Protective effects of chitosan oligosaccharide

1
Protective effects of chitosan oligosaccharide
on paraquat-induced nephrotoxicity in rats
Byung Chul Shin, Wan Soo Lee, Dae Wong Kang, Jong
Hoon Chung and Hyun Lee Kim Division of
Nephrology, Department of Internal Medicine,
College of Medicine, Chosun University
Results-2
Introduction

• Western blot analysis
• Western blot analysis of apurinic/apyrimidinic
  endonuclease (APE), p53-upregulated modulator of
  apoptosis (PUMA), caspase-3, and cleaved caspase-
  3 expression in the kidneys of paraquat
  (PQ)-intoxicated rats. The levels of these
  proteins increased in a time-dependent manner in
  the control group after PQ administration.
  Pretreatment with chitosan oligosaccharide (COS)
  resulted in a higher level of these proteins
  before PQ injection, which decreased at the end
  of experiment.
• Densitometric results in the control and chitosan
  oligosaccharide (COS) groups in
• paraquat (PQ)-intoxicated rats. Changes are
  compared bewteen groups at the same time point.
  p lt 0.05 p 0.05 Cas-3, caspase-3 Cl.
  Cas-3, cleaved caspase-3.
• Histology (PAS stain)

• Paraquat (PQ 1,1'-dimethyl-4,4'-bipyridinium
  dichloride)
• - world wide used herbicide and has a very high
  mortality rate in human
• Chitosans
• - N-deacylated derivative of chitin in crab and
  shrimp shells
• - Physiological activities antitumor
  activities, immuno-enhancing effect,
• ant-micorbial effect, cholesterol reducing
  effect and antioxidant effects .
• Apoptosis
• PUMA (p53 upregulated modulator of apoptosis)
  pro-apoptotic member of the Bcl-2 protein family
  and induction of programmed cell death6).
• APE (Apurinic/apyrimidinic endonuclease) repair
  of DNA damage and inhibits pro-apoptotic
  functions of P537).
• Caspse-3 important role in the apoptotic process
  in two ways8).
• First apoptotic gene PUMA and an effector of
  the caspase cascade, caspase-3, were evaluated to
  gain insight into apoptotic signaling during
  PQ-induced nephrotoxicity.
• Second, protective effects of COS were
  estimated through restoration of serological
  parameters and by PUMA and caspase-3 expression.
• Finally roles of apurinic/apyrimidinic
  endonuclease (APE) were also as-
• 103 sessed in PQ-induced nephrotoxicity.
• RatsMice
• Male SpragueDawley rats (810 weeks old, 200250
  g)

Purpose
Periodic acid Schiff (PAS) staining in the deep
renal cortex in paraquat (PQ)-intoxicated rats.
It clearly shows the proximal convoluted tubules
in both groups ((A) control group (C) chitosan
oligosaccharide (COS) group) before PQ injection
and also shows that the findings had disappeared
24 h after PQ administration (B). Only a few
proximal tubules bear brush borders (asterisks).
COS pretreatment prevented the degenerative
changes in proximal tubules (D), whereas the
distal tubules and collecting ducts were not
affected by PQ administration. Scale bar 50 um.
Methods
Immunohistochemistry with apurinic/apyrimidinic
endonuclease (APE) (upper column) and cleaved
caspase-3 (lower column) in the kidneys of
paraquat (PQ)-intoxicated rats. APE and cleaved
caspase-3 were mainly seen in the proximal
tubules of the control renal cortex. They were
found in whole components of nephrons from 4 h
after PQ injection, but degenerated proximal
tubules (asterisks) were devoid of
immunoreactivity 24 h after PQ injection.
Chitosan oligosaccharide (COS)-pretreated kidneys
showed ubiquitous distribution of immunolocalized
APE and cleaved caspase-3. The immunolocalization
did not change until 24 h after PQ injection,
even in the proximal convoluted tubules. G,
glomeruli. Scale bar 100 um.

Summary
Results-1
Conclusion