Effects Of Glutamine On Apoptosis In Rat Intestinal Mucosal Cell In Cold Restrain Stress

Objectives: To observe the change of apoptosis in intestinal mucosal cellsinduced by CRS (cold restrain stress ), and discuss the effect of glutamine and its pathogenesis on the apoptosis induced by the stress.Materials and Methods: The SD rat were divided into three groups atrandom, i.e, normal group, stress group and glutamine group. The stress model was made in terms of the model made by Brodie in 1969. The sample were 5cm long collected from the end of ileum. Biopsy specimens were processed for routine histological examination and immunohistochemical evaluation of PCNA and Bcl-2 expression. For the detection of apoptosis in situ, terminal deoxynucleotidyl transferase-mediated nick end labeling staining (TUNEL) was used. Spectrophotometric method was used to detect the blood concentration of glutamine.Results:1. The analysis of experimental data among the three groups, i.e, N group (n=10) , S group (n=25) and G group(n=25).1.1 Comparsion of the apoptotic rate (AR) of intestinal mucosal cells among the three groups.The experimental results indicate that the apoptotic cell mostly localized on the top of the vili of small intestine ,the crypt and inherent layer also have a few ones.It is very apparent that the AR of all unit of the stress is higher than that of controls (P<0.05). Among them the 4h unit is the highest one.but comparing with the AR of the upper corresponding unit of the two group,the AR of the G group is lower than that of S group(p<0.05). However, the AR of the 12h and 24h unit of the G group is all close to that of N group(P>0.05).1.2 Comparison of the blood concentration of glutamine (CG) among the three groups.the data show that CG of all unit of the stress and glutamine group is lower than that of controls, Among them the 4h unit is the lowest one. but comparing with the CG of the upper corresponding unit of the two group,the CG of the G group is higher than that of S group(p<0.05). However, the CG of the 12h and 24h unit of the G group is all close to that of N group(P>0.05).1.3 The comparison of the expression of the PCNA among the threegroupsThe immunohistochemical method was used to detect the expression of the PCNA in intestinal mucosal cell. It took on over the moderate positive result in the most of the three groups except for the 2h and 4h among S groups.The result of quantitative analysis show it is between 12% ~ 15%,there was no obvious difference in statistics(P>0.05).1.4 The comparison of the expression of the BcI-2 among the three groupsThe same method was used as the detection of PCNA, Among the three groups,the S2h,S4h,S8h and G2h,G4h group have no expression of Bcl-2 ,While the rest of other groups all have obvious expression.,N group also have little expression of Bcl-2. 2. The analysis of correlation between Gin and each experimental item2.1 The relation between the blood concentration of glutamine (BCG)and the apoptic rate.From the result above we can see there were negative correlations between them (r=-0.354,P<0.05).With the decreasing and increasing of the concentration of the glutamine ,the AR increase and decrease .Moreover, the group with the lowest BCG has the highest AR . It is very easy for us to conclude that the deficiency of glutamine can increase the apoptosis of intestinal mucosal cell. While supplement of glutamine can depress it.2.2 the relation between the blood concentration of glutamine (BCG)andthe expression of PCNAIn stress conditions, the expression of PCNA will not reduce until the BCG decreased very intensively, therefore we can conclude that during the state of stress, the Gin affect little on the expression of PCNA(r=O.127,P>0.05).2.3 the relation between the blood concentration of glutamine (BCG)andthe expression of Bcl-2.Bcl-2 almost have no expression when the BCG decreased . but supplement of Gin can increased its expression intensively. The fact tell usthat Bcl-2 is one of the pathogenesis through which Gin depress the apoptosisof intestinal mucosal cell.Conclusions: Besides the common gastric mucosal pathological changes,the stress can lead to the increasing apoptosis of intestinal mucosal cell, so itwill damage the intestinal barrier function. But Glutamine can depress theincreasing apoptosis of it .protect the intestinal barrier.and affect little on itsproliferation...