), 69.8 ± 4.0% (n = 5), 33.6 ± 2.0% (n = 4). By fitting calculation, HMM inhibition IC50 of 1.79μM IKur of. Conclusion: HMM strong inhibition of hKv1.5 channel. The third part of the domestic FGF-21 KKAy and ob / ob mice purposes: To study the role of domestic FGF-21 KKAy and ob / ob mice. Method: ① KKAy mice, feeding cage, given high fat diet; ob / ob mice, 5 per cage rearing, given a normal diet; controlled room temperature 23 ± 2 ℃, 50% humidity, 12h light-dark cycle (7:00-19: 00), freedom of the consumption of drinking water. ② ophthalmic scissors KKAy ob / ob mice toes cut a small mouth, whole blood oozing sufficient quantities (approximately 0.6μl instrument minimum sample volume requirements), using a Roche blood glucose meter testing blood glucose record blood glucose and blood collection time. ③ oral glucose tolerance test (OGTT, oral glucose tolerance test). Mice were fasted 2h 0 min measured blood sugar, oral administration of glucose solution of 2 g / kg, respectively, given glucose solution after 30min, 60min, 120min testing blood glucose. Results: ① KKAy mice tested samples FGF-21 is not sensitive; fasting glucose reduction ② of FGF-21 low-dose (0.6 mg / kg / d) of ob / ob mice, but no significant difference; FGF -21 high dose (2.5 mg / kg / d) to reduce the role of the ob / ob mice postprandial glucose administered continuously for 7 days, the hypoglycemic effect significant difference; ③ OGTT results show that FGF-21 low-dose (0.6 mg / kg / d) of ob / ob mice with modest improvement of impaired glucose tolerance. Conclusion: domestic FGF-21 has a certain anti-diabetic effects, mainly to reduce postprandial blood glucose, improve impaired glucose tolerance. Compared with the literature, domestic FGF-21 titers Lilly and Company, 1/10, recommended that manufacturers improve the process.
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