Pharmacology Proposal SECTION A SPECIAL CLASS

Toxic metal pollution of surface, groundwater and agricultural products has become a major concern in many countries, including Philippines.

Many studies have been conducted on finding the treatment for toxic metal poisoning.

• Common name : Turmeric, Indian-saffron

• Scientific name : Curcuma longa Linn.

• Plant family : Zingiberaceae

A yellow phenolic compound, curcumin is one of the three main curcuminoids extracted from the

rhizomes of turmeric plants.

Curcumin inhibits lipid peroxidation and cyclooxygenase-2

(COX-2) expression and induces glutathione S-transferase (GSH)

enzymes.

After 14 days of curcumin supplement in rodent, the total

GSH activity and malondialdehyde (MDA), which reflects

endogenous lipid peroxidation, were measured in liver. The curcumin

diet increased hepatic GSH by 16% and decreased MDA levels by 36%

when compared with controls. (Sharma, R. et. al. 2001)

Cadmium is one of the heavy metal. Toxic metal or heavy metals

are chemical elements with a specific gravity that is at least 5 times greater

than the specific gravity of water.

Transition metal

Cadmium Physical Properties

Chemical symbol Cd

Form White, Soft metal

Characteristics Malleable, Ductile and Flexible

Found In compound form with sulfide

Atomic weight 112.41

Cadmium

Human activities Environmental

Nickel-cadmium battery manufactories

Cigarette smoking

Plating, coating metal industries

Accumulated in rivers, lakes

Car exhaust

fumes

Soil, Fertilizers Earth’s crust

Sources of Cadmium

Target organs

Acute oral LD50

Route of exposure

Health Effects

Liver, kidney, lung, bone and small intestine.

Itai - Itai disease

Osteomalacia, Osteoporosis

Ingestion and inhalation

Kidney failure, other organ failure.

225 mg/ kg BW (Material Safety Data Sheet, http//www.Science Lab.com)

AbsorbedCadmium Intestinal lumen

Organs

LiverBind to Metallothionin

(MT)Cd -- MT

Kidney

Cd -- MT

Cadmium free form

MT released

Mucosal cells

Systemic circulation

MDA Level (product of lipid peroxidation)

Glutathione Level

Histopathological change in the liver cell

Lipid peroxidation has been established as a major

mechanism of cellular injury for cadmium toxicity. (Manca et al., 1991;

Yiin et al., 1999).

The mechanism involves a process whereby free radicals

"steal" electrons from the lipids in cell membranes, resulting in

cell damage. It most often affects polyunsaturated fatty acids.

Product : MDA

Glutathione (GSH), a powerful antioxidant found within every

cells, contains –SH group.

Their main function is the activity as an antioxidant

protecting the cells from damage caused by the free radical, H2O2.

GSH is produced mainly from liver.

Glutathione exists in reduced (GSH) and oxidized (GSSG)

states.

Liver cell Hepatocyte

Arranged into cord.

Lobule shape Hexagonal

At the corners between adjacent lobules are the portal triads.

Central vein – centre of each lobule, which is a branch of the hepatic vein.

Bile duct

Portal

vein

Hepatic

artery

Hepatic LobuleHepatic Lobule

PORTAL TRIAD

The liver tissue consists of epithelial hepatocytes, arranged into cords, separated by vascular

sinusoids.

SINUSOIDSINUSOID

Functionally, the liver can be divided into 3 zones,

based upon oxygen supply. Zone 1 encircles the portal tracts

where the oxygenated blood from hepatic arteries enters. Zone

3 is located around central veins, where oxygenation is poor.

Zone 2 is located in between.

1. To study acute toxicity of Cd acetate.

2. To evaluate the protective effect of curcumin on lipid peroxidation, glutathione and histopathological change in rat liver, induced by Cd acetate.

If found to be preventive, curcumin can be used as a prevention for

cadmium poisoning.

If curcumin is found to be preventive, it may be further used in

studies concerning other free radical agent or other heavy metals.

The database concerning curcumin can be used for further

studies Eg : Effect of curcumin on disease that causes free radical.

PHASE I

Protective effect of Curcumin against Cd

toxicity

1. Cd acetate solution of dose 100, 200, 300 mg/kg BW was prepared by

dissolving it in distilled water.

2. The curcumin extracted from turmeric plant was provided by the

Government Pharmaceutical Organization (GPO). The curcumin dose

250 mg/kg BW will be dissolved in glycerol.

Adult Wistar male rats 200-220 g

Acclimatization 5 days

12 hr light / dark cycle

Standard pellet diet + drinking water

32 Wistar Rats

10% formalin buffer

Treated (Oral administration)

Sacrificed by anesthesia

1.00 gm of Liver Remainder of liver

Frozen in -70 .c

5 days

Liver Homogenate (1.5 ml)

Add TCA- TBA- HCl solution (3 mL)

Centrifuge 1,000 rpm 10 min

Pipette supernatant

Spectrophotometer (Absorbance at 535 nm)

Heat in boiling water 15 minutes

Liver Homogenate (0.5 mL)

Add 4% Sulfosalicylic acid (0.5 mL)

Supernatant (0.5 mL)

Add 0.1 mM dithiobenzoic acid (4.5 mL)

Spectrophotometer (Absorbance at 412 nm)

Centrifugue at 1,000 rpm 10 min

FIXATION

DEHYDRATION

INFILTRATION

EMBEDDING

SECTIONING BY MICROTOME

STAININGH & E

MOUNTING SLIDE

MICROSCOPIC EXAMINATION