Submitted To: Dr. Sammia ShahidSubmitted By: Shakeel Ahmad Khan (14003140007)

Programme: MS (Chemistry)Department of Chemistry, University of Management and

Technology Lahore

Hepatoprotective effect of Rhodiola imbricata rhizome against paracetamol-induced liver toxicity in rats

LIST OF CONTENTSIntroduction about LiverFunctions of LiverSymptoms of LiverThings Liver don’t likeDrugs that responsible for HeptotoxicityHow Paracetamol Effect LiverWorld diversion to natural productsIntroduction of Rhodiala imbricata (rhizome)Objective of this studyMaterial and Method Hepatoprotective study of Rhodiala imbricata (rhizome)Result and DiscussionConclusion, Future Aspects and References

Your Liver? Largest internal organ of the body.

The liver is a reddish brown triangular organ with four lobes of unequal size and shape.

Present at right upper side of body behind the ribs.

Weighs about 3-5 pounds

Has over 500 jobs

SOME OF MY IMPORTANT JOBS

Make you feel good Help you digest food Give you energy Clean your blood (like a filter) Help build muscles Metabolism (Synthesis and Breakdown) Excretion of waste products from bloodstream

into bile. Vascular – storage of blood

Symptoms

When your liver is sick there can be symptoms like

FatigueNauseaLoss of appetiteSwollen abdomenItchy skinJaundice

THINGS LIVER DON’T LIKE

Viruses Too many fatty foods Drugs /Medication Alcohol

DRUGS THAT INDUCED LIVER DAMAGE

Acetaminophen Carbon tetrachloride Methotrexate Anabolic Steroids Oral Contraceptive pills

HOW PARACETAMOL EFFECT LIVER It is widely used analgesic and antipyretic. But produce acute liver

damage at higher dose. Hepatotoxicity of Paracetamol has been attributed to the

formation of toxic highly reactive metabolite called n-acetylparabenzoquimine (NAPQI).

Age (years) Usual Dosage Maximum Dosage

12 years and older 325-650mg q for 4hr 4000mg/day

11-12 years 320-480mg q for 4hr 2880mg/day

6-11years 320mg q for 4hr 2600mg/day

3-6years 120-125mg q for 6hr 720mg/day

1-3years 80mg q for 4hr  

3-11 months 80mg q for 6hr  

RISK FACTOR OF LIVER AGAINST PARACETAMOL

ACETAMINOPHEN OVERDOSE METABOLISM

WHY WORLD IS DIVERTING TO NATURAL PRODUCTS

Hepatic problems are responsible for significant number of liver transplantations and deaths recorded world wide.

Pharmacotherapeutic options for liver disease are limited. Conventional or synthetic drugs used in the treatment of liver are

inadequate as well as have serious adverse effects. So it is worldwide trend to go back to traditional medicinal plants. That’s way many herbal origin products are in use for the

treatment of live ailments. Sida cordata, Solanum xanthocarpum,Phyllanthus

niruri

HEPTOPROTECTIVE EFFECT OF Rhodiala imbricata (RHIZOME)

It is a perennial herb of the family “Crassulaceae”. Commonly known as golden or arctic root. Grows on rocky slopes, common in drier areas of the western

Himalaya at an altitude of 4000-5000m. Taxonomy of Rhodiala imbricata

Kingdom Plantae

Phylum Mangoliophyta

Class Mangolliopsida

Order Saxifragales

Family Crassulaceae

Genus Rhodiola

Specie imbricata

Rhodiala imbricata

MEDICINAL IMPORTANCE Rhodiala imbricata (rhizome) was found to be have great medicinal importance due to the following Has cytoprotective efficiency.Has antioxidant ability.Wound healing ability.Has immunomodulatory ability.Has Adaptogenic activity.Has antifatigue activity.Has neuroprotective activity.Has antiproliferative activities in HT-29 colon cancer cells.Its Aqueous extract was found to be contain Gallic acid, p-tyrosol rosavin and rosin.

OBJECTIVE OF THIS STUDY Increased concerns on the side effects of current therapeutic

modalities, plants are being valued because of their curative properties and least or no side effects.

Hence an attempt has been made to assess the hepatoprotective role of Rhodiala imbricata.

MATERIAL AND METHODCollection and identification of Plant material:For this study plant material was collected from the western Himalayas, India in the month of August to September 2011.Plant material identified by and authenticated by Dr. O.P. Chaurasia, an Ethaobotanist at the Defence Institute of High Altitude Research Leh-ladkh.

MATERIAL AND METHODFollowing Chemicals (Sigma Aldrich) are

used in this study Acetone EDTA Acetaminophen Polysorbate 80 (Tween 80) Diethyl ether SilymarinFollowing Instruments are used in this study Weighing Balance Centrifuge (Japan) Auto hematology analyzer (Sysmex F-800,

Japan).

PREPARATION OF PLANT EXTRACT Freshly collected plant materials were washed under

running tap water and distilled water to remove dust particles.

Samples dried and were powdered in mechanical grinder used for solvent extraction with acetone.

CONCENTRATE OF PLANT EXTRACT

Rhodiala imbricata extract obtained after filtration was made concentrate by using Rotary Evaporator.

ANIMAL USED IN THIS STUDY Wistar albino rats (male, 150-200g) and Swiss albino mice (male

25-30g) were used.

Wistar albino rats Swiss albino mice Animals were housed in groups for 7-days for pharmacological

experiments. Animal quarter maintained at a temperature 22±2ºc and with 12h

dark and 12h light cycle. Animal had free access to commercial food and drinking water.

EXPERIMENT FOR HEPATOPROTECTIVE PROPERTY OF

Rhodiala imbricata Hepatotoxicity was induced by paracetamol induced liver damage.

Paracetamol (Acetaminophen, Sigma Chemical Company, USA) was suspended in 0.5% Tween-80 and administered p.o., at a dose of 2 g/kg.

Wistar albino rats (male) weighing between 150 and 200 g were divided into 5 groups of 6 animals each. The weight range of the animals was equally distributed throughout the groups.

Group-I served as control and received water, Group-II served as negative control, administered with paracetamol (2 g/kg, p.o.), Group-III standard, silymarin received (Sigma Chemical Company, USA) at a dose of 25 mg/kg p.o., Group-IV and V received acetone extract (200 and 400 mg/kg, p.o., respectively), once daily for 14 days.

EXPERIMENT FOR HEPATOPROTECTIVE PROPERTY OF Rhodiala imbricata

On the 14th day, blood samples were collected from all animals by puncturing retro-orbital plexus under mild ether anesthesia, later animals were sacrificed and liver tissues were collected.

The blood samples were analyzed for biochemical markers of hepatic injury and tissue samples were subjected for estimation of liver antioxidants and histopathological studies.

PREPARATION OF SERUM FROM BLOOD

Blood was drawn by puncturing the retro-orbital plexus under diethyl ether anesthesia. Whole blood for hematogram was collected in bottles containing the anticoagulant, ethylene diamine tetra-acetic acid (EDTA) while samples for biochemical analysis were collected in plain sample bottles.

Serum was separated by centrifugation at 600×g for 15 min. and analyzed for various biochemical parameters. Sera were stored in the -80°C freezer before they were analyzed.

The erythrocytes, leucocytes and platelets were determined with an Auto Hematology Analyzer (Sysmex F-800, Japan).

Alkaline phosphatase (ALP), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT) and lipid profiles like total cholesterol (TC), triglycerides (TG) and biochemical parameters such as creatinine and bilirubin were also determined using Span Diagnostics Limited kit, India.

RESULT AND DISCUSSION The effect of acetone extract of R. imbricata at two dose levels (200

mg/kg and 400 mg/kg, p.o.) on hematological parameters in paracetamol induced hepatic damage is shown in Table 1.

RESULT AND DISCUSSION Table 2 shows the effect of acetone extract (200 mg/kg and 400

mg/kg) on serum biochemical markers in paracetamol induced hepatic damage.

CONCLUSION From the present study it is evident that the acetone extract of R.

imbricata rhizome has no toxicity even at a higher dose of 2000 mg/kg.

The serum and lipid parameters were maintained at normal levels compared to control groups.

Histopathological examinations of the liver showed that extract and silymarin have a protective role over the toxicity of paracetamol.

Hence R. imbricata could be one of the best sources of natural hepatoprotective agents.

Future AspectsTo evaluation and characterization of chemical component which is

responsible for Heptoprotective activity of R. imbricata.

REFERENCES Akindele, A.J., Ezenwanebe, K.O., Anunobi, C.C., Adeyemi, O.O., 2010.Hepatoprotective and in vivo antioxidant effects of Byrsocarpus coccineusSchum. and Thonn. (Connaraceae). J. Ethnopharmacol.129, 46–52. Arora, R., Chawla, R., Sagar, R., Prasad, J., Sing, S., Kumar, R., Sharma, A., Singh, S., Sharma, R.K., 2005. Evaluation of radioproective activities of Rhodiola imbricate Edgew – a high altitude plant. Mol. Cell. Biochem. 273,209–223. Gupta, A., Kumar, R., Upadhayay, N.K., Pal, K., Kumar, R.,Sawhney, R.C.,2007. Effects of Rhodiola imbricata on dermalwound healing. Planta Med. 73,774–777. Kakkar, P., Das, B., Viswanathan, P., 1984. A modified method for assay ofsuperoxide dismutase. Indian J. Biochem. Biophy. 21, 131–132. Ramellini, G., Meldolesi, J., 1976. Liver protection by Silymarin. In vitroeffect on dissociated rat hepatocytes. Arzneim Forsch (Drug Research) 26, 69-73. Mroueh, M., Saab, Y., Rizkallah, R., 2004. Hepatoprotective activity of Centaurium erythraea on acetaminophen-induced hepatotoxicity in rats. Phytother. Res. 18, 431–433.

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hakeel Khan, Im

ran Ali (U

MT)

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