Effects Of Composite Additives On The Anti-Transport-Stress Ability Of Rats And Optimal Dose Prescriptions Selection

The purpose of this study was to investigate the effects of anti-stress composite additives on rats undersimulated transport stress, and hope to provide a reference for the development of new anti-stress compositeadditives.In the present study, take the clean grade wistar male rats as the study object which was12weeks of ageand weight was （250±20）g. Therefore,110rats were randomly divided into three groups by L49（3）. Thetreatment group was intragastric administration mixture of VC, VE, KHCO₃and GABA（suspending by CMC）.Other two control groups was intragastric administration0.5%CMC solution. A week later, the rats oftreatment group and stress-control group were shaked in a shaking table with rotational speed180r/min tosimulated transport stress. Eleven stress indicators （CK, LDH, GLU, ACTH, GC, IL-2, IL-4, IL-6, TNF-α andHSP70）levels in the serum of rats were analyzed to evaluate the efficacy of anti-stress composite additives.The results showed that the indicators have varying degrees of elevated after2hours transport stress, and theadditives could suppress the elevated levels of CK, ACTH, GC, IL-2, IL-4, IL-6and HSP70. The above resultsproved that the self-designed anti-stress composite additive has a good effects on resistant transportation stress.Then further data analysis by principal component analysis （PCA） and consolidated balance method. Theresults were showed as following:（1） Effects of anti-stress composite additives on GLU, enzymes and hormones of rats under simulated transportstress.The GLU generally tended to increase after2h of transport stress, group5and group7GLU level withthe negative control group showed no significant change （P>0.05）, but the group7showed significantlydifference compared with positive control group （P<0.05）, the other groups compared with the positive controlgroup was not statistically significant difference （P>0.05）. The CK concentration of rats of positive controlwas extremely increased significantly compared with group1,3,4,6,7and negative control group （P<0.01）,and increased significantly compared with group9（P<0.05）. The LDH concentration test results showed thatthe negative control group and group1was extremely significant difference compared with the positivecontrol group（P<0.01）, positive control group showed a significant differences compared with the group4（P<0.05）. The negative control group increased extremely significant of the ACTH concentration comparedwith the group4（P<0.01）, the positive control group increased significantly compared with the group3,5,6,7and group9（P<0.05）. The GC concentration test results showed that the group4and negative control groupshowed significant difference compared with the positive control group （P<0.05）, the other groups comparedwith the positive control group had no significant difference （P>0.05）.（2） Effects of anti-stress composite additives on related cytokines of rats under simulated transport stress.After transport stress, the IL-2concentration of negative control and group1,2,4,5,6was significantlylower than the positive control group （P<0.05）. Concentration of IL-4in the positive control group wasextremely significantly higher than negative control group and the group4（P<0.01）, and it was significantlyhigher than group3and group5（P<0.05）, and other groups have no obvious changes （P>0.05）. The IL-6concentration of the positive control group was significantly higher than group2（P<0.05）, and there was nosignificant change between the other groups and the positive control group（P>0.05）. TNF-α concentration test results showed that the positive control group was significantly higher compared with negative control groupand group3（P<0.05）, and there is no significant difference with the other groups （P>0.05）.（3） Effects of anti-stress composite additives on HSP70expression of lymphocyte of rats under simulatedtransport stress.The HSP70mRNA level of q-PCR test results showed that the positive control group was not significantdifference compared with group7（P>0.05）, and compared with the other groups showed increased extremelysignificantly （P<0.01）. After transport stress, the HSP70expression changes showed as follows, the positivecontrol group increased extremely significantly compared with negative control group （P<0.01）, and increasedextremely significantly compared with group2and group4（P<0.01）, and increased significantly comparedwith group1,5and8（P<0.05）.（4） Selection of optimal formula for anti-transport-stress additives.The raw data of the11indicators were analysis by principal component analysis （PCA） and consolidatedbalance method. The results showed that the best intragastric administration dose were VC10mg/kg, VE5mg/kg, GABA3mg/kg, KHCO₃15mg/kg.The above results proved that the self-designed anti-stress compound additive has a good effects onanti-transport-stress. The best formula of anti-transport-stress additives were VC200mg/kg, VE100mg/kg,GABA60mg/kg, KHCO₃300mg/kg.