The Effectof Asperosaponin Ⅵ On Protecting Mice Fromsepsis And Its Mechanisms

Sepsis,associated with multiple organ dysfunction,is a systemic inflammatory response syndrome caused by infection.At present,supportive therapy and antibiotic application is still the mainstream treatment of sepsis.However,due to its complex pathogenesis,the morbidity and mortality of sepsis have been high,and it has become the main cause of death of patients in intensive care,and has also been the focus of clinical medicine.Once sepsis is not timely intervened,the early state of high inflammation will evolve into the subsequent state of low inflammation,and it is easy to cause severe septic shock or even multiple organ dysfunction.Asperosaponin Ⅵ(AⅥ)is a triterpenoid saponin compound,which has been proved to have obvious anti-inflammatory and immune effects on inflammatory diseases such as neuritis,myocarditis and osteoporosis.However,the role of AⅥ in sepsis and its potential mechanism remain unclear.In this study,cecal ligation puncture(CLP)-induced sepsis and LPS-induced sepsis models were used at the same time to investigate the effect of AⅥ on sepsis mice and its possible mechanism.Methods:1.In vitro experiment,bone marrow mononuclear macrophages(BMMs)and THP-1 derived macrophages were stimulated by LPS with pretreatment of AVI and UTI for 24h.The experimental groups were divided into Control group(Blank),LPS model group(LPS),LPS+Asperosaponin Ⅵ group(AⅥ)and LPS+Ulinastatin positive drug group(UTI).The protective effect of AⅥ on LPS-induced inflammation was detected by real-time fluorescence quantitative PCR and Western blot.2.In vivo experiment,the effect of AⅥ on sepsis and its mechanism was investigated by CLP-and LPS-induced sepsis model.CLP-induced sepsis model animals were divided into Control group(Blank),CLP model group(CLP),CLP+Asperosaponin Ⅵ low-dose and high-dose group(A10:10mg/kg/d,A20:20mg/kg/d)and CLP+Ulinastatin positive drug group(UTI:5×104U/kg).LPS-induced sepsis model animals were divided into Control group(Blank),LPS model group(LPS),LPS+Asperosaponin Ⅵ low-dose and high-dose group(A10:10mg/kg/d,A20:20mg/kg/d)and LPS+ Ulinastatin group(UTI:5×104U/kg).Intragastric administration for 14 days,once a day,followed by surgery.The changes of anal temperature and behavior were observed after operation.The bacterial content of peritoneal lavage fluid and the content of Aspartate aminotransferase(AST),Alanine aminotransferase(ALT),Lactate dehydrogenase(LDH)and Lactate dehydrogenase(LD)in plasma were determined.Plasma inflammatory factors were detected by ELISA.Evaluate the dry/wet weight ratio of lung tissues.At the same time,the liver,spleen and lung of mice were stained with H&E.Western blot,real-time fluorescence quantitative PCR and immunohistochemistry were used to detect signaling pathways in spleen and lung tissues.Results:1.In vitro,compared with LPS group,pretreatment of AⅥ and UTI could significantly inhibit the expression of inflammatory factors.It can inhibit the occurrence and development of inflammation by regulating Hippo signaling pathway and Rho family.2.In vivo experiment,both UTI and AⅥ could inhibit the expression of inflammatory factors in CLP-and LPS-induced sepsis mice,and reduce the bacterial content in abdominal lavage fluid.In addition,pretreatment with AⅥ and UTI alleviated liver,spleen and lung injury caused by sepsis,and reduced inflammatory response.Moreover,these protective effects are mediated by inhibition of Hippo signaling pathway and the Rho family.Conclusions:1.Asperosaponin Ⅵ has a protective effect on sepsis mice,can inhibit the inflammatory response of sepsis mice,and reduce the bacterial content in the peritoneal lavage fluid of mice.2.The preventive treatment of Asperosaponin Ⅵ can alleviate organ damage caused by sepsis and reduce inflammatory reaction.3.In sepsis model,Asperosaponin Ⅵ can significantly protect mice from damage and inhibit Hippo signaling pathway and Rho family.