The Effect Of Ulinastatin Injection On Treg,TH17 Lymphocyte Subsets In The Sepsis Rats

There are sorts of severe diseases contributing to the sepsis. The sepsis can exasperate and become sepsis shock that lead to the multiple organ failure(MOF). On the same time, the high medical cost is a huge burden for the family and the society. Although it has been affirm to be helpfulness for the patient to be resuscitated and treated with the antibiotics in the early stage, the mortality of the sepsis constantly maintains at a high level. Many researchers try their best to explore the reason. Finally, they find that the infection is one of the major reasons for the deterioration of sepsis. In the severe cases, the optimal antibiotics which therapists utilize cannot effectively kill the organism. It is essential to explore the working mechanism and the dynamic change of the immune system in the septic patients. The immune system contains B cell,T cell and so on. Th1,Th2 are the traditional T cell subset,their balance relationship directly affect the immune function. Now,people have discovery two sorts of new T cell subset, Treg and Th17.They all derive from primitive T cell and can be compensation for the theory of Th1,Th2.Treg can affect the polarization of Th1/Th2/Th17 and itsspecific transcription factors is Foxp3;Th17 could secret specific IL-17 and Its specific transcription factors is ROR?t.Acidoglycoprptein(AGP) which is produced within the liver,called ulinastatin,is a immunologic regulator and enzyme inhibitor which is egained from male human tale.The substance is able to ameliorate severe inflammatory response, such as decreasing the expression of TNF-α and IL-6 and promoting the number of B cell. It can also make the paralysed immune function stable. But there is few research working out how it effect Treg and Th17 cell.The study utilized CLP to copy the sepsis model. We hoped to explore the reason why the Th17 and Treg cell changed dynamical and the effect that the Th17 and Treg cell affected the immune system. Besides,we wanted to research ulinastatin’s immunology regulatory function.In this subject,the animal models were reproduced by CLP. There were four groups(normal group, sham group, sodium chloride group and ulinastatin group)involved in it. Each group contained eight Male Wistar( 250 — 300g)rats. After finishing the surgeon, 30mL/kg of normal saline was injected in the subcutaneous tissue to compensate the loss of body fluid and blood volume. 5ml blood and the spleen in each rat were harvested from the vessels after surgery. We studied the fluctuated expression of the proportion of Treg、Th17 in all the lymphocyte and the Treg/Th17 in the spleen with the the flow cytometry(FCM); The changeof specific transcription factors ROR? t and Foxp3 mRNA in spleen tissue would be researched with the RT-PCR; We utilized ELISA to study the change of the IL-17 in the plasma; immunohistochemical experiment were used to detect the expression of CD4+T cell, CD8+T cell expression.The flow cytometry result show : Compared with normal, sham group,the proportion of Treg,Th17 in spleen tissue in NS group(P <0.01)were increased in the late stage of sepsis. Compared with NS group,the proportion of Treg,Th17 in spleen tissue in UTI group(P <0.01)were decreased in the late stage of sepsis. Compared with Normal, sham group,there was indifference of the Treg/Th17(P >0.05) in NS group.Compared with Normal, sham group, Treg/Th17( P <0.05) was decreased in UTI group;RT-PCR result show:the levels of Foxp3,ROR?t mRNA in spleen tissue(P <0.01)in NS group were higher than Normal, Sham, UTI group.There was no difference in the Foxp3,ROR?t mRNA expression between the UTI group and the Normal,sham group(P >0.05);ELISA results show: the levels of IL-17 in plasma in NS group(P<0.05) were higher than Normal, Sham, UTI group. There was no statistics significance in the IL-17 expression between the UTI group and the Normal,sham group;Immunohistochemical result show: Under the 200 multiple microscope,the brown lump which present the CD4+cell is lesser than theNormal,Sham, UTI group. There was no significant difference in the CD8+cell expression between Normal, Sham, NS and UTI group respective.In summary, the sepsis disrupted the immune balance.There were not only promoted inflammatory response but also immunosuppression.The elevation of Th17 expression was characteristic of the promoted inflammatory response. The high expression of Treg contributed to the immune paralysis and the low expression of the CD4+ T cell. ulinastatin could alleviate the inflammatory response and avoid the immune paralysis, promote the number of CD4+T cell, ameliorate the immunosuppression.