Dissertation > Excellent graduate degree dissertation topics show

The Effect of Hemoperfustion in Different Time on the Proinflammatory Cytokines and Survival Time of Sepsis Rabbits

Author: JiGuoYe
Tutor: WangBin
School: Southern Medical University,
Course: Pediatrics
Keywords: Hemoperfusion Sepsis Rabbit Cytokines
CLC: R459.7
Type: Master's thesis
Year: 2011
Downloads: 19
Quote: 0
Read: Download Dissertation

Abstract


BackgroundSepsis is systemic inflammatory response syndrome caused by infection. It is confirmed that bacteria’s presence or highly suspicious lesions. Sepsis-severe sepsis-septic shock-multiple organ dysfunction-multiple organ failure-death is the final outcome of many patients. Currently, the pathogenesis of sepsis includes:host autoimmune injury, bacterial translocation and intestinal endotoxemia, inflammatory system and coagulation homeostasis, microcirculation and mitochondrial dysfunction syndrome. When pathogens invade the body, it will cause the immune responses. When the immune inflammatory response gets out of control, it can cause cytokine storm and inflammatory mediator falls and finally cause sepsis. Bacterial infection can be treated with antibiotics, but we can not restore the intestinal barrier function. The antibiotics kill the bacteria, also result in massive release of endotoxin. In this process, the secretion of pro-inflammatory cytokines stimulated by the interaction of endotoxin lipopolysaccharide (LPS) and host immune system is closely connected with the severity and prognosis of sepsis. Among these pro-inflammatory cytokines, the most important ones are TNF-α, IL-1, IL-6 and so on. TNF-α:As the origin of cytokines, its central role in the inflammatory response is to activate the cascade of cytokines, inducing the secretion of IL-1, IL-6, prostaglandins, which stimulate the inflammatory chain reaction. TNF-αcan cause fever, induce acute phase protein synthesis in liver cells, directly promote endothelial cell adhesion of neutrophils, monocytes, polymorphonuclear neutrophils (PMN) to go into sites of inflammation, release cytokines, participate and Zoom in SIRS, playing a leading role in the process of starting sepsis. IL-1:Synergizes TNF-αto start sepsis, causes fever, induces acute phase protein synthesis in liver cells, stimulates monocytes and macrophages to produce IL-6 and TNF, and mediates neutrophil chemotaxis by IL-8, which is produced by mononuclear cells and macrophage cells, amplifying the development of sepsis. IL-6:Not only can activate neutrophils, but also delay the phagocytes’ devourment of neutrophils which are anile and lose cell functions, thus exacerbate the inflammatory mediators, leading to immune dysfunction. Therefore, the blocking and reducing the pro-inflammatory cytokines, may become one of effective treatments of sepsis.Blood purification refers to the patient’s blood is drawn out in vitro and some pathogenic substances are removed through a cleaning device, the blood is purified, so as to cure diseases. This process is the blood purification. Blood perfusion (hemoperfusiom, HP) therapy is to make the patients’ blood flow through the pipeline toward hemoperfutors, and the blood will be purified or treated biologically by the adsorbent or other biological materials when it passes through the hemoperfutors. Then after hemoperfusion, the blood returns to veins through the pipeline. In the HA Resin hemoperfutors, the adsorbent is a styrene/divinylbenzene macroporous polymer, it is a microporous resin-by adjusting the reaction temperature, stirring speed, and the use of polymer suspension polymerization methods, it controls cross-linking and chooses resin porogen to adjust aperture to a specific range, selects the cross-linkers which are rich in lipophilic hydrophobic groups to be cross-linked, so that the resin adsorbent become lipophilic hydrophobic adsorbent, properly regulates the coated liquid system boiling point and adjusts the pore size of drying temperature; then it is accomplished by selecting concentration of coated liquid and dries speed control film thickness. It is biocompatible, and the major adsorption mechanism is:It directly absorbs and clears the correlative pathogenic material of blood by the neutral macroporous adsorption resin as adsorbent, absorbs physically and interacts among hydrophobic groups using the resin molecular sieve effect of three-dimensional network structure. It is characterized by the adsorption capacity, fast adsorption, high mechanical strength, relatively specific adsorption, can effectively remove the pathological elements in blood, especially for relatively specific adsorption capacity with the pathological molecules combining mid-large molecules and proteins. So it has currently been widely used in clinical treatment of patients with sepsis. However, when is the best time to be treated with hemoperfusion in sepsis? Currently there is no definitive answer at home and abroad. We assume that we make the rabbit sepsis model by cecal ligation and puncture surgery, then according to the time after cecal ligation and puncture,the rabbits are divided into different groups, and respectively treated with hemoperfusion once, while we test the blood concentrations of pro-inflammatory cytokines TNF-α, IL-1β, IL-6 which are are closely relative with sepsis in multiple time points, compare the changes, find out the best timing for sepsis rabbits treated with hemoperfusion by combining with the survival time which is closely relative with the prognosis of sepsis, so as to provide an animal experimental basis for the choice of hemoperfusion time of clinical sepsis patients, and to improve the prognosis of sepsis patients.Objective:To investigate the best timing to be treated with hemoperfusion for sepsis rabbits by detecting the concentrations of TNF-α, IL-1, IL-6 and the rabbits’ survival time when the rabbits are teated with hemoperfusion in different time.Method:1、2 male New Zealand white rabbits were randomly divided into 7 groups,6 in each group. After the intravenous anesthesia with sodium pentobarbital, each rabbit is undergo cecal ligation and puncture (CLP)surgery to be made sepsis model, and femoral artery and vein catheter is performed in need of providing a blood line channels during hemoperfusion.2、ach rabbit was treated with hemoperfusion once respectively in 3 hours,6 hours,9 hours,12 hours,24 hours,36 hours after the CLP with a self-made hemoperfutor. Perfusion time was 2 hours, speed 5ml/min.Empty perfusion group rabbits were treated with empty hemoperfusion (perfused with air perfutors) in 3 hours after CLP as the experimental control group.3、ach rabbit was taken peripheral blood samples 2mL before CLP, after 3 hours,6 hours,9 hours,12 hours,24 hours,36 hours,48 hours (8 time points in all) then the serum samples were tested together with ELISA kits. After the completion of reaction in each plate hole, added the stop solution and measured the OD values at 450nm wavelength with a microplate reader. We used the OD values of the standard holes to obtained the regression equations by SPSS software, input the OD values of each sample into the equations, then we obtained the blood samples’ concentrations of TNF-α,IL-1βand IL-6.4、Rabbits were fixed on the operating table until death, and regularly noted the spirit, breathing, heart rate, body temperature of rabbits. Daily provided intravenous fluid, and the fluid volume was 80ml/kg, including 0.9% natural saline and 10% glucose solution, the total tension was 1/4. At last we recorded the survival time.5、We used the SPSS13.0 statistical software. The concentrations of IL-6, IL-1βand TNF-αwere analysed with two-factors multi-levels repeated measures analysis of variance. Hemoperfusion was the intervention factor (7 group), and there were 8 levels in the measurement time factors (8 measurement points). Then we analysed the linear correlations of the three indexes. We analysed the survival time with Kaplan-Meier method, and mapped the survival curves. The test was set atα=0.05.Results:1、The results of the observed indexes:42 rabbits began to suffer from mental fatigue, gradually growing fast of breathing, heart rate and other symptoms in about 12 hours after CLP. All died in 48 hours to 90 hours after CLP. Rabbits were anatomized after death, in the abdominal cavities there were purulent discharge on the surface of intestine and peritoneum, purulent or bloody ascites, bad smell. Under the ligation site, the bowel was black and purple, crisp, and it adhered with the surrounding bowel and peritoneum. Survival time of rabbits was analysed with survival analysis (Kaplan-Meier method). The result showed that each perfusion group were generally significantly different (Chi-Square:57.313, P<0.001); Pairwise comparison,12h perfusion group had the longest survival time (78.667+8.262 hours).2、The results of the detection indexes:(1) TNF-α:Repeated measures analysis of variance, test of sphericity:W 0.000, P<0.001, does not meet the spherical assumption, Greenhouse-Geisser correction factor is:0.265. Main effects:there were significantly difference among different perfusion groups overall, F value 18.231, P<0.001; there were significantly difference among the repeated measurement factors of 8 different time points overall, F value 179.566, P<0.001. Pairwise comparison among perfusion groups (LSD method):there were significantly difference, the average concentration of TNF-a in the perfusion group after CLP 3h was the lowest (80.750pg/ml); Pairwise comparison among time points:there were significantly difference, the average concentration of TNF-αin the time point after CLP 6h was the highest (937.287 pg/ml). Interactive effects and individual effects:time of repeated measurements interacting with different levels of perfusion group:F value 5.531, P<0.001, there was interaction between time of repeated measurements and different perfusion groups. When the air perfusion group was at the time which is after CLP 6h, the average concentration of TNF-a was the highest.(2) IL-1β:Repeated measures analysis of variance:test of sphericit:W=0.000, P<0.001, does not meet the spherical assumption, Greenhouse-Geisser correction factor is:0.150. Main effects:there were significantly difference among the perfusion groups overall, F=8.391, P<0.001; there were significantly difference among the repeated measurement factors of 8 different time points overall, F=96.566, P<0.001. Pairwise comparison among perfusion groups (LSD method):there were significantly difference, the average concentration of IL-1βin the perfusion group after CLP 6h was the lowest (36.160 pg/ml); Pairwise comparison among time points:there were significantly difference, the average concentration of IL-1βin the time point after CLP 9h was the highest (265.251 pg/ml). Interactive effects and individual effects: time of repeated measurements interacting with different levels of perfusion group:F value 4.328, P=0.002, there was interaction between time of repeated measurements and different perfusion groups. When 24h perfusion group was at the time which is after CLP 9h, the average concentration of IL-1βwas the highest.(3) IL-6:Repeated measures analysis of variance:test of sphericit:W=0.000, P <0.001, does not meet the spherical assumption, Greenhouse-Geisser correction factor is:0.162. Main effects:there were significantly difference among the perfusion groups overall, F=2.394, P=0.048; there were significantly difference among the repeated measurement factors of 8 different time points overall, F value 91.880, P <0.001. Pairwise comparison among perfusion groups (LSD method):there were significantly difference, the average concentration of IL-6 in the perfusion group after CLP 12h was the lowest (521.957 pg/ml); Pairwise comparison among time points: there were significantly difference, the average concentration of IL-6 in the time point after CLP 12h was the highest (4492.715 pg/ml). Interaction:time of repeated measurements with different levels of perfusion group interaction:F value 0.903, P= 0.512, there was no interaction between time of repeated measurements and different perfusion groups.Conclusion:Hemoperfusion in the early and medium-term process(after CLP 3h to 12h) of sepsis can most effectively reduce the blood concentration of Pro-inflammatory cytokines, and hemoperfusin in the medium-term process(after CLP 12h) of sepsis, can also make the survival time of rabbits longest. It indicates that the pro-inflammatory cytokines of the clinical patients with sepsis may be effectively reduced when the patients were treated with hemoperfusion in the early and medium-term process of sepsis, and the prognosis may be improved.

Related Dissertations

  1. MRI Guoded Experimental Cryoablation of Rabbit’s Sciatic Nerve for MRI and Pathology Contrast Study,R445.2
  2. The risk Beihai academic thinking and clinical experience and the application of gastrointestinal recovery method of therapeutic effects of sepsis, intestinal dysfunction,R249.2
  3. Multi-center Study of Early Hemofiltration Treat Sepsis Associated AKI,R692.5
  4. The Polymorphisms of TLR4 Gene and Function Analysis of C1027A in Suzhong Pig,S828
  5. Nutritional Evaluation and Feed Effects of Solidago Canadensis in Rex Rabbits,S829.1
  6. The Study on Influence of Transcriptional Profiles of Porcine Peripheral Blood Lymphocytes Cultures Following Infection with Porcine Parvovirus,S858.28
  7. The Study of Rabbit Bone Marrow Mesenchymal Stem Cells Separation Culture and Directional Differentiation,R329
  8. Establishment of Feline Calicivirus (FCV) Reverse Genetics System Using RNA Polymerase Ⅱ and Expression of RHDV VP60 Gene,S852.65
  9. Effects of the Applied External Weak Steady Direct Current Electrical Fields on Diastolic Function after Balloon Injury in the Rabbit Abdominal Aorta,R541.4
  10. Clinical Efficacy Observation with Fu-Zheng-Gu-Ben on Septic Acute Deficiency Syndrome Patients,R278
  11. Effects of Geniposide and Its Metabolite Genipin on Human Peripheral Blood T Lymphocytes,R758.23
  12. Effect of Sciatic Injection Injury on Enzymohistochemistry of Skeletal Muscle in Rabbit’s Legs,R-332
  13. Effects on Recombinant Human Interleukin-10 of IL-2, IFN-γ, GCs, T Cell Apoptosis in Transplantation of the Rabbit Skin,R392
  14. The Study of 3D Co-culture with Fibroblast and Gelatin Sponges,R329
  15. Septic lung injury (ALI / ARDS) in patients with peripheral immune parameters Th1, Th2, HLA-DR and Treg changes and Integrative Medicine,R459.7
  16. The Significance of Triggering Receptor Expressed on Myeloid Cells-1(TREM-1) in Neonatal Rats Septicemia with E.coli,R363
  17. The Effect of Different Concentration of Urokinase on Stereotactic Aspiration of the Rabbit Intracerebral Hematoma and the Expression of the Heme Oxygenase-1,R743.34
  18. GDF-5 and Rabbit Disc in Vitro Model Induction BMSCs of Rabbit Differentiation to the Nucleus Pulposus Cell,R329
  19. Subtalar osteotomy for treatment of old fractures slowly extended research,R687.3
  20. Pentamethylquercetin on diabetic nephropathy in rats and its mechanism,R285.5

CLC: > Medicine, health > Clinical > Therapy > Emergency, first aid
© 2012 www.DissertationTopic.Net  Mobile