| Background :From animal experiment, the rats were handled by lethal dose lipoplysaccharide (LPS); The early inflammation factors as TNF, IL-1b etc. General reached the maximizing in 6h, after that these factors quickly went down to basic level, but the rats still were death after a few days attacked. This phenomenon pointed out one later transmitter was in the course of death by sepsis. After 18h we fund a sort of protein appeared in RAW264.7 cell of the rat stimulated by lipoplysaccharide (LPS), the molecular weight of this protein was 30 Kilodalton, but this protein was inapparent in early part. Analysis approved this late transmitter was high mobility group protein B 1 (HMGB1) of the rat. HMGB1 was one of the high mobility group (HMG), the HMG included HMGB, HMGN, HMGA, the HMG B1 had two"L"type HMG box made up of about 80 amino acid, both of them were A box (1-85aa) and B box (88-162aa), and the field combined DNA. The B box was the main field of the HMGB1 induced inflammation, the A box could emulatively restrain the function of the HMGB1 induced inflammation. Macrophages, monocyte formed rich HMGB1 vesica in cell after the stimulation by IL-1, TNF, these cell released HMGB1 by exocytosis; At one time, macrophages, monocyte released inflammation factors under the stimulation by HMGB1, as TNF, IL-1α, IL-1b, IL-6, IL-8 etc. The tumor necrosis factor alpha (TNF-α) was a sort of broad biological active cell factors, TNF-αwas basically brought by enabled macrophages and lymphocyte. Such as body released TNF-αby the stimulation of endotoxin, moreover the release of TNF-αcould arouse the release of else factors as IL-1, CSF etc. All these factors had important action in the startup, keep, fadeaway of inflammation. IL-10 was a kind of important inhibiting inflammation factors, also was named the factor of inhibiting the cell factor composition, macrophages and monocyte stimulated inhibiting the release of inflammation factors and advancing the release of cell B and the exudation of immunity globin were the primary biological characteristic of this factor, and this factor had immunity inhibition in the function of macrophages. Three factors HMGB1, TNF-α, IL-10 had some dissimilarity in each biological characteristic, and brought into play different effect in the appearance, development, change of sepsis, at the some time the affiliation was among them. Clarifing the correlation among the three factors would offer usable clew and information in the basic study and clinic therapy of sepsis. Objective :Our study object was the patients of acute obstructive suppurative cholangitis, mensurating the gene and proteinic expression of HMGB1, TNF-α, IL-10 in the blood of pyemic patients, and discussing the expression of the HMGB1 in the blood of pyemic patien, and analyzing its clinic significance, offering the academic basis for the clinic therapy of sepsis.Methods :Selecting the case under the definiens and diagnostic standard of sepsis, our cases were choosed by dianonist criteria derminationed in international sepsis conference in 2003. Sepsis is the systemic phlogistic reaction owing to infection, body had obvious changes in temperature, circulation, breath. It was systemic reaction of body to infection, had two conditions of Systemic inflammatory response syndrome (SIRS) (SIRS+evidence of infection). SIRS consisted with the following two or upwards two phenomenon: T>38℃or<36℃;P>90/min; R>20/min; PaCO2<4.27 kPa; WBC>12×109/L or<4×109/L, or immature granulose cell>10%, acute obstructive suppurative cholangitis (AOSC) is the representative, the AOSC diagnostic criteria in our study was determination by surgery published in 2003: Reynolds'five signs: (1) bellyache, (2) chill and hot, (3) jaundice, (4) shock, (5) restrained phenomena of ganglia systerm; Laboratory test: (1) hoist of leucocyte count and neuter granulose cell, (2) fall of blood platelet count, (3) delay of thrombin time etc;Image examine; Cholangiolitis'three signs: (1) bellyache, (2) chill and hot, (3) jaundice and T>39℃, P>120/min, WBC>20×109/L, fall of blood platelet count. The cases fulfill all above conditions could be brought into the range of study, 23 cases accorded with those condtions. The healthy donors were 23 cases.1. The difference of the HMGB1 B box gene transcription level: The peripheral vein blood of AOSC patients and healthy postulants were taken preoperatively and 12, 24, 36h postoperatively. Total RNA of peripheral blood mononuclear cells(PBMC) were extracted and HMGB1 mRNA were amplified by RT-PCR to detect the expression of HMGB1 mRNA in PBMC of AOSC patients.2. The difference of the HMGB1 B box gene express level: The peripheral vein blood of AOSC patients and healthy donors were taken preoperatively and 12, 24, 36h postoperatively. The blood serum were extracted and the content of HMGB1 protein in blood serum was detected by western blot.3. TNF-αand IL-10: The contents of TNF-αand IL-10 proteins in blood serum were determined by ELISA.4.The three results were analyzed with SPSS12.0 Stat. software.Results:RT-PCR results showed that the express of HMGB1 mRNA in PBMC of AOSC patients was 0.82±0.04 raised up obviously and was significantly higher than that of healthy donors was 0.31±0.06 (P<0.01) and was decreased to normal level gradually after treatment biliary passage drainage, the 12, 24h postoperative were 0.46±0.05 and 0.38±0.07 (P<0.01).Western blot results showed that The content of HMGB1 protein in blood serum of AOSC patients was 0.81±0.03 was also significantly higher than that of healthy donors was 0.35±0.05 (P<0.01). And the content of HMGB1 protein in blood serum of AOSC patients decreased gradually after biliary passage drainage, the 12, 24h postoperative were 0.56±0.05 and 0.36±0.07 (P<0.01).ELISA results showed that the contents of TNF-αand IL-10 proteins of blood serum were 469±35pg/ml and 33.369±6.443pg/ml were significantly higher than the contents of healthy donors were 102±16pg/ml and 11.490±1.088pg/ml (P<0.01) and decreased gradually after biliary passage drainage, the 12h postoperative were 243±26 pg/ml and 20.562±5.630 pg/ml (P<0.01).Conclusions :(1) The content of HMGB1 protein and the express of HMGB1 mRNA in the peripheral vein blood of AOSC patients were significantly higher than those of healthy donors; (2) With pathogengtic condition's improvement after biliary passage drainage, the content of HMGB1 protein and the express of HMGB1 mRNA decreased gradually to normal level; (3) The HMGB1 was one level of significnance could reflect the severe and danger degree of pathogenetic condition in sepsis patients. |
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