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CARDIOPROTECTIVE ROLE OF TERMINALIA CATAPPA.Linn AND CISSAMPELOS PAREIRA. Linn ON DOXORUBICIN        INDUCED CARDIAC TOXOC...
 AIM AND OBJECTIVE OF THE STUDY The Goal of the present study is to investigate• Cardiac protective activity of Terminal...
PLAN OF WORK    Plant selection and acquisition        Botanical identification       Drying of plant materials     Extrac...
INTRODUCTION• Cardiovascular diseases remain the principal cause of death in  both developed and the developing countries....
• Traditionally, Terminalia catappa.Linn and Cissampelos  pareira.Linn were used as cardio-tonics and pharmacologically  b...
MYOCARDIAL INFARCTION• Myocardial infarction is an ischemic necrosis of a portion of the   myocardium due to sudden occlus...
 EPIDEMOLOGY:• MI is a common occurrence in young man(40-60yrs) and in  women (60-85yrs).• World wide more than three mil...
 AEITOLOGY:               SYMPTOMS:• Hypercholesterolemia                          • Pain• Diabetes mellitus            ...
 Pathophysiology:                     Role of plaque in MI
 DIAGNOSIS OF MYOCARDIAL INFARCTION• The diagnosis of myocardial infarction can be made after  assessing patients complai...
 MYOCARDIAL INFARCTION COMPLICATIONS• Arrhythmias• Heart failure• Pericarditis• Emboli
PLANT PROFILETerminalia catappa.L   Cissampelos pareira.L
 TAXONOMY:                  TAXONOMY:Kingdom    :Plantae         Kingdom    : PlantaeDivision   :Magnoliophyta   Divisio...
UsesTerminlia catappa.Linn                    Cissampelos pareira. Linn It has been used for dysentry,           It has ...
REVIEW OF LITERATURE                             Terminlia catappa. lS.NO          Authors & year                         ...
Cissampelos pareira.LS.NO          Authors & year                       Activity done1      Singh BK et al., (2012)       ...
• MATERIALS:Animals: Wistar rats(150-200gms)         Swiss albino mice(20-30gms)  S.NO     Chemical                       ...
• METHODS Identification and authentification of plants Collection of plant materials preparation of plant materials fo...
ACUTE TOXICITY STUDIES The procedure was followed by using OECD 423 (Acute Toxic   Class Method). The starting dose leve...
PHARMACOLOGICALSCREENING GROUPING:• Group I : Control (Normal control)• Group II : Positive control(doxorubicin(15 mg/kg ...
• Group VII: Test V(DOX+Plant Extract-250mg/kg B.W of  EETC)• Group VIII: Test VI(DOX+Plant Extract-500mg/kg B.W of  EETC)...
• Rats were divided into ten groups each group contain six animals.• Group I serves as normal which receives normal saline...
• Group VII, Group VIII, Group IX and Group X also serves as test  and receives doxorubicin(15 mg/kg B.W) first and then r...
RESULTS Phytochemical analysis:   S.NO     COMPOUNDS            ETHANOLIC     EXTRACT       OF ETAHANOLIC       EXTRACT  ...
 Acute toxicological study:• From acute toxicity studies their was no mortality at a dose of  5000mg/kg B.W.
 Pharmacological screening:  Effect of ethanolic extracts of Terminalia catappa. L and  Cissampelos pareira. L on serum b...
Effect of ethanolic extracts of EETC and EECP on antioxidant levels in doxorubicin treated rats hearts    S.NO    GROUPS  ...
Effect of Terminalia catappa.L and Cissampelos pareira.L on serum CK, LDH, SGOT & SGPT levels in doxorubicin induced cardi...
Histopathlogical studies
CONCLUSION• The present study suggest that the         Terminalia catappa.L and  Cissampelos pareira.L could be used as an...
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  1. 1. CARDIOPROTECTIVE ROLE OF TERMINALIA CATAPPA.Linn AND CISSAMPELOS PAREIRA. Linn ON DOXORUBICIN INDUCED CARDIAC TOXOCTIY IN RATS THESIS Submitted to Jawaharlal Nehru Technological University, Anantapur, In the partial fulfillment of the requirements for the award of the degree of MASTER OF PHARMACY IN PHARMACOLOGY By Y.LALITHA [Reg.No.10AF1SO108] Under the guidance of V. JAYA SANKAR REDDY M.Pharm, (Ph.D) Department of PharmacologyKRISHNATEJA PHARMACY COLLEGE, TIRUPATHI, CHITTOOR (Dt) SEPTEMBER -2012
  2. 2.  AIM AND OBJECTIVE OF THE STUDY The Goal of the present study is to investigate• Cardiac protective activity of Terminalia catappa.L and Cissampelos pareira.L against doxorubicin induced cardiac toxicity in rats. The following objectives were set to achieve the goal of the present study• To perform the acute toxicity studies of ethanolic extracts of Terminalia catappa.L and Cissampelos pareira.L on mice.• To investigate the effect of various doses of Terminalia catappa.L and Cissampelos pareira.L on serum biomarkers (CK, LDH, SGOT, SGPT) in normal and cardio toxic rats.• To investigate the effect of various doses of Terminalia catappa.L and Cissampelos pareira.L on antioxidant enzymes (SOD and catalase) in normal and cardio toxic rats.
  3. 3. PLAN OF WORK Plant selection and acquisition Botanical identification Drying of plant materials Extraction of plant materials Phytochemical profiling Acute toxicological studiesScreening of cardio protective activity
  4. 4. INTRODUCTION• Cardiovascular diseases remain the principal cause of death in both developed and the developing countries.• Myocardial infarction is a prominent implication of the cardiovascular disease. According to WHO 16.7 million people die each year owing to heart attacks.
  5. 5. • Traditionally, Terminalia catappa.Linn and Cissampelos pareira.Linn were used as cardio-tonics and pharmacologically both shown significant antioxidant property, hence these plants were chosen to evaluate the Cardio protective Activity.• Doxorubicin, a successful antitumor drug also known for its cumulative and dose dependent cardio toxicity. Hence Dox has been chosen for the study to induce cardio toxicity.
  6. 6. MYOCARDIAL INFARCTION• Myocardial infarction is an ischemic necrosis of a portion of the myocardium due to sudden occlusion of a branch of coronary artery. CLASSIFICATION: . ST elevation MI(STEMI) . Non- ST elevation (non-STEMI)
  7. 7.  EPIDEMOLOGY:• MI is a common occurrence in young man(40-60yrs) and in women (60-85yrs).• World wide more than three million people were suffering with STEMI and more than four million people were suffering with Non-STEMI.• Blacks and whites are equally affected.• Mortality estimates due to CVD vary widely by state, ranging from 10% in Meghalaya to 49% in Punjab.
  8. 8.  AEITOLOGY:  SYMPTOMS:• Hypercholesterolemia • Pain• Diabetes mellitus • Difficulty in breathing• Smoking • Dizziness• Stress• Obesity • Nausea• Insufficient exercise • Shortness of breath• Hypertension • Mimics like MI
  9. 9.  Pathophysiology: Role of plaque in MI
  10. 10.  DIAGNOSIS OF MYOCARDIAL INFARCTION• The diagnosis of myocardial infarction can be made after assessing patients complaints and physical status.• ECG changes, Coronary angiogram and levels of cardiac markers help to confirm the level of infarction.
  11. 11.  MYOCARDIAL INFARCTION COMPLICATIONS• Arrhythmias• Heart failure• Pericarditis• Emboli
  12. 12. PLANT PROFILETerminalia catappa.L Cissampelos pareira.L
  13. 13.  TAXONOMY:  TAXONOMY:Kingdom :Plantae Kingdom : PlantaeDivision :Magnoliophyta Division : MagnoliophytaClass :Magnoliopsida Class : MagnoliopsidaOrder :Myrtales Family : MenispermaceaeFamily :Combrataceae Genus : CissampelosGenus : Terminalia Species : PareiraSpecies :catappa
  14. 14. UsesTerminlia catappa.Linn Cissampelos pareira. Linn It has been used for dysentry,  It has been used for heart rheumatoid arthritis, cough, problems,asthma,inflammation asthma, leprosy, nausea, eye , malaria, STD, snake bite and problems, liver problems and Kidney problems. STD.  It has been used as diuretic, It is used as cardiotonic, diuretic, antiseptic and anti periodic antipyretic, aphrodisiac and antimicrobial agent. property.
  15. 15. REVIEW OF LITERATURE Terminlia catappa. lS.NO Authors & year Activity done1 B.V.Jawal et al.,(2012) Antioxidant potential2 Abulmanzur et al.,(2011) Antimicrobial activity3 A. Aimola et al., (2011) Erythropoiesis4 Azrul et al.,(2011) Anthelmintic potential5 Paul chidokachikezie (2011) Antipolymerization6 Saheb L and SS Wadje.,(2011) Fungicide activity7 Saroja M et al.,(2011) Antioxidant activity of phenolic fraction8 H.V Annegowda et al.,(2010) Analgesic and antioxidant properties9 Jing gao et al.,(2010) Hepatoprotective activity Kinoshita S et al.,(2007)10 Minakshi G Joshi et al., (2008) Antibiotic11 Ahmed SA et al.,(2006) Antidiabetic activity12 Fan YM et al., (2004) Phytochemical and anti-inflammatory activity
  16. 16. Cissampelos pareira.LS.NO Authors & year Activity done1 Singh BK et al., (2012) Cardiac protective activity2 Mahendra sigh Yadv Hepatoprotective activity etal.,(2011)3 PadminiShukla et al., (2011) In-vitro anthelmintic activity4 Pramodine d et al., (2011) Memory enhancing activity5 Surendran S et al., (2011) Hepatoprotective activity6 Gupat M et al.,(2010) Antipyretic activity7 Bafna A, Mishra S.,(2009) Immunomodulatory activity8 Amresh G et al., (2008) Toxicological screening9 Amresh G et al., (2007) Antinociceptive and antiarthritic10 G. Amresh et al., (2011) Gastroprotective11 AmreshG,Reddy GD et al., Antioxidant activity (2007)
  17. 17. • MATERIALS:Animals: Wistar rats(150-200gms) Swiss albino mice(20-30gms) S.NO Chemical Manufacture 1 Doxorubicin Celon 2 Normal saline Claris life sciences Ltd 3 CK-MB kit span diagnostics Ltd 4 SGOT kits span diagnostics Ltd 5 SGPT kits span diagnostics Ltd 6 Formalin Sd-fine chem. Limited 7 Potassium dihydrogen phosphate Merk specialities 8 Ethanol Changshuyangyuan chemicals 9 Disodium hydrogen phosphate Fisher scientific
  18. 18. • METHODS Identification and authentification of plants Collection of plant materials preparation of plant materials for the extraction Extraction procedure Preliminary phytochemical studies
  19. 19. ACUTE TOXICITY STUDIES The procedure was followed by using OECD 423 (Acute Toxic Class Method). The starting dose level of Terminalia catappa.L and Cissampelospareira.L was 2000 and 5000mg/kg body weight p.o. Dose was administered to overnight fasted mice`s. Food was withheld for and further 3-4 hours after administration. The body weight of the mice`s before and after administration were noted. the changes in the body weight were not so prominent. The onset and signs of toxicity was also not observed. No mortality was observed.
  20. 20. PHARMACOLOGICALSCREENING GROUPING:• Group I : Control (Normal control)• Group II : Positive control(doxorubicin(15 mg/kg B.W, through I.P))• Group III: Test I (Plant Extract-250mg/kg B.W of EETC+DOX)• Group IV : Test II (Plant Extract-500mg/kg B.W of EETC+DOX)• Group V : Test III(plant extract-250mg/kg B.W of EECP+DOX)• Group VI : Test IV (Plant extract-500mg/kg BW of EECP+DOX)
  21. 21. • Group VII: Test V(DOX+Plant Extract-250mg/kg B.W of EETC)• Group VIII: Test VI(DOX+Plant Extract-500mg/kg B.W of EETC)• Group IX: Test VII(DOX+ Plant extract-250mg/kg B.W of EECP)• Group X: Test VIII(DOX+ Plant extract-500mg/kg B.W of EECP)
  22. 22. • Rats were divided into ten groups each group contain six animals.• Group I serves as normal which receives normal saline(5ml/kg B.W) for 30 days and Group II serves as positive control which receives doxorubicin 7.5mg/kg B.W in two equal injections at 27thand 29th day(15mg/kg B.W; I.P route)and normal saline(1ml/kg B.W).• Group III, Group IV, Group V and Group VI serves as test which receives plant extracts of Terminalia catappa.L and Cissampelos pareira.L at dose of 250 and 500 mg/kg B.W, through oral route throughout the study period and also receives doxorubicin (15 mg /kg B.W) at 27thand 29thday.
  23. 23. • Group VII, Group VIII, Group IX and Group X also serves as test and receives doxorubicin(15 mg/kg B.W) first and then receives test extracts at an dose of 250 and 500mg/kg B.W, through oral route throughout the study period.• After the study period animals were sacrificed by collecting blood through cardiac puncture and collected blood was centrifuged and serum was collected and biochemical analysis were performed for serum levels CK-MB, LDH, SGOT, SGPT, SOD and Catalase.• The hearts were isolated and placed in 10% formalin and histopathological studies were performed.
  24. 24. RESULTS Phytochemical analysis: S.NO COMPOUNDS ETHANOLIC EXTRACT OF ETAHANOLIC EXTRACT OF TERMINALIA CATAPPA CISSEMPELOS PAREIRA 1 Carbohydrates + + 2 Fixed oils + + 3 Alkaloids + + 4 Glycosides + + 5 Steroids + + 6 Flavonoids + + 7 Tannins + + 8 Phenolic compounds + + 9 Amino acids - - 10 Saponins + + +VE: Presence of compound -VE : Absence of compound
  25. 25.  Acute toxicological study:• From acute toxicity studies their was no mortality at a dose of 5000mg/kg B.W.
  26. 26.  Pharmacological screening: Effect of ethanolic extracts of Terminalia catappa. L and Cissampelos pareira. L on serum biomarkers in normal and doxorubic in exposed rats S.NO Groups CK-MB LDH SGOT SGPT 1 Group I 126.20±2.584 116.79±2.412 68.6±4.188 46.213±0.7026 2 Group II 246.52±6.609 234.76±4.889 161.98±0.7525 92.82±1.898 3 Group III 181.00±2.369*** 143.22±4.770*** 140.23±1.074*** 81.35±0.7794*** 4 Group IV 152.80±7.061* 136.65±3.484*** 116.68±1.145*** 68.68±0.8520*** 5 Group V 171.49±2.513*** 137.22±4.019*** 130.80+4.414*** 73.67±1.433*** 6 Group VI 156.10±2.768** 120.49±5.220*** 87.53±6.7332*** 57.23±0.9472*** 7 Group VII 223.28±8.677*** 198.25±3.146*** 178.51±2.785** 78.32±0.8053*** 8 Group VIII 192.84±5.893*** 178.52±2.276*** 146.14±1.837* 69.21±1.686*** 9 Group IX 178.12±3.503*** 147.22±2.742*** 126.61±4.753*** 68.88±1.879*** 10 Group X 149.70±2.876* 133.86±2.883*** 88.248±3.664*** 51.84±1.501***
  27. 27. Effect of ethanolic extracts of EETC and EECP on antioxidant levels in doxorubicin treated rats hearts S.NO GROUPS SOD CATALASE 1 Group I 7.1±0.03890 11.24±0.1472 2 Group II 3.747±0.2449 5.07±0.1451 3 Group III 5.168±0.08392*** 7.75±0.2808*** 4 Group IV 5.597±0.2800*** 9.12±0.1183*** 5 Group V 4.980±0.2493** 8.30±0.2516*** 6 Group VI 6.112±0.1657*** 10.250±0.07638*** 7 Group VII 4.942±0.1241** 6.74±0.1700*** 8 Group VIII 4.92±0.2554** 7.64±0.1148*** 9 Group IX 4.97±0.1179** 8.54±0.2683*** 10 Group X 5.900±0.3246*** 10.81±0.2629***All values were expressed as mean+ SEM; n= 6 observation, values were significant when compared with positive control, * P<0.05,** P<0.01,*** P<0.001(one way ANOVA followed by Turkey`s test)
  28. 28. Effect of Terminalia catappa.L and Cissampelos pareira.L on serum CK, LDH, SGOT & SGPT levels in doxorubicin induced cardiac toxic rat
  29. 29. Histopathlogical studies
  30. 30. CONCLUSION• The present study suggest that the Terminalia catappa.L and Cissampelos pareira.L could be used as an antioxidant during or after doxorubicin therapy plants shows a greater protective effect in preventive as compared with curative model in rats.• Further studies should be required for the detection, isolation and structural elucidation of the chemical constituents and further investigation should be required to find exact mechanism of action for cardioprotective activity of Terminalia catappa.L and Cissampelos pareira.L plants.

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