Effects of Furosemide on Lipopolysaccharide-induced Disseminated Intravascular Coagulation in Rats

• Main Authors: Hao-Yuan Hung, 洪浩淵
• Other Authors: Chin-Chen Wu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/64635626969491825362

碩士 === 國防醫學院 === 藥理學研究所 === 101 === Disseminated intravascular coagulation (DIC) is a common complication of sepsis and leads to multiple organ failure (MOF) and death. Recent evidence suggests that oxidant stress and cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha play a major role in coagulopathy of sepsis. Lipopolysacchride (LPS) of Gram-negative bacilli induces the expression of tissue factor on monocytes and endothelium, which results in coagulation activation and microvascular thrombosis. In addition, the release of cytokines (e.g. TNF-alpha) stimulated by LPS impairs the anticoagulation system and fibrinolysis, which further promotes intravascular fibrin deposition and leads to DIC and MOF ultimately. A growing evidence manifests that furosemide is a powerful anti-oxidant and anti-inflammatory reagent in addition to its diuretic effect. Therefore, this study tried to evaluate the effects of furosemide on LPS-induced DIC defined by ISTH score system in Wistar rats. Wistar rats were divided into four groups: (1) Control, (2) Control + furosemide (3 mg/kg for 10 min i.v. infusion at 1 h), (3) LPS (10 mg/kg for 40 min i.v. infusion) and (4) LPS + furosemide (3 mg/kg for 10 min i.v. infusion at 1 hr after LPS). Whole blood sample was collected at 0, 1, 2, 4 and 6 h. The changes of hemodynamics, blood glucose, ISTH scoring parameters (i.e. platelet, prothrombin time (PT), fibrinogen (FIB) and D-dimer), cytokines (TNF-alpha and IL-6), thrombin-antithrombin complex (TAT), nitric oxide (NO), lactate dehydrogenase (LDH), hepatic (ALT) function, renal (BUN and CRE) function, and survival rate were monitored over 6 h. The vital organs including aorta, lung, liver and kidney were harvested at 6 h after LPS challenge to perform Western blot (expression levels of inducible nitric oxide synthase (iNOS), Na+-K+-2Cl− cotransporter 1 (NKCC1) and plasminogen activator inhibitor 1 (PAI-1)) and histo-pathological studies. The survival rate in each group was calculated at the end of studies. Data showed that LPS group matched the diagnostic crriteria of the ISTH for overt-DIC. The administration of furosemide increased the concentration of FIB and platelet count, decreased the level of plasma D-dimer, TAT, LDH, ALT, BUN and CRE, improved prolongation of PT and LPS-induced delayed hypotension, and increased survival rate in LPS-induced DIC rats. In addition, furosemide not only attenuated the iNOS, NKCC1 and PAI-1 expression in the lung and liver, but also attenuated the superoxide in the aorta, lung and kidney. Moreover, furosemide had therapeutic effect by reducing serum TNF-alpha, IL-6 and NO levels and organ superoxide levels, preventing coagulopathy of endotoxemia. Thus, this study reveals that furosemide may have beneficial effects on LPS-induced DIC and prevention of MOF in endotoxemia.