The Regulative Effection Of NEFA,Ins,HGF,IGF-1 On Hepatocyte InsR Of Calf

Ketosis and fatty liver based on energy metabolism disorder is a nutrition metabolic disease related to hepatic lipid metabolism and develop in periparturient dairy cows generally. Insulin receptor like other protein hormone receptors is embedded in the plasma membrane. Insulin receptor is a tyrosine kinase that is composed of two alpha subunits and two beta subunits linked by disulfide bonds. Insulin receptor plays an important role in various physiological functions such as mediating both IGF-2 and insulin action, further mediate metabolic responses. Therefore, primary hepatocyte culture in vitro, and to add to Nonesterified fatty acids, insulin, hyperglycemic-glycogenolytic factor, insulin-like growth factor-1 with different level in this ,then extract membrane protein and study the activity of hepatocytes insulin receptor.This experiement investigate the regulative effection of metabolic product and neuroendocrine factor on hepatocyte insr of newborn calf and also provide experiment and theory evidence for prevention and cure ketosis and FL , perinatal period energy metabolism disorder.First of all, hepatocyte of newborn calf was adopt perfusion culture metho to make primary culture. The result displays: after 24 h'cultivation, the concentration of hepatocyte reaches the standard of the extraction of membrane protein. That is 3-5*10 cell per cultivate hole. We put NEFA,IRHGF,IGF-1 to invetigate hepatocyte INSR activity fluntuation law. Concentration of the additives was as NEFA: 0mmol/L, 0.5mmol/L, 1.0mmol/L, 1.5mmol/L, 2.0mmol/L,;Ins:0umol/L, 2.0umol/L, 10.0umol/L, 20.0umol/L, 30.0umol/L,100.0umol/L;HGF:0pg/ml,50pg/ml, 100pg/ml, 200 pg/ml, 500 pg/ml,800 pg/ml;IGF-1:0 ng/ml,10 ng/ml,30 ng/ml,50 ng/ml,100 ng/ml,150 ng/ml。 After 10 h'concentration,we abstract membrane protein of hepatocyte and test the activity of hepatocy insr by two point combination test method. The result displays :with the increasing of NEFA's concentration, the contract between the Cogroup and the blank group is significant. NEFA has the down regulation to the activity of hepatocy insr. When the concentration of Insr are 2and 10 umol/L, the activity of hepatocy Insr is lower than CG. When the concentration of Insr is more than 10 umol/L, the activity of hepatocy Insr advance gradually. When the concentration was up to 20 umol/L, the activity of hepatocy Insradvanced gradually, but still lower than CG.When the concentration of 1GF-1 are 10 and 30ng/nl,the activity of hepatocyte Insr is lower than CG,the contract is obviously ,but contract between the two group is not obviously. When the concentration of IGF-1 is more than 30ng/ml, the activity of hepatocyte Insr advance gradually; when the concentration of 1GF-1 is more than 50ng/ml; the concentration of IGF-1 is 50na/ml; the activity of hepatocyte Insr decrease gradually; when the concentration of 1GF-1 is 150ng/ml, the activity of hepatocyte Insr decreased to the CG level. Low concentration 1GF-1 has down regulation effect on OBB primary culture of the activity of hepatocyte Insr equal or higher than physiology level. 1GF-1 has up-rugulation effect on the activity of hepatocyte Insr.but with the heghten of the concentration of IGF-1, the up-regulation to the activity of hepatocyte InsR has descendened .above consequence shows, NEFA has the inhibitory action on OBB primary culture of the activity of hepactocyte InsR, aggravate energy negatively balance,intensify energy negtive balance; low concentration Ins can depress the activity of hepatocyte InsR, maybe because of fat mobilization mechanism;HGF can increase the activity of hepatocyte InsR,when the energy negtive balance occurs ,we can let the calf take or vein shot quantitative HGF which can inhibit fat mobilization, decrease ketone body's production and the deposit of TG. Low concentration IGF-1 can decrease the activity of hepactocyte InsR.when IGF-1 is equal to or more than the physio-level, it can increase the activity of hepatocyte InsR, so quantitative IGF-1 can increse InsR to ins'saitivity.,and has cooperte function to Ins .