Refinement of LPS induced Sepsis in SD Rats to Mimic Human Sepsis
Meenakshi Bhardwaj, Rutuja Patil, Sugumar Mani, Malarvizhi R and A Hannah Rachel Vasanthi*
Natural Products Research Laboratory, Department of Biotechnology, School of Life Sciences, Pondicherry University, Puducherry, India 605014
Corresponding Author E-mail :hrvasanthi@gmail.com
Abstract: Systemic manifestations of infection due to bacteria or its toxins cause sepsis. Sepsis ultimately leads to multiple organ dysfunction syndrome and finally ends up in death and there are many experimental models to induce sepsis and study its prognosis. The present study aims to develop and standardize the effective concentration of Lipopolysaccharides (LPS) for induction of sepsis in male and female Sprague Dawley (SD) rat model as part of the refinement of animal protocols reducing pain and reduce stress in experimental rodents. Different concentrations of LPS (5, 10, 15 mg/ml) were administered to male and female rats (i.p.). Physical parameters like body temperature, heart rate and blood pressure along with hematological parameters (WBC, neutrophil, lymphocyte, platelet and hematocrit), oxidative stress markers (SOD, GSH and LPO), inflammatory markers (IL-2, TNF-α, NF-κB) and histopathology analysis were performed to confirm the induction of sepsis and its severity. Blood parameters like WBC, lymphocyte, platelet and hematocrit count significantly reduced whereas neutrophil count significantly increased in male SD rats in 10 and 15mg induction groups after 8h. LPS significantly increased generation of ROS, IL-2, TNF-α and NF-κB expression in the heart tissue at a dose of 10 and 15 mg in male SD rats than the female rats. Significant homogenous expression of blood, biochemical and molecular markers confirmed that 10 mg of LPS is sufficient to induce sepsis in experimental animals thereby reducing the manifestations and stress due to sepsis. However male rats are more suitable as compared to female rats, which might be resistant to such infectious toxins.
Keywords: Inflammation; Interleukin; LPS; Sepsis; TLR4 Back to TOC