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Ethyl pyruvate act as an antioxidant and anti-inflammatory

drug against sepsis induced acute kidney injury.(Reade and

Fink,2005)

Chitinase-like proteins are candidate biomarkers for sepsis-

induced acute kidney injury(Maddens et al.,2012)

Animal models of sepsis and sepsis-induced acute kidney

injury (Doi et al.,2016)

Recent updates

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Ethyl pyruvate shows protective action against sepsis (LPS and

CLP) induced acute kidney failure in aged mice, by down-

regulating Serum TNF-α, Cyr61 expression, mRNA abundance

for TNF-α, PAI-I, tPA and by up-regulating mRNA abundance for

uPA. Ethyl pyruvate protect against multiple organ damage cause by

CLP model of sepsis. Volume resuscitation in LPS model and that of with antibiotic

treatment in CLP model prevent acute renal failure.

Hypothesis

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1) Primary objectives:- To investigate effect of ethyl pyruvate on LPS and CLP induced acute renal failure by sepsis.

To check out either Volume resuscitation in LPS model and that of

with antibiotic treatment in CLP model prevent acute renal failure or

not.

2) Secondary objective:- To check out either Ethyl pyruvate protect against multiple organ

damage cause by CLP model of sepsis or not.

Objectives

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The mortality rate of ARF in septic patients is 75%, and 45% in patients

without sepsis.(NEVEU et al., 1996)

Lipopolysaccharide(LPS)-induced systemic inflammation and cecal

ligation puncture (CLP)-induced polymicrobial infection models are

used commonly to screen drugs and study the pathogenesis of

sepsis. (Neild et al., 2001)

Ethyl pyruvate scavenges reactive oxygen species (ROS), down-

regulates proinflammatory cytokines, inhibits high mobility group

box1 protein (HMGB1), a late mediator of sepsis, and decreases the

activation of p38 and NF-kβ. (ULLOA et al., 2002, WANG et al., 1999, YANG et al.,

2002)

Introduction

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Drug Profile

ETHYL PYRUVATE

Antiinflammatory

Antioxidant Anticancer Cardio protective

Neuroprotective

(Vyawahare et al., 2012)

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Animal:- Young (7 to 8 weeks) and aged (42 to 44 weeks) male C57BL/6 mice were use in study with total 8 groups as,

Experimental Groups

Normal group ( vehicle solution, N=4 to 10)

W/O F, LPS induced(o.o8 mg/kg, iv, N=4 to 10)

W/F(IA), LPS induced(o.o8 mg/kg, iv, N=4 to 10)

W/O F, LPS induced(o.o8 mg/kg, iv, N=4 to 10)

CLP surgery (Diseased induced, N=15)

W/F(IA) and antibiotics (CLP induced, N=4 to 10)

Sham injected (CLP induced, N=10)

RL or Ethyl pyruvate(8 or 40 mg/kg, IP), (CLP induced, N=4 to 10)

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Experimental ProtocolLPSCLP

Animals received 0.4 mL of Ringer’s lactate or a similar volume of freshly made Ringer’s ethyl pyruvate (8 or 40 mg/kg, ip). A single dose was injected at 0, 6, or 12 hours after CLP surgery

1.5 mL of 3/4 normal saline was given at 0, 6, or 18 hours after LPS injection, and then every 12 hours.

Volume and antibiotic treatment

every 12 hours, starting at 6 hours after surgery

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Performed Parameters

Survival study

Blood chemistries and serum TNFα measurements

Histological examination

Western blot analysis for Cyr61

Measurement of mRNA abundance by RTPCR

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Creatinine and Survival study

Volume resuscitation in LPS induced model, W/Fluid in aged (0,6,18hrs) and W/O

Fluid in young mice significantly reduces PA-Cr

serum level.

Volume resuscitation and antibiotics treatment in CLP

model, body weight increased by 7%, and the survival was 100% at 24

hours, 43% at 48 hours, and 14% at 72 hours.

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Characterization of the volume and antibiotic-treated

CLP model in aged mice

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CLP sepsis-induced ARF in volume and antibiotic-treated

aged mice

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Time course of renal histologic changes. Kidney sections were stained with either periodic acid-Schiff (AtoD) or naphthol AS-D chloroacetate esterase (E). Typical histology in sham-treated animals at 24 hours (A) or cecal ligation puncture (CLP) animals at 6 hours (B) and 24 hours (C) are shown. Typical histology in animals treated with ethyl pyruvate at 24 hours (D). Arrows, esterase positive cells (original magnification,250 (A to D),400 (E). (F) Time course of the tubular damage. The tubular damage score was measured in the cortex (right panel) or the outer stripe of the outer medulla (OSOM) (left panel). Values are mean SE (N=5 to 6 per group). *P0.05 vs. sham. (G) Time course of renal Cyr61 expression. Renal Cyr61 measured at indicated times (hours) after CLP. N, normal; S, sham; and P, positive control for Cyr61

Histopathological study

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Effect of CLP sepsis on renal Cyr61 expression and serum TNF-α

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Effects of ethyl pyruvate on CLP-induced multiple organ damage and ARF

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Effects of ethyl pyruvate on ARF following CLP sepsis.

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Effects of ethyl pyruvate on mediators after ARF following CLP sepsis.

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LPS/CLP

Proinflammatory cytokines

In-vivo TNF-α In-vitro

TNF-α

HMGβ1

P38(MAPK)

NF-kβ

mRNA for TNF-

α

Action on coagulation

Decrease mRNA abundance for uPA, Increase tissue factor

and PAI-I mRNA

ETHYL PYRUVAT

E

Mechanisms of action

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Discussion Model of sepsis-induced ARF that mimics the

human disease:- Study developed a new model of sepsis-induced ARF based upon the

polymicrobial CLP model, but with two distinctive features: (1) appropriate

volume and antibiotic resuscitation, and (2) use of aged mice. volume resuscitation schedulethat inhibited LPS-induced renal damage

and increased body weight by 7% to mimic the fluid overload often

present in hypotensive intensive care unit patients. Study used aged mice because they more closely match the age

distribution of sepsis-induced ARF and because the elderly are more

prone to sepsis-induced ARF.

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Effects of ethyl pyruvate on sepsis-induced ARF:-

Ethyl pyruvate has been shown to decrease mortality in both the LPS and

CLP models and is effective even when given 24 hours after CLP sepsis

Ethyl pyruvate still protected against renal and muscle injury even when

treatment is started 12 hours after surgery.

Thus, ethyl pyruvate may be a treatment for both sepsis and sepsis-

induced renal and multi organ injury. This prolonged window of

opportunity may be important clinically because of the difficulty in the

early detection of sepsis and sepsis-induced ARF.

Continue…

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volume- and antibiotic treated aged mouse model of sepsis mimics

many aspects of human sepsis. This model should be useful for screening therapeutic agents and

determining mechanisms of disease pathogenesis and protection.-

induced ARF that Ethyl pyruvate inhibits sepsis-induced renal and multiorgan damage,

even when started 12 hours after surgery. by inhibiting dysregulated

inflammatory and coagulation/hemostatic pathways Ethyl pyruvate may hold promise in clinical settings, since it can

prevent the initiation and protect the progressive phase of sepsis-

induced ARF.

Conclusion

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