Anvendelse af dyremodel af Sepsis at evaluere Nye Naturlægemidler Terapier
Summary
Sepsis henviser til en systemisk inflammatorisk respons syndrom skyldes en mikrobiel infektion, og kan simuleres ved en kirurgisk teknik kaldet cecal ligering og punktering (CLP). Her beskriver vi en fremgangsmåde til at anvende CLP-induceret dyremodel til screening lægeurter for terapeutiske midler.
Abstract
Sepsis henviser til en systemisk inflammatorisk respons syndrom skyldes en mikrobiel infektion. Det har været rutinemæssigt simuleret i dyr ved adskillige teknikker, herunder infusion af eksogent bakterielt toksin (endotoksæmi) eller bakterier (bakteriæmi), samt kirurgisk perforering af coecum ved cecal ligering og punktering (CLP) 1-3. CLP kan bakterierne spild og fækalt kontaminering af den peritoneale kavitet, efterligne den humane kliniske sygdom perforeret blindtarmsbetændelse eller diverticulitis. Sværhedsgraden af sepsis, som afspejlet i de endelige dødelighed, kan styres kirurgisk ved at variere størrelsen af nålen, der anvendes til cecal punktering 2. Hos dyr, inducerer CLP lignende, bifasisk hæmodynamisk kardiovaskulære, metaboliske, og immunologiske reaktioner som observeret under kliniske forløb af menneskets sepsis 3. Således er CLP-modellen betragtes som en af de mest klinisk relevante modeller for eksperimentel sepsis 1-3. </ P>
Forskellige dyremodeller er blevet anvendt til at belyse de indviklede mekanismerne bag patogenesen af eksperimentel sepsis. Den letale konsekvens af sepsis skyldes dels en overdreven ophobning af tidlige cytokiner (såsom TNF, IL-1 og IFN-γ) 4-6 og sene proinflammatoriske mediatorer (f.eks HMGB1) 7. Sammenlignet med tidligere proinflammatoriske cytokiner, har sen virkende mediatorer et bredere terapeutisk vindue til kliniske anvendelser. For eksempel, forsinket indgift af HMGB1-neutraliserende antistoffer begyndende 24 timer efter CLP stadig reddet mus fra dødelighed 8,9, om HMGB1 som et sent mediator for letal sepsis. Opdagelsen af HMGB1 som en sen-virkende mægler har indledt et nyt felt for undersøgelse for udvikling af sepsis behandlinger ved hjælp af traditionel kinesisk urtemedicin. I dette papir, beskriver vi en procedure for CLP-induceret sepsis, og dens anvendelse i screening urtemedicin for HMGB1 målretning terapier.
Protocol
Discussion
I laboratoriet har flere dyremodeller af sepsis blevet anvendt til at forstå patogenesen af sepsis for at udvikle potentielle nye terapier. Deres kliniske relevans er stadig genstand for debat, før den vellykkede oversættelse af dyreforsøg i kliniske ansøgninger om sepsis. Selv neutraliserende antistoffer mod tidlige cytokiner (f.eks TNF) var beskyttende i dyremodeller for bakteriæmi / endotoksæmi 17,18, de faktisk forværre overlevelse dyremodel af sepsis 19. På samme måde, de flest…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
Dette arbejde blev støttet af tilskud fra National Institutes of Health, National Institute of General Medical Science (R01GM063075) og National Center for Komplementær og Alternativ Medicin (R01AT05076).
Materials
| Name of the reagent | Company | Catalogue number |
| Betadine | Purdue Products L.P. | 25655-41-8 |
| imipenem | Merck & Co., Inc. | 9882821 |
| Ketamine HCl | Hospira Inc. | RL-0065 |
| Xylazine | Lloyd Laboratories | 4821 |
| Autoclip | Becton Dickinson | 427631 |
| 4-0 silk suture | Roboz | SUT-15-2 |
| Surflo I.V. Catheter | Terumo | SR*OX2419CA |
| RayBio mouse cytokine antibody array | RayBiotech, Inc. | AAM-CYT-3 |
| Thioglycollate | Becton Dickinson | 211716 |
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