Study On The Therapeutic Effect And Mechanism Of Huperzine A On Sepsis

Huperzine A(HupA),as a highly effective,reversible and selective acetylcholinesterase inhibitor(ACh EI),is mainly used to treat neurological diseases such as dementia,to enhance learning and memory,and improve spatial memory disorders.There are indications in existing studies that it has a certain anti-inflammatory effect.Sepsis,as a systemic comprehensive disease involving multiple organs and systems,has a complex pathogenesis.Exploring effective treatment drugs has become a hot and difficult point in the current research on sepsis.Therefore,this thesis explores the therapeutic effect and the underlying mechanism of HupA on sepsis by testing an animal model of sepsis.This article first validates the anti-inflammatory effects of HupA in lipopolysaccharides(LPS)-induced inflammatory mice.We found that HupA can significantly improve the survival rate of mice.At a dose of 0.1 mg/kg HupA injection,the survival rate of mice increased to 40%.The serum inflammatory factor expression of mice was measured by enzyme-linked immunosorbent assay(ELISA).HupA significantly reduced the expression of inflammatory factors interleukin-6(IL-6),tumor necrosis factor(TNF-α)and interleukin-1 beta(IL-1β)in inflammatory mice;at the m RNA level,HupA can reduce the inflammatory factors IL-6,IL-1βexpression in the kidneys and lungs of LPS-induced inflammation mice,but there was no significant difference in the expression of IL-6 and IL-1βin liver tissues.Based on the above data,we further verified whether HupA has the same therapeutic effect on cecal ligation and puncture(CLP)sepsis mice.By ligation and puncture of the cecum of the mouse,the lethality of the mouse reached 90%within three days,and the survival rate of sepsis mice was effectively increased to 40%under the treatment of HupA at 0.1 mg/kg,HupA at 1 mg/kg and 0.01 mg/kg only increased the survival rate of sepsis mice to 20%.Because this experiment performed cecal ligation and puncture in mice and caused the contents to flow out,we chose TIENAM to control the growth of microorganisms which might be resulted from this outflow.Among them,the combined survival rate of sepsis mice was further increased by 50%.Using enzyme-linked immunosorbent assay,real-time fluorescence quantitative RT-PCR,hematoxylin-eosin(HE)staining and other methods to examine the physiological and pathological conditions of sepsis mice,it can be seen that the inflammatory factor IL-6 and high mobility group box-1(HMGB-1)in serum of CLP sepsis mice under the combined action of HupA and TIENAM were significantly down-regulated,and the expressions of serum enzymes such as amylase(AMY),alanine aminotransferase(ALT),and alkaline phosphatase(ALP)were significantly down-regulated.HE staining of the lung and liver showed that the combination of HupA and TIENAM could alleviate the infiltration of inflammatory cells and organ lesions in the lung and liver tissues of sepsis mice.In order to study the role of HupA in the regulation of immune disorders during the pathogenesis of sepsis,we used flow cytometry to study changes of the T lymphocytes and B lymphocytes in the blood of sepsis mice.The data showed the increases of CD3~+T,CD4~+T,and CD8~+T cells,but a decrease of B cells in sepsis mice.HupA treatment led to the recovery of T lymphocyte expression to the sham operation level,but B cell expression was not significantly affected.Therefore,the immune regulation of HupA in sepsis mice is likely to take place in T lymphocytes,but the specific mechanism needs to be further studied.In this study,the protective effects of HupA on inflammation and sepsis were studied in both inflammatory and sepsis mice.The further studies regarding the HupA mechanism of its action and the possibility of HupA used as a drug for sepsis need to be further explored,so that“new use of old drugs”will become the new trend of future drug research.