Protective Effects Of Reduced Glutathione And Polyene Phosphatidylcholine On Primary Cultured Human Hepatocytes

Objective:The study of the protective effects of the prototyping glutathione and polyenophospholide choline on the cultivation of the liver cells of the adult population after separation.Methods:Choose adult specimens of benign lesions of liver resection,separation and purification using the improved collagenase perfusion method of liver cells,were observed under inverted microscope,blood count method count,preliminary inspection with trypan blue from dying cell activity.Experiment is divided into two parts,the first part:according to the reduced glutathione and polyene lecithin choline concentration is different,divided into GSH 0.1 mmol/L?1 mmol/L?2mmol/L?5 mmol/L?10 mmol/Lgroup and PPC 1?xmol/L?5?mol/L?10?mol/Lgroup,cell viability is determined by MTT method.The second part:according to the result of determined by MTT,selecting the optimal concentration of GSH and PPC drug group,set up blank control,collect training 1,3,5,7 days of supernatant fluid,detection of the variation in content of albumin,cereal third transaminase,aspertate aminotransferase.Results:1.Improved collagen perfusion method to isolate the total number of hepatocytes by more than 109;2.The number of people who have been able to measure fresh separation from the blue dye method is more than 90 percent.3.The results showed that the GSH5mM group and the PPC5 ?mol/L group were the most effective.4.Test 1-7 days of blank group that albumin content in the supernatant fluid was no significant difference(P>0.05),GSH and PPC groups that albumin content in the supernatant fluid increases obviously,P<0.05,In the 7th day,the albumin content was greater than the blank control group(P<0.05);5.In group 1,3,5 days blank ALT and AST in the extracellular fluid content significantly greater than drug group,the results have significance(P<0.05),5 days before the ALT and AST in extracellular fluid leakage to reduce gradually,reached the lowest in 5 days,after the content.The results were no different in the seventh day(P>0.05).Conclusion:1.In the 1L medium of culture,the addition of the prototype glutathione 5mM and polyene phosphatidylcholine 5?M has a good protective effect on the active liver cell activity of the adult in vitro culture.2.The reduction of the good albumin secretion of the adult proto-hepatocytes,which are isolated and cultured by the prototypal glutathione and polyenophosphates.3.The prototype glutathione and polyene phosphatidylcholine can maintain the cell membrane integrity of adult liver cells.