生物化学实验influence of starvation and satiation on liver glycogen

1、Mo H BMedical School, Jinan UninversityInfluence of starvation and satiation on liver glycogen contentBiochemistry Exp Liver glycogen is a macromolecular polyhexose , which is the major form of sugar storage. IntroductionInfluence of starvation and satiation on liver glycogen contentGlycogen Biosynt

2、hesisGluconeogene-sisPentose pathway (Other sugar)Glycogen-olysisFood sugarOxidationFat , amino acidBlood Sugar PoolGlycogen metabolism plays a very important role on regulating the blood sugar! Pure glycogen is easily soluble in water to form an clarity solution, and give a red color with iodine; t

3、he color reaction is indicative, and proportional to,the content of liver glycogen. Glycogen could be extracted with 5% trichloroacetic acid, and determined at 520 nm on a spectrophotometer.Glycogen+I2Glycogen-I2 complex1.Preparation of animals2.Kill the mouse quicklyMethods( Procedure)3. Weigh accu

4、rately 0.2 g of liver tissue4. Put in a 10mL beaker Get 10 ml 5% tricholoracetic by volumetric cylinder Transfer 5ml 5% trichloroacetic to the beakerHomogenate the liver tissue6. wash the tip of homogenizer with left 5 mL 5% tricholoracetic; Stand for 3 min5. Transfer the supernatant to a 10 mL cent

5、rifuge tube7. Centrifuge 10 min, 2000 rpm 8. Transfer the supernatant to a new 10 mL centrifuge tube and determinate the content of liver glucogen.Label 4 tubes and proceed as the following table:Tube no.1 blank2standard3Satiated mouse4Starved mouse5%trichlorsacetic acid2.01.01.01.0Standard glycogen

6、 sol.(0.3mg/ml)-1.0-Extract of glycogen of satiated mouse-1.0-Extract of glycogen of starved mouse-1.0NaCl I2 Reagent3.03.03.03.0mlMix well, and let stand for 10mins. Read the absorbance at 520 nm. The first tube as the blank to adjust the colorimeter to zero absorbance. Determination of glycogenA B

7、 C DCalculation The reading of the label 2 tube is C, the label 3 tube is A, and the label 4 tube is B. To calculate as the following formula: Liver glycogen content of satiation (g%) =A/C0.31/1000 (10+0.2)/1.0 100/0.2 Liver glycogen content of starved (g%) =B/C0.31/1000 (10+0.2)/1.0 100/0.2NOTE1. Label very tube