Effects Of Glutamine On The Growth And Metabolism Of Carp Intestinal Epithelial Cells In Primary Culture

The carp intestinal epithelial cells in primary culture were selected to evaluate the trophic actions of glutamine. Firstly, the isolation and primary culture method of carp intestinal epithelial cells was developed. The carp intestinal epithelial cells in primary show a normal crypt cell characteristic by examination of morphologic characterization. And the carp intestinal epithelial cells growth characteriztion was investigated. The primary culture carp intestinal epithelial cells attached within 48-72h proliferated obviously at 120-168h.These results shows that this culture conditions is suitable for the study of effects of glutamine on the carp intestinal cell.Secondly, the cell MMT absorption at 540 nm wavelength, alkaline phosphatase activities, creatine phosphokinase activities, alanine aminotransferase activities, glutamine utilization, ammonia production, glucose utilization, lactate production, protein utilization, protein retention were investigation when the carp intestinal epithelial cells was growing medium of containing 0, 300, 700, 800,1000, 1200 and 1400 mg/L glutamine respectively. The results showed that MTT absorption at 540 nm wavelength (MTT OD₅₄₀) , alkaline phosphatase activities, glutamine utilization, protein utilization, protein retention, creatine phosphokinase activities and alanine aminotransferase activities were very significantly affected by glutamine concentration (P < 0.01). The effects of glutamine concentration on the ammonia production, glucose utilization, lactate production were not significant (P>0.05).The MTT OD₅₄₀, alkaline phosphatase activities, glutamine utilization, protein utilization and protein retention were increased with the glutamine level increasing. But the creatine phosphokinase activities and alanine aminotransferase activities were decreased with the glutamine level increasing. The results of correlation analysis showed that there were significant correlation between MTT OD₅₄₀, alkalinephosphatase activities, protein utilization, protein retention, creatine phosphokinase activities, alanine aminotransferase activities and glutamine utilization (rl-+0.9463, P < 0.01; r2- +0.9823,P < 0.01; r3- +0.9201,P < 0.01; r4- +0.8978,P < 0.05; r5- +0.9371, P < 0.01, rl—0.8983, P < 0.05; r2—0.8502, P < 0.05; r3⁰.9930, P < 0.01). The regression results showed that there were very significant quadratic relationship between MTT ODho glutamine level (R2-0.9762, P<0.01) . In summary, glutamine could enhance proliferation and differentiation of the carp intestinal cells in primary culture, improve protein retention and decrease the appearance of epithelial death. So glutamine has a stronger effect on the carp epithelial growth in primary culture and is a very important specific nutrient to the intestinal mucosa of fish.