| In this study,an equipment of simulating transportation was used to set up a model of transport stress in rats.Then,the effects of oral administrated conditionally essential amino acid glutamine on amino acid profile,related biochemical indexs in serum and tissue and HSP mRNA expression in muscle were investigated using this model.On the base of these, the protective mechanism of glutamine on transport stress was discussed.The results of this study made further understand of the biological effect of glutamine and provided new ideas of defend transport stress for animals.1.Determination of Free Amino-acids in Serum and Tissues of Rats by Reverser-phase High Performance Liquid ChromatographyA pre-column deviation procedure of O-Phthaldialdelhyde(OPA) was used for the determination of amino-acids.Amino-acid derivatives were separated on Waters Spherisorb ODS(4.6mm×250 mm,5μm) column.Gradient elution was adopted with fluorescence detection.Fifteen amino-acids were separated and detected within 40min.Linear relationship between the peak area and concentration in the range of 5.20 to 333nmol·L-1 of the amino-acids,with correlation coefficients of 0.9993 to 1 was obtained.Amino-acids with the intra-day and inter-day RSD are 2.01%-5.53%and 3.65%-5.88%,respectively.The lowest detect area is 1.2nmol·L-1.The amino-acids recovery of serum,hypothalamus liver, muscle,heart and kidney are in the range of 107.10%-122.28%,90.90%-110.95%, 103.43%-119.95%,90.33%-111.30%,93.82%-126.51%,90.39%-144.15%,respectively.2.Effect of Glutamine on Amino acid Profile of Serum and Tissues in Rats under Simulating Transport StressIn this study we studied the anti-stress effects of glutamine on rats suffered from simulating transporting stress.70 male SD rats were randomly divided into negative control ((-)10rats),low frequency transport stress(4Hz) group(30rats) and high frequency transport stress(6Hz) group(30rats).30 rats of each frequency stress group also included positive control group(+),Phenobarbital group(P) and Gln group(Q),10 rats of each group. Each rat in(-) and(+) group was oral administered with physiological saline(1 mL/day). The rats of P and Q groups were oral administered with Phenobarbital(15mg·Kg-1/day) and Gln(500mg·Kg-1/day) for 10 days respectively.After 10 days,rats in each group were vibrated with simulated low(L)and high frequency(H) transport stress for 2 hours except for(-)group,then all the rats was killed by exsanguinations.Serum,hypothalamus,liver and muscle samples were collected and tested for the content of dissociative amino acid in blood and these tissues by RP-HPLC.The results showed that compared with(-)group, significantly differences of glycogenic amino acids,aromatic amino acids and branched-chain amino acid in serum,hypothalamus and muscle could not be found in rats treated with Gln(P＞0.05),but the content of glycogenic amino acids,aromatic amino acids and branched-chain amino acid was obviously higher than(+) group(P＜0.05 or P＜0.01). And the narcotic of Phenobarbital did not show any effects on resisting transport stress.The results of this study demonstrated that Gln can protect rats from transport stress.3.Effect of Glutamine on Related Biochemical Indexes and HSP72mRNA Expression in Rats under Simulating Transport StressThe present study detected the anti-stress effects of glutamine on rats suffered from simulating transporting stress.70 male SD rats were randomly divided into negative control ((-)10rats),low frequency transport stress(4Hz) group(30rats) and high frequency transport stress(6Hz) group(30rats).30 rats of each frequency stress group also included positive control group(+),Phenobarbital group(P) and Gln group(Q),10 rats of each group. Each rat in(-) and(+) group was oral administered with physiological saline(1 mL/day). The rats of P and Q groups were oral administered with Phenobarbital(15mg·Kg-1/day) and Gln(500mg·Kg-1/day) for I0 days respectively.After 10 days,rats in each group were vibrated with simulated low(L) and high frequency(H) transport stress for 2 hours except for(-) group,then all the rats was killed by exsanguinations,and blood,liver and muscle samples were collected for testing Creatine Kinase(CK),Lactate Dehydrogenase(LDH), blood glucose,muscle glycogen and hepatic glycogen.The results showed that no significant difference of CK,LDH,blood glucose,muscle glycogen and hepatic glycogen in serum,liver and muscle was observed in rats treated with Gln compared with(-)group (P＞0.05),HSP72 mRNA expression in skeletal muscle of L(+),H(+),LQ and HQ group increased significantly(P＜0.05 or P＜0.01).And the narcotic of Phenobarbital did not exhibit any transport stress resistance effects.The results of this study demonstrated that Gln had anti-stress protective effects on transporting rats.
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