1. Protective effect of Piper betle leaf extract against cadmium-induced oxidative stress and hepatic dysfunction in rats
King Saud University Saudi Journal of Biological Sciences

2. Introduction Exposure Agricultural soils Mining activities of mines
Contaminated plant sources
Human activities
Global cycle of heavy metals & metalloids
Toxic non-essensial element
Mining activities of mines
Exposure
Accumulates in biological system
Injury to organs
Disfuction organs

3. Ethnomedicines in India
Bioactive molecules
polyphenol
Antioxidative property
alkaloids
steroids
saponin
tannins
Piper betle
Piper betle
Ethnomedicines in India
Low cost
Easy accesibility to everyone
Perceived fewer side effects
The possible hepatoprotective activity of Piper betle leaves against Cd induced hepatotoxicity has not been reported so far

4. Objective
To evaluate the hepatoprotective effect of the P.betle leaves by using the Cd-induce sub-chronic oxidative liver injury model in rats

5. Materials and methods

6. 2-thiobarbituric acid (TBA) Butylated hydroxytoluene (BHT)
Chemicals
Cadmium chloride
2-thiobarbituric acid (TBA)
Butylated hydroxytoluene (BHT)
Reduced glutathione (GSH)
2,2-dipyridyl
Xylenol orange
2,4-dinitrophenylhydrazine (DNPH)
γ-glutamyl-p-nitroanilide
5,5-dithiobis-2-nitrobenzoic acid

7. Plant material & ekstrak preparation
Piper betle
Plant material & ekstrak preparation
Preparation of ethanolic extract of Piper betle
Dosage fixation
Determination of total phenolic & flavonoid contents

8. Animals & experimental design
Male albino Wistar rats
Body weight g
The rats were allowed standar pellet diet and water adlibitum for the duration of the experiment
Animals & experimental design
24 ekor
6 ekor G1 G2 G3
PBE
(200 mg/kg/day)
Normal saline G4 Cd
(5 mg/kg/day)
Cd (5 mg/kg/day) +
PBE (200 mg/kg/day)

9. Activities of serum marker enzymes
Biochemical assays
Activities of serum marker enzymes
Determination of lipid peroxidation & oxidative stress markers
Determination of non-enzymatic & enzymatic antioxidants
Estimation of membrane-bound ATPase

10. Histopathological studies
Tissue samples
Sections of tissues
(5-6 µm thick)
Fixed in 10 % formalin-saline
Prepared by using a rotary microtone
Dehydrated by passing (mixture of ethyl alcohol, water)
Stained with hematoxylin & eosin dye
Cleaned in xylene
Which was mounted in a neutral deparaffine xylene medium for microscopical observations
Embedded in paraffin

11. Statistical analyses
Data were analyzed by one way analysis of variance (ANOVA) followed by Duncan’s mutiple range test (DMRT) using commercially available statistics software package (SPSS for Windows, V.13,0 Chicago, USA). Result were presented as mean ± SD. p value < 0,05 were regarded as statistically significant.

12. Result and discussion

13. Effects on body weight gain, relative liver weight,
feed and water intake

14. Effects on serum hepatic marker enzymes and bilirubin
Lanjutan

16. Effects on hepatic oxidative stress markersNs

17. Effects on hepatic non-enzymatic antioxidantsNs

18. Effects on hepatic enzymatic antioxidantsNs

19. Effects on hepatic membrane bound ATPasesNs

20. Histological examination
of liver tissue
A : Control
B : Cd treated rats
C : Cd treated rats
D : Cd+PBE treated rats
E : PBE treated rats

21. Conclusion
Cd is capable of causing marked oxidative stress in addition to deplete the antioxidants and inhibiting the activities of antioxidants enzymes.
The treatment with PBE could significantly attenuate the Cd induced oxidative hepatotoxicity and shows its therapeutic potential to be used as a cost effective safe herbal oxidative agent in the treatment of Cd toxicity.