Effect Of The Different Dose Of Thymosin Alpha 1 On The Inflammatory Response In The Mice Of Sepsis Induced By SEB And LPS

Objective: To evaluate the effect of the different dose of thymosin alpha 1 on prognosis of sepsis,This study compared the effect of the high-dose thymosin alpha 1 and the low-dose thymosin alpha 1 on the concentration of serum INF-?,the expression of NF-?B p65 in the spleen and the change of the pathological tissue in the mice of sepsis induced by SEB and LPS.Methods: 1 The establishment of animal model 1.1 The preliminary experiment were explored by using different doses of SEB and LPS to establish experimental mice of sepsis.1.1.1 Experimental groups: The 80 mice were randomly divided into 10 groups,each group had 8 mice.1)Induced by the dose of 500?g per kilogram SEB and 1500?g per kilogram LPS2)Induced by the dose of 500?g per kilogram SEB and 1000?g per kilogram LPS3)Induced by the dose of 500?g per kilogram SEB and 500?g per kilogram LPS4)Induced by the dose of 400?g per kilogram SEB and 1500?g per kilogram LPS5)Induced by the dose of 400?g per kilogram SEB and 1000?g per kilogram LPS6)Induced by the dose of 400?g per kilogram SEB and 500?g per kilogram LPS7)Induced by the dose of 300?g per kilogram SEB and 1500?g per kilogram LPS8)Induced by the dose of 300?g per kilogram SEB and 1000?g per kilogram LPS9)Induced by the dose of 300?g per kilogram SEB and 500?g per kilogram LPS10)Induced by the same dose of PBS 1.1.2 The experimental groups received different doses of SEB which was dissolved by PBS,and the control group received 0.2ml PBS By intraperitoneal injection.1.1.3 The experimental groups received different doses of LPS which was dissolved by PBS after 2 hours,and the control group received 0.2ml PBS By intraperitoneal injection.1.1.4 The symptoms and survival of 72 h in mice were observed.1.2 There was no symptom in the control group within 72 hours.Within 4 hours,abnormal symptoms appeared in the group of combined application of SEB and LPS.However,there was a series of abnormalities such as piloerection,shortness of breath,listlessness,not eating and diarrhea in the other experimental groups of mice.There was only one mouse surviving in the group induced by the dose of 500?g per kilogram SEB and 1500?g per kilogram LPS.There were only two mice surviving in the groups induced by the dose of 500?g per kilogram SEB and 1000?g per kilogram LPS and induced by the dose of 400?g per kilogram SEB and 1500?g per kilogram LPS.There were three mice surviving in the group induced by the dose of 500?g per kilogram SEB and 500?g per kilogram LPS.There were five mice surviving in the group induced by the dose of 400?g per kilogram SEB and 1000?g per kilogram LPS.There were six mice surviving in the group induced by the dose of 400?g per kilogram SEB and 500?g per kilogram LPS.And all mice of the left three groups survived.The symptoms of the groups induced by the dose of 300?g per kilogram SEB and 500?g per kilogram LPS were relatively light,and the symptoms of the group relieved,and the mice began to eat after six hours.The symptoms of the group induced by the dose of 300?g per kilogram SEB and 1000?g per kilogram LPS were worse after 3 hours to 8 hours,and the symptoms improved after 24 hours,and the mice recovered after 48 hours.While the symptoms of the group induced by the dose of 300?g per kilogram SEB and 1500?g per kilogram LPS improved after 48 hours,and the mice recovered after 64 hours.According to the experimental results above,the model induced by the dose of 300?g per kilogram SEB and 1000?g per kilogram LPS was selected,and the process of the model was repeated over and over,finally,the model of sepsis was established.2 Age BALB/c mice,male,about 19g-21 g weight,were received the dose of 300?g per kilogram SEB and 1000?g per kilogram LPS by intraperitoneal injection to establish the model of sepsis.The mice were divided into the control group,the high-dose group and the low-dose group at random.The low-dose group was given 100?g per kilogram dose of thymosin alpha 1 by subcutaneous injection.And the high-dose group was given 2000?g per kilogram dose of thymosin alpha 1 by subcutaneous injection.The control group was given the same dose of PBS.We compared the expression level of serum cytokines in each group.The expression of serum INF-? was measured by ELASA,and the expression of spleen NF-?B p65 was measured by Western blot.After 6 hours,the lung tissue of every group was stained with HE,and the pathological changes of lung tissue were compared.Results: Within 4 hours after intervention,all the groups of mice appeared a series of abnormalities such as piloerection,shortness of breath,listlessness,loss of appetite,diarrhea,loss of interest in the surrounding environment,self-cleaning behavior disappearing and the secretion of eyelid increasing.While the symptoms of the high-dose group and the low-dose group were relative light comparing to the control group.After 5~6 hours,the high-dose group and the low-dose group showed signs of mental improvement and self-cleaning behaviors,while the control group did not restore.There was a rise in the serum levels of interferon gamma and spleen of NF-?B p65 at 2 hours comparing the high-dose group and the low-dose group to the control group,the difference was statistically significant(P< 0.05),there was no significant difference between the low-dose group and the high-dose group(P>0.05).There was no statistical significance at 4 hours comparing the low-dose group and the high-dose group to the control group in the serum levels of INF-? and in the level of pleen NF-?B p65(P>0.05).There was no statistical significance at 6 hours comparing the low-dose group and the high-dose group to the control group in the serum levels of INF-? and in the level of pleen NF-?B p65(P>0.05).At 6 hours,the damage of alveolar wall and the hemorrhage pulmonary interstitial was visible in the control group.The low-dose group showed pulmonary interstitial vascular dilatation and congestion but no hemorrhage.The injury of the lung in the high-dose group was mild comparing with the control group.But there was no significant difference between the high-dose group and low-dose group.Conclusion:1 In the early inflammatory response,thymosin alpha 1 up-regulated the expression of serum INF-? and NF-?B p65 of the spleen,and the immune response of the mice was enhanced.2 No significant effect was observed between the high-dose thymosin alpha 1 and the low-dose thymosin alpha 1 on the early inflammatory response of sepsis in mice.