“Cardioprotective effect of Rhododendron arboreum on isoproterenol induced myocardial infarction”
M. Pharm Dissertation Protocol Submitted to
RajivGandhiUniversity of Health Sciences, Karnataka
Bangalore– 560 041
By
Mr. SANDIP HARESHBHAI KARIA
B.Pharm
Under the Guidance of
Dr. Divakar Goli M.Pharm. Ph.D
Professor
P.G. Department of Pharmacology,
Acharya & B.M.ReddyCollege of Pharmacy,
Soladevanahalli, Chikkabanavara (Post)
Hesaraghatta Main Road, Bangalore – 560 090
RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BANGALORE
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
1. / Name of the candidate &Address. / Mr. SANDIP HARESHBHAI KARIA
#102, NAVJIVAN APPARTMENTS,
AMBAJI MANDIR ROAD,
VERAVAL - 362266
GUJARAT
2. / Name of the Institution. / Acharya & B.M. Reddy College Of Pharmacy
Soladevanahalli, Hesaraghatta Road,
Chikkabanavara Post,
Bangalore-560090.
Phone No: 080 65650815
Fax No: 080 28393541
3. / Course of the study
& subject. / M.Pharm (Pharmacology)
4. / Date of admission. / June-2007
5. / Title of the Topic / “Cardioprotective effect of Rhododendron arboreum on isoproterenol induced myocardial infarction”
6. / Brief resume of intended work
6.1 Need of the work
6.2 Review of Literature
6.3 Aim and Objective of the study / Enclosure I
Enclosure II
Enclosure III
7. / Materials & Methods
7.1 Source of data
7.2 Methods of collection of data
7.3 Does the study require investigation on animals?
If yes give details
7.4 Has ethical clearance been obtained from your institution in case of 7.3 / Enclosure IV
Enclosure V
Enclosure VI
Yes (Copy Enclosed)
8. / List of references (About 4 – 6) / Enclosure VII
9. / Signature of the candidate
10. / Remarks of the guide
11. / Name & Designation of
11.1 Guide
11.2 Signature of Guide
11.3 Co – Guide
11.4 Signature of Co Guide
11.5 Head of the Department
11.6 Signature of HOD / Dr. Divakar Goli M.Pharm. Ph.D
Professor
P.G. Department of Pharmacology,
Acharya & B.M.ReddyCollege of Pharmacy,
Soladevanahalli, Chikkabanavara (Post)
Hesaraghatta Main Road , Bangalore – 560 090
Mr. Manjunatha. P. M. M.Pharm., (Ph. D)
Sr. Lecturer
P.G. Department of Pharmacology,
Acharya & B.M.ReddyCollege of Pharmacy,
Soladevanahalli, Chikkabanavara (Post)
Hesaraghatta Main Road , Bangalore– 560 090
Mrs.Kalyani Divakar M.Pharm. (Ph.D)
Asst. Professor
P.G. Department of Pharmacology,
Acharya & B.M.ReddyCollege of Pharmacy,
Soladevanahalli, Chikkabanavara (Post)
Hesaraghatta Main RoadBangalore– 560 090
12 / Remarks of the Principal
Signature / Principal
Acharya & B.M.ReddyCollege of Pharmacy,
Soladevanahalli, Chikkabanavara (Post)
Hesaraghatta Main Road , Bangalore– 560 090.
Enclosure - I
6. BRIEF RESUME OF INTENDED WORK
6.1 NEED OF THE WORK:
Cardiovascular disease (CVD) remains the principle cause of death in both developed and developing countries, accounting for roughly 20% of all worldwide deaths per year[1]. Myocardial infarction (MI) is the acute condition of necrosis of the myocardium that occurs as a result of imbalance between coronary blood supply and myocardial demand [2]. Myocardial ischemia – reperfusion is clinically relevant to situations such as myocardial infarction, coronary angioplasty, thrombolytic therapy, coronary revascularization and heart transplantation. The reperfusion period, although clearly beneficial for heart, is associated with myocardial injury [3]. There is substantial evidence that ischemic tissue generates oxygen-derived free radicals (oxygen radicals), i.e., oxygen molecules containing an odd number of electrons, making them chemically reactive and often leading to chain reactions [4,5]. These oxygen free radicals may results in depression in contractile function, arrhythmias, depletion of endogenous antioxidant network, membrane permeability changes resulting in an increase in myocardial malondialdehyde (MDA) content [6].
Free radicals and reactive oxygen species have been implicated in cardiac disease and metabolic disorders, which result due to exposure to chemicals and environmental agents. Isoproterenol (ISPH), a synthetic catecholamine and β-adrenergic agonist, has been found to cause a sever stress in the myocardium resulting in infract like necrosis of the heart muscle and is well known to generate free radicals and stimulate lipid peroxidation, which may be causative factor for irreversible damage to the myocardial membrane in experimental myocardial infarction [7]. Millions of adults are taking β-adrenoceptor blocker drugs to lower blood pressure, lower cholesterol, and/or to reduce platelet aggregation and the prescribed regimen must be adjusted for individual needs by modulating the drug dosage and selecting from a collection of possible drugs to yield the desired response while keeping serious side effects to a minimum.
Recently, attention has been focused on non-nutrient phytochemicals and polyphenols such as the flavonoids, alkaloids and xanthones such as the derived from different plants species a potential therapeutic agents in the prevention and management of cardiovascular diseases due to there antioxidant nature [8].
Rhododendron arboreum is an important medicinal plant, which is used for treatment of various ailments in Ayurvedic system of medicine. Flavonoids, isolated from the leaves of Rhododendron arboreum were found to have potent antioxidant property [9] and the plant Rhododendron arboreum have been reported for anti-inflammatory [10]
In the absence of reliable cardio protective drugs in modern medicine, there are numbers of medicinal preparations in the ayurvedic system of Indian medicine recommended for the treatment of cardiac disorders. Their usage is in vogue since centuries and are quite often claimed to offer significant relief. However, no scientific information is available regarding the cardioprotective effect of Rhododendron arboreum.Since, antioxidants are known to reduce the development of chemically induced myocardial infarction, the effect of ethanol extract of leaves of Rhododendron arboreum has been evaluated for cardioprotective activity.
Enclosure – II
6.2 REVIEW OF LITERATURE:
The cardioprotective effect of the ethanol extracts of Picrorrhiza kurroa rhizomes and roots (PK) on isoproterenol- induced myocardial infarction in rats with PK (80 mg kg-1 day-1 for 15 days) significantly prevented the isoproterenol-induced myocardial infraction and maintained the rats at near normal status [11].
The study was conducted to elucidate the antioxidant role of garlic oil in isoproterenol (IPL)-induced myocardial infraction in rats. In myocardial necrosis induced by isoproterenol, a significant increase in serum iron content with a significant decrease in plasma iron binding capacity, ceruloplasmin activity and glutathione (GSH) level were observed. There was also a significant increase in lipid peroxidase (GPX), glutathione-S transferase (GST) and glutathione reductase (GRD) were decreased significantly in heart with isoproterenol-induced myocardial necrosis. Garlic oil produced a marked reversal of these metabolic changes related to myocardial infarction induced by isoproterenol. In conclusion, garlic oil exerts its effect by modulating lipid peroxidation and enhances antioxidant and detoxifying enzyme system [12].
Isoproterenol induced myocardial infarction was confirmed by disturbances in serum and heart tissue marker enzymes such as lactate dehydrogenase (LDH), creatine phosphor kinase (CPK), aspertate transaminase (AST) and alanine transaminase (ALT), increased level of lipid peroxidation and Histopathological changes in the heart of isoproterenol administered rats. Pretreatment with magniferin (10 mg/100 g body weight) for 28 days was found to ameliorate the effect of isoproterenol include pathological changes, reduced the lipid peroxide formation and retained the myocardial marker enzymes activities at near normal levels. The above results indicated the cardio protective effect of mangiferin against isoproterenol induced myocardial infarction in rats [13].
Cardio protective effect of Ethanolic extract of Terminalia chebula fruits (500 mg/kg body weight) was examined in isoproterenol (200 mg/kg body weight) induced myocardial damages in rats. In isoproterenol administered rats, the level of lipid peroxides increased significantally in the serum and heart. A significant decrease was observed in the activity of the myocardial marker enzymes with concomitant increase in their activity in serum. Histopathological examination was carried out to confirm the myocardial necrosis. T. chebula extract pretreatment was found to ameliorate the effect of isoproterenol on lipid peroxide formation and retained the activities of the diagnostic marker enzymes [14].
The protective effect of rutin on isoproterenol- induced myocardial infarction in rats with PK (40mg 80 mg kg-1 day-1 for 42 days) significantly prevented the isoproterenol-induced myocardial infraction [15].
Enclosure – III
6.3 AIM AND OBJECTIVE OF THE STUDY
AIM:
In the present study, different doses of Rhododendron arboreumto be tried to assess their cardio protective effect in chemically induced myocardial infarction in albino wistar rats.
OBJECTIVE:
Evaluate the efficiency of Rhododendron arboreumin myocardial infarction injury in comparison with rutin.
Enclosure – IV
7. Materials and Methods
7.1 SOURCE OF DATA:
The source of data are from experiments on animals which involves the following :
1. Cardio protective potential of Rhododendron arboreum(sample collected by extraction of plant Rhododendron arboretum)
2. Evaluation of cardio protective activity of Rhododendron arboreumon experimental animals (male albino wistar rat).
Enclosure – V
7.2 METHOD OF COLLECTION OF DATA
- plant:Rhododendron arboreum
- Pharmacological studies:
Evaluation of cardio protective activity of Rhododendron arboreumon isoproterenol induced myocardial infarction in animals (male albino wistar rat).
- Inclusion criteria: only healthy male Wistar rats, weighing between 150-200 g should be selected.
- Rats were randomly divided into eleven groups – each consisting of EIGHT animals.
Group 1: Sham group-Control group animals given saline (1ml/rat).
Group 2: Control group animals given vehicle (1ml mg kg-1 day-1 /rat).
Group 3: Rats subcutaneously injected with isoproterenol (150 mg kg-1
day-1 /rat dissolved in saline) once a day for 2 days.
Group 4: (60 mg kg-1 day-1 /rat) Rhododendron arboreum given orally
using an intragastric tube daily for 42 days.
Group 5: (100 mg kg-1 day-1 /rat Rhododendron arboreumgiven orally
using an intragastric tube daily for 42 days.
Group 6: (60 mg kg-1 day-1 /rat) Rutin given orally using an intragastric
tube daily for 42 days.
Group 7: (100 mg kg-1day-1 /rat) Rutin given orally using an
intragastric tube daily for 42 days.
Group 8: Rats pretreated with (60 mg kg-1day-1 /rat) Rhododendron
arboreum given orally using an intragastric tube daily for 42
days and then subcutaneously injected with isoproterenol 150
mg kg-1 day-1 /rat once a day for 2 days.
Group 9: Rats pretreated with (100 mg kg-1day-1 /rat) Rhododendron
arboreumgiven orally using an intragastric tube daily for 42
days and then subcutaneously injected with isoproterenol
150 mg kg-1 day-1 /rat once a day for 2 days.
Group 10: Rats pretreated with (60 mg kg-1day-1 /rat) Rutin given
orally using an intragastric tube daily for 42 days and then
subcutaneously injected with isoproterenol 150 mg kg-1
day-1 /rat once a day for 2 days.
Group 11: Rats pretreated with (100 mg kg-1day-1 /rat) Rutin given
orally using an intragastric tube daily for 42 days and then
subcutaneously injected with isoproterenol 150 mg kg-1
day-1 /rat once a day for 2 days.
Note- 88 Animal required for carrying out cardioprotective activity and after experiment the rats will be sacrificed for collecting blood and heart tissue to carry out Bio-chemical estimations.
- Parameters are selected to study cardio protective activity in tissue and serum.
- Assay of creatinine kinase.
- Assay of lactate dehydrogenase.
- Assay of aspartate transaminase.
- Assay of alanine transaminase.
- Estimation of plasma TBARS.
- Estimation of TBARS in heart.
- Estimation of lipid hydroperoxides.
- Assay of superoxide dismutase (SOD).
- Assay of Catalase
- Assay of glutathione peroxidase
- Estimation of reduced glutathione
- Estimation of vitamin C.
- Estimation of total protein.
The data obtained from the above study will be subjected to statistical analysis using ‘t’ test.
Total duration for the completion of whole project may be 9 months
- Duration of experimentation Six months.
- Literature survey one & half months.
- Thesis writing one & half months.
ENCLOSURE – VI
7.3 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly.
The above study requires investigation on Albino Wistar Rats (male) for cardio protective activities.
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
The study is cleared from Ethical Committee of the institution. (Certificate enclosed).
ENCLOSURE – VII
8. LIST OF REFERENCES:
1. De Bono, D. P., Boon, N. A. Diseases of cardiovascular system. In: Edwards, C. R. W., Boucheir, I. A. S. (eds) Davidson’s principles of prctice and medicine. Churchill Livingstone, Hong Kong, (1992), p249-340.
2. Boudin S, Laclau M N, Tariosse, et al., Alteration of mitochondrial function in a model of chronic ischemia in vivo in rat heart, Am J Physiol Heart Circ Physiol, 282(2002)H821.
3.Ferrari R, Curello S, et al., Occurrence of oxidativestress during reperfusion of the human heart, Circulation,81(1990)201-211.
4. Muruganandan S, Gupta S, Kataria M, et al., Mangiferin protects the streptozotocin-induced oxidative damage to cardiac and renal tissue in rats, Toxicology, 176(2002)165.
5. De-Jian J, Gui-Shan T, Feng Y, et al., Protective effect of xanthones against myocardial ischemia-reperfusion injury in rats, Acta Pharmacol Sin, 24(2003)175.
6. Curello S, Cecoin C, et al ., Oxidative stress during myocardial ischemia and reperfusion: experimental and clinical evidences, Journal ofApplied Cardiology, 1(1986)311-327.
7. Senthil kumar H, Anandan R, Devaki T and Santhosh Kumar M, Cardioprotective effect of Picrorrhiza Kurrora against isoproterenol induced myocardial stress in rats, Fitoterapia , 72 (2001) 402.
8. Pauletti PM, Castor-Gamboa I, siqueira Silva DH, Young MC, Eberlin MN, New antioxidant C-glucosylxanthones from the stems of Arrabidaea samydoides, J Nat Prod, 10(2003) 1284.
9. Dhan P, Garima U, Singh BN, Ruchi D, Sandeep K, Singh KK. Free radical scavenging activities of Himalayan rhododendrons. Curr Sci 2007; 92(4): 526-32
10. Shyam S A, Kalpana S. Anti-inflammatory activity of flowers of Rhododendron arboreum (SMITH) in rat’s hind paw oedema induced by various phlogistic agents. Indian J Pharmacol 1988; 20(2): 86-9.
11.Subramanaiam Hari Seenthil Kumar, et al., Cardioprotective effects of Picorrhiza Kurroa against isoproterenol-induced myocardial stress in rats, Fitotherapia72(2001)402-405.
12. Saravanan G, Prakash J, Effect of garlic (Allium sativum) on lipid peroxidation in experimental myocardial infarction in rats, Journal of Ethnopharmacology, 94(2004)155-158.
13.Prabhu S, Mallika Jainu, et al., Cardioprotective effect of mangiferin on isoproterenol induced myocardial infarction in rats, Indian Journal of Experimental Biology, 44(2006)209-215.
14. Suchalatha S, Shyamala Devi, Protective effect of Terminalia chebula against experimental myocardial injury induced by isoproterenol, Indian Journal of Experimental Biology,2(2004)174-178.
15.M.Karthik , P.Stanely Mainzen Prince Protective role of rutin , a bioflavonoid, on lipid peroxides and antioxidants in isoproterenol-induced myocardial infarction in rats,journal of pharmacy and pharmacology,2006,701-707.