| Septic shock is the widespread and complex problem concerned in modern Critical Care Medicine.It becomes the prominent issue to seriously impact on prognosis of patients and further improvement of successful treatment rate.One group of abroad clinical epidemiological research papers indicated that septic shock occurred at a rate of 4%among 2527 cases of patients with systemic inflammatory response,and 46%of septic shock patients were died.A study in China showed that 40%patients whose body surface areas are burned more than 30 percent had concurrently septic shock.Study on critically ill patients also showed that incidence of septic shock was 15.7%,61.1%of septic shock further developed to multiple organ dysfunction syndrome(MODS),fatality rate was 30.6%.Based on above studies,it has very important theoretical value and clinical significance to reveal septic shock pathogenesis and explore clinical prevention and treatment methods.When simplex western medicine treatment hardly makes progress in effect of septic shock,it appears urgently to look for Western & Chinese Traditional Medicine method to treat septic shock.Many medical scholars hope to find a breakthrough for septic shock with Chinese traditional medicine treatment.According to clinical manifestations, the septic shock is the scope of syncope.As most clinical manifestations are also collapse,it's also named as syncope and collapse syndrome.The development of traditional Chinese medicine in thousands of years was a research on septic shock and accumulated much experience,but the treatment is lack of evidence because of the unclear mechanism of septic shock.Much progress has been made in recent years through studying septic shock,however brain injury is rarely involved.This study takes the models of endotoxin-shock rats as research object to simulate septic shock.The study aims are to explore anomalous signal transduction in brain injury caused by septic shock and the effect of Sini Decoction in the process;define the molecular and cellular mechanisms of reviving yang for resuscitation method used to prevent and treat brain injury in septic shock and establish the scientific foundation for modernizing traditional Chinese medicine.Objective1.Clear level of inflammatory mediators,the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the changes of function metabolism and structural of brain vascular smooth muscle cells,glial cells,nerve cells stimulated by the extracellular inflammatory mediator of brain injury in septic shock.Then sum up regular relationship among them.2.Clear if the role of increasing of IL-1α,TNF-αcan upregulate the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the relationship between the abnormal quantity of gas messenger-NO in brain vascular smooth muscle cells,glial cells and nerve cells and different brain regions vascular response and neuronal damage and repair.3.Clear how Sini decoction act on above indicators in order to descript the method of reviving yang for resuscitation how improve molecular mechanisms of brain microcirculatory disturbances and neuronal damage.Develop Chinese medicine A&E theory.Methods1.This study takes the models of endotoxin-shock rats simulating septic shock for research object.First using technology of gene chip to screen prereceptor signal substances, inflammatory cytokines,which have an obvious role in septic shock.2.To explore the role of Sini decoction on prereceptor signal substances induced by septic shock.Such as levels of tissue inflammatory mediators,IL-1α,TNF-α.The specific methods uses ELISA to measure IL-1α,TNF-αcontent in septic rat brain tissue.3.To explore how Sini decoction act on receptor signal substances induced by increasing IL-1α,TNF-αcontent.Such as the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,ICAM-1(CD54),NCAM(CD56),VCAM-1(CD106). The specific methods uses SABC to determine distribution and quantity of ICAM-1(CD54),NCAM(CD56),VCAM-1(CD106) in rat brain tissue.4.To explore how Sini decoction act on the changes of signal substances after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation. Such as NO production and SOD,MDA content in rat brain.The specific methods is to use biochemical methods to determine SOD,MDA content and SABC to determine distribution and quantity of iNOS rat brain tissue.5.At last explore how Sini decoction improve tissue structure after septic brain injury. Using TUNEL to determine apoptosis in rat brain,ultrastructural changes of the cells with electron microscopy,structural changes of brain tissue with microscopy.Results1.Mean arterial blood pressure of sham-operated rats started to decline slightly at 1 hour. After i.v.injection of lipopolysaccharide,significant difference between model group and sham operated group at 1 hour(p<0.01),blood pressure fell to 40.75%of the initial values at 6 hour. Blood pressure of dexamethasone group fell to 45.32%of the initial values at 6 hour and had higher blood pressure compared with model group(p<0.05).Blood pressure of Sini decoction group fell to 67.68%of the initial values at 6 hour had higher blood pressure compared with model group(p<0.01).2.Screening Ccl2,4,7,20,Cxcll,2,9,10,11,Ccr2,3,9;TNF-α,IL-1α,IL-1βwith very significant high expression in endotoxic rat brain.And some inflammatory cytokines such asIL-13,IL- 15,I15ra which were very few reports had differentially expressed. IL-13 has no changes at 1 hour,up-regulation at2,3,6 hour;IL-15 has down-regulation at 1,2 hour and I15ra has down-regulation at 1,2,3,6 hour. 3.Concentration changes of TNF-α,IL-1αin endotoxic rat brain were same as mRNA content changes detected by gene chip technology from 1 hour to 6hour.①Concentration of TNF-αin model group were higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.Concentration of TNF-αof dexamethasone group were lower than model group from 1 hour to 6 hour(p<0.01).Concentration of TNF-αof Sini decoction group were lower than model group at 3,6 hour(p<0.01).②Concentration of IL-1αin model group were lower than sham operated group at 1 hour(p<0.01),no significant difference at 2 hour(P>0.05),higher at 3(p<0.05),6(p<0.01)hour and reached peak at 6 hour.Concentration of IL-1αof dexamethasone group were lower than model group from 1 hour to 6hour(p<0.01). Concentration of IL-1αof Sini decoction group were lower than model group at 2,3,6 hour(p<0.01).4.No expression of CD54 and weak expression of CD56,CD106 in normal rat brain.The expression of CD54,CD56,CD106 in model group was higher than sham operated group and control group on four time periods(p<0.01).CD54,CD106 reached peak at 1 hour and CD56 reached peak at 6hour.The CD54 expressions of dexamethasone group were lower than model group at 1,2 hour(p<0.01).The CD56 expressions of dexamethasone group were higher than model group at 1 hour(p<0.01).The CD106 expressions of dexamethasone group were lower than model group at 1,2,6 hour(p<0.05).The CD54 expressions of Sini decoction group were lower than model group at 1,2(p<0.01),3(p<0.05) hour.The CD56 expressions of Sini decoction group were higher than model group at 3(p<0.05),6(p<0.01) hour.The CD106 expressions of Sini decoction group were lower than model group at 1,2,3(p<0.01),6(p<0.05) hour.5.No expression of iNOS in normal rat brain.The expression of iNOS in model group was highest at 1 hour,decreased at 2,3 hour and rebounded at 6 hour.The iNOS expressions of dexamethasone group were lower than model group at 1(p<0.01),6(p<0.05) hour.The iNOS expression of Sini decoction group was lower than model group at 6 hour(p<0.01). Concentration of SOD in model group was lower than sham operated group on four time periods (p<0.01) and reached lowest valley at 1 hour.Concentration of SOD of dexamethasone group and Sini decoction group were higher than model group at 1,2 hour(p<0.01).Concentration of MDA in model group was higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.It's changes regularities are same as SOD.Concentration of MDA of dexamethasone group was lower than model group at 1,2,3,6 hour(p<0.01).Concentration of MDA of Sini decoction group was lower than model group at 2,3,6 hour(p<0.01).6.Electron microscopy and light microscopy showed:There was reactive injury at 1 hour, injury restoration at 2,3 hour,serious damage at 6 hour in endotoxic shock rat brain;rat brain damage of dexamethasone group reduced at 1 hour;rat brain damage of Sini decoction group reduced at 6 hour.TUNEL showed:There was a very small number of apoptotic cells in normal rat brain.Apoptotic cells of model group were less than sham operated group and reached peak at 6 hour.Apoptotic cells of dexamethasone group were less than model group atl,2 hour(p<0.01).Apoptotic cells of Sini decoction group were less than model group at2,3, 6hour(p<0.01).Conclusion1.There were 91 differentially expressed inflammatory cytokines in rat brain after endotoxic shock.There were 36 inflammatory cytokines involved with inflammatory response among them.TNF-α,IL-1β,IL-10 have been reported more and IL-1α,IL-13,IL-15,I15ra have been reported less.2.Organ damage induced by two ways in endotoxic shock:①Increasing prereceptor signal substances such as TNF-α,IL-1αinduced by LPS.→The expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,CD54.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.②The expression of cell-surface adhesion molecules,CD54,induced by LPS.→CD54 increased content of TNF-α,IL-1α.→Lead to inflammatory mediator waterfall cascade reactions and a large number of mediator was activated.→The expression of many cell-surface adhesion molecules,CD54,CD56,CD106.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.3.Sini Tang Injection has efficacy at treating shock and multifaceted roles.It can up-regulate blood pressure,SOD content,CD56 expression and down-regulate content of TNF-α, IL-1α,MDA and CD54,CD106,iNOS,apoptotic cells expression.Sini decoction took effect relatively late but lasting stability and dexamethasone relatively early but unstability.
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