PROPOSAL SEMINAR

PROTECTIVE EFFECT OF PROPOLIS IN KIDNEY MALE MICE

(Mus musculus) INDUCED BY LEAD ACETATE

CITRASARI HENRA PUTRI

061311133266

(Dr. Wiwik Misaco Y., drh., M.Kes) (Dr. Hani Plumeriastuti,drh., M.Kes)
Supervisor Co-Supervisor
Research
Background
Lead (Pb)
Free
Radicals

Oxidative
Stress

Antioxidant
Renal
Damage
 Problem Statement
Does the propolis extract can protect the morphological
structure of glomerulus, tubules, and papilla from lead (Pb)
acetate contamination?
Theoretical
Base
Lead ROS Renal Damage

Glomerulus Tubules Papilla

Cellular Interstitial
Acute Tubular Necrosis
Nephritis
(Nath et al, 2012)
(Missoun et al, 2010)

Propolis
Propolis
• Antioxidant activity : Phenolic acid and Flavonoid, that will neutralize and detoxify
oxidants by intracellular enzyme (Daleprane et al, 2013)
• Anti-inflammatory activity : Cinnamic acid derivatives, Flavonoid, CAPE or Caffeic
Acid Phenyl Ester will exhibit the anti-inflammatory activity (Wagh, 2013; Silva-
carvalho et al, 2015)
• Anti-bacterial activity : CAPE will block the substrate entrance of the bacteria and
prevent them them to get to the active site (Silva-carvalho et al, 2015)
• Anti-viral activity : Flavonoids are the major biactive compounds against viral
infection (Silva-carvalho, 2015)
• Anti-angiogenic activity : Polyphenols in EEP of Brazilian propolis will decrease the
production of ROS (Daleprane et al, 2013)
 Hypothesis

“Propolis extract have protective effect in renal

tissue damage that induced by lead acetate.”
 Aim of Research
To prove the protective effect of Propolis extract in protecting
the renal tissue damage of renal histopathology of mice

(Mus musculus) that induced by lead acetate.

 Outcome of Research
The result of this research will provide new information about
the benefits of Propolis extract for protecting the kidney from
cellular damage as an alternative medicine from the lead
acetate toxicity.
Material and
Methods
 Research Location and Time
• Experiment : Laboratory of Experimental Animals, Faculty of Veterinary
Medicine, Universitas Airlangga
• Extraction of Propolis : BPKI (Balai Penelitian dan Konsultasi Industri),
Ketintang, Surabaya
• Histopathological examination : Department of Pathology, Faculty of
Veterinary Medicine, Universitas Airlangga
 Research Materials
• Lead Acetate
• Ethanolic Extract of Propolis
• 1.5% of CMC Na and Tween 80 to make the solution of propolis extract
• 10% Neutral Buffered Formalin for tissue fixation
• 70, 80, 90, 96% alkohol; xylol; paraffin; entellan; and hematoxylin eosin to
make the histopathological preparat
 Research Equipment
• Treatment: Scale, Mice cage, Feed and drink container, Analytical scale,
Intubation needle and 1 ml tuberculin syringe
• Organ fixation: Surgical scissors, forceps, scalpel, gloves, plastic pots, object
glass, microscope and camera for documentation
• Histopath preparation: Microtome, water bath, hot plate, a series of
dehydrationapparatus
Preparation of Propolis Extract Solution
• EEP was dissolved in Tween 80
• Then, mixed with the mucilago of 1.5% CMC-Na
• Given in 0.01 ml/g bw of mice with dose of 200 mg/kg bw (T1),
400 mg/kg bw (T2), 800 mg/kg bw (T3)

(Proboningrat, 2015)
Preparation of Lead Acetate Solution
• Dose of Lead Acetate Solution : 20 mg/kg bw (Difitri, 2010)
• It is made from 0.02 mg dissolve in 100 ml aquadest
• It will be given one hour after propolis extract
administration, in 0.01 ml/g bw of mice
 30 ♂ Mice is Randomly Divided

C– C+ T1 T2 T3

Adaptation for 7 days

Propolis 200 Propolis 400 Propolis 800

mg/kg for 3 mg/kg for 3 mg/kg for 3
C– C+ days days days

CMC Na + CMC Na +
Tween + Tween + T1 Propolis T2 Propolis T3 Propolis
Aquadest Aquadest + Pb 200 mg/kg bw 400 mg/kg bw 800 mg/kg bw
+ Pb 20 mg/kg + Pb 20 mg/kg + Pb 20 mg/kg
bw bw bw

After 35 days, mice will be sacrified

Kidney collection

Kidney fixation

Histophatological Preparation of Kidney

Scoring of Kidney Histopathology

Data Analysis
 Scoring of Kidney Histopathology
Parameters Score

a. Granular cast
No changes 0
Mild (changes <30%) 1
Moderate (changes 30% – 50%) 2
Severe (changes >50%) 3
b. Cellular cast
No changes 0
Mild (changes <30%) 1
Moderate (changes 30% – 50%) 2
Severe (changes >50%) 3
c. Protein cast
No changes 0
Mild (changes <30%) 1
Moderate (changes 30% – 50%) 2
Severe (changes >50%) 3
 Parameters Score

d. Hydropic degeneration
No changes 0
Mild (changes <30%) 1
Moderate (changes 30% – 50%) 2
Severe (changes >50%) 3
e. Pycnotic cell
No changes 0
Mild (changes <30%) 1
Moderate (changes 30% – 50%) 2
Severe (changes >50%) 3

(Arsad et al, 2014)

Thank you for your
attention.
 Research Background
Environmental pollution Lead as The Most Common Environmental
Pollutant Caused Oxidative Stress Antioxidant Antioxidant
in Propolis Flavonoid
(Melaleuleca Journal, 2013)