On the other hand, in patients with primary liver injury, such as viral hepatitis, LPS plays an important role in intestinal endotoxemia, which in turn induces secondary liver injury and liver failure[1]

On the other hand, in patients with primary liver injury, such as viral hepatitis, LPS plays an important role in intestinal endotoxemia, which in turn induces secondary liver injury and liver failure[1]. systemic vascular resistance index (SVRI) and left ventricular work index (LVWI) of macaques were significant declined in shock model group on average 60 min after LPS injection. The plasma levels of TNF- and IL-10 were significantly increased 60 min after LPS injection and then decreased. The plasma levels of IL-1 and IL-12P40 were significantly increased at 120 min after LPS injection. The mRNA levels of TNF- and IL-1 were significantly increased Benzocaine 60 min after LPS stimulation in PBMCs and 120 min after LPS stimulation in livers. The mRNA level of IL-18 was significantly increased 120 min after LPS stimulation in PBMCs and livers. But in Benzocaine spleen, only TNF- mRNA level in LPS group was significantly higher 120 min after LPS stimulation, compared with that in control group. CONCLUSION: An endotoxic shock model of Macaque mulatta was successfully established. Both antibodies for ELISA and PCR primers based on human cytokine assays were successfully applied to detect macaque cytokines. In the model, inflammatory cytokines, such as TNF- , IL-1 , IL-12 and IL-18 as well as anti-inflammation cytokine IL-10, were released at very early phase of endotoxic shock within 120 min after LPS injection. PBMCs and liver cells might be the important sources of these cytokines. INTRODUCTION Endotoxin-induced shock mostly occurs during serious infection with Gram-negative bacteria. Lipopolysaccharide (LPS), residing in the outer membrane of all Gram-negative bacteria and being the major component of Gram-negative bacteria cell walls and the toxic component of endotoxin, is considered as an important initiating factor of the Gram-negative septic syndrome in human. Gram-negative sepsis has aroused great concern in clinic because of its high mortality. But LPS is not the direct causative factor. It is well known that LPS activates immunocytes, such as monocyte-macrophage and lymphocyte, and induces these cells to excessively release a series of potent inflammatory cytokines, including TNF- , IL-1 , IL-6, IL-8 and IL-18. It is the excessive cytokines, such as TNF- , IL-1 , IL-6 and IL-18, that result in high fever, hypotension, vascular endothelial cell damage and disseminated intravascular coagulation, blood capillary leak syndrome, and multiple organ failure. On the other hand, in patients Benzocaine with primary liver injury, such as viral hepatitis, LPS plays an important role in intestinal endotoxemia, which in turn induces secondary liver injury and liver failure[1]. New therapeutic concepts for the treatment of endotoxic shock or endotoxemia with anti-inflammatory cytokines have been developed. Although knowledge about the changes of inflammation-associated cytokines during endotoxemia and endotoxic shock has been well addressed, the change pattern of inflammatory cytokines just during the early phase of endotoxemia and endotoxic shock is still not clear and worth working on, because it can help the selection of new therapeutic targets. The therapies in early phase are more effective. The aim of the present study was to investigate the changes of TNF- , IL-1 IL-10, IL-12 and IL-18 expressions during early phase (within 120 min) of endotoxic shock in macaques. MATERIALS AND METHODS Animals Totally 25 macaques (O127: B8 and prepared by phenol extraction, was purchased from Sigma Co. (Saint Louis, USA). ELISA kits for detecting human TNF- , IL-1 , IL-10 and IL-12 P40/ P70 were from Pharmingen (San Diego, USA). TriZol RNA extraction kit was from Gibco (Grand Island, USA). Moloney murine leukemia virus (M-MLV) reverse transcriptase, RNase inhibitor, PCR kit and PGEM-T vector were all from Promega (Shanghai, China). PCR product purification kit was from Boehringer Mannheim Co. (Mannheim, Rabbit Polyclonal to CDC25C (phospho-Ser198) Germany). Plasmid DNA extraction kit was from Scientz Bio Co. (Shanghai, China). PCR primers In order to determine the mRNA expression levels of inflammatory cytokines by RT-PCR, some pairs of primers based on specific sequences published previously were used. These primers, synthesized in ShengGong Biotech Co. (Shanghai, China), are shown in Table ?Table11. Table 1 PCR primer sequences for TNF-, IL-1 and IL-18 detection .