DABBAGH, DAGDAGAN, DELA PENA, DIAZ, DOMINGO, DUMALEN, ENRIQUEZ, FELIPE, GINETE, GUINA, KALAW, LAGGUI

Cardiovascular

diseases (CVDs) are the leading

causes of death and disability in the world.
In 2008, an estimated 17.3 million people died from CVD, representing 30% of all global deaths.
Of these deaths, 7.3 million were due to coronary heart disease and 6.2 million were due to stroke
Source:Global Atlas on Cardiovascular Disease Prevention and Control, World Health Organization, 2011

Cardiovascular

diseases (CVDs) are the leading

causes of death and disability in the world.
Low and middle-income groups are greatly affected with over 80% of CVD deaths taking place in these groups. In 2030 CVD will remain as the single leading cause of death with almost 23.6 million people expected to die from it.
Source:Global Atlas on Cardiovascular Disease Prevention and Control, World Health Organization, 2011

In the Philippines, the Department of Health reported that heart diseases remain to be the top leading cause of mortality in the recent 5-year average.

Source: Department of Health, Philippines http://www.doh.gov.ph

Acute myocardial infarction, commonly

known as a heart attack, is the irreversible necrosis of heart muscle secondary to prolonged ischemia.
- (Zafari M, et al. 2012).

Synthetic drugs have

been effective in managing the conditions, but the side effects of the drugs do not weigh their effectiveness.
For example, Aspirin is well known for the prevention of cardiovascular thrombotic events in patients at risk for cardiovascular disease (CVD), however, it carries an increased risk for

gastrointestinal (GI) injury (e.g., ulceration).

The use of herbal medicines and research on newer ones have been encouraged in the country as part of efforts to find

cheaper yet effective

medicines, which will be affordable to the majority of the population.

flavonoids found in the Solanaceae

antioxidant, anti-allergic, anti-inflammatory, anti-microbial and anti-cancer properties.
family, are proven to have:

Eggplant (Solanum
melongena L.) is a
nutritious, widelyavailable, locally-grown, popularly consumed vegetable in the Philippines.

It is ranked as one of the top ten in terms of oxygen radical scavenging capacity due to its phenolic constituents
-(Jung et al., 2011).

Foods rich in antioxidants play an essential role in the prevention of chronic and degenerative diseases such as

cardiovascular diseases and cancer

- (Han et al., 2003).

Studies have also shown that eggplant extracts inhibit inflammation that leads to

atherosclerosis
-(Han et al., 2003).

economical alternative treatment to cardiovascular disorders waiting to be

discovered.

Given these facts and because of its abundance in the Philippines, eggplants may prove to be the safe, effective and

OBJECTIVES OF THE STUDY

To determine if flavonoids from Solanum melongena L (Eggplant) peel extract have a cardioprotective effect on male Sprague Dawley rats with Isoproterenol induced acute myocardial infarction.

1. To compare the histopathologic features of the myocardium of male Sprague-Dawley rats treated with flavonoids from Solanum melongena L. (Eggplant) peel extract from those treated with Trimetazidine after induction of myocardial infarction with Isoproterenol.

2. To compare the histopathologic features of the myocardium of male Sprague-Dawley rats treated with Trimetazidine after induction of myocardial infarction with Isoproterenol from that of the Negative Control.

3. To compare the histopathologic features of the myocardium of male Sprague-Dawley rats treated with flavonoids from Solanum melongena L. (Eggplant) peel extract from after induction of myocardial infarction with Isoproterenol from that of the Negative Control.

4. To compare the mortality rate of Sprague Dawley rats with Isoproterenol-induced myocardial infarction treated with flavonoids from Solanum melongena L. (Eggplant) peel extract to those treated with Trimetazidine.

Development of an effective, safe and affordable alternative to currently available cardioprotective drugs.

Reduction in morbidity and mortality from cardiovascular diseases.

Optimal utilization of resources.

Providing reference for future research.

METHODOLOGY

This study utilized an experimental

design to evaluate the cardioprotective

effect of flavonoids from S.melongena L (eggplant) peel extract on Isoproterenolinduced male Sprague-Dawley rats.

Forty-Five Sprague-Dawley male rats weighing 100-150 grams The test subjects were equally divided and randomly assigned via the Fishbowl method into three groups with 15 rats per group.

Collection and Verification of Plant Material Extraction of Flavonoids Testing for Cardioprotective Effect

The fruits of Solanum melongena L. or commonly known as eggplant were obtained from Cabugao, Ilocos Sur.

Add photo!

Add photo!

The fruits were examined and subjected for authentication by the Bureau of Plant and Industry.

Screening of Flavonoids from Solanum melongena L.

Bate-Smith and Metcalf Test for Leucoanthocyanins Wilstatter Cyanidin Test
Limuaco, O.M. et al, Laboratory Manual in Pharmacognosy. Manila: Centro Escolar University School of Pharmacy, 2008 24-30

Animal Treatment Groups

Fishbowl method
Group 1: Negative Control Group Group 2: Positive Control Group Group 3: Experimental Group

Administration of flavonoid extract Administered via gastric gavage for a period of 14 days. Vol of extract = Wt. of rat (kg) x dose (mg/kg) conc. of extract (mg/ml)

Rats were weighed on a weekly basis.

Administration of positive control (Trimetazidine)

10mg/kg once a day via gastric gavage for 14 days.

Induction of MI with Isoproterenol

Prior to induction, the rats were weighed individually.

85mg/kg was injected subcutaneously on the dorsal part of the body on days 14 and 15 of the study.

Cardiac Troponin T

12 hours after all rats were sacrificed via cervical decapitation Intracardiac aspiration of 2mL of blood from 4 rats from each group Qualitative evaluation of serum Troponin T levels using Roche Troponin T kit
Guidelines set forth by the Institutional Animal Care and Use Committee (IACUC) were followed.

Histopathologic features of myocardium

10% buffered formalin,

subsequently sent to UP Manila Pathology Department for histopathology tissue preparation. Five-micrometer thick sections of each rat heart were prepared and stained with Hematoxylin and Eosin.

Histopathologic features of myocardium

Slides were randomly labelled and were sent to a credible pathologist for histopathological examination

Histopathologic features of myocardium

Microscopically the lesions of the heart were graded using the following: Grade-0: No lesion; Grade-1: Fibroblastic swelling or proliferation and accumulation of histiocyte; Grade-2: Edema mottled staining, fragmentation and segmentation of muscle fiber; .

Histopathologic features of myocardium

Microscopically the lesions of the heart were graded using the following: Grade-3: Vascular and fatty degeneration, granular disintegration and hyaline necrosis of muscle fibers. Marked capillary dilatation with hemorrhage, extensive edema Grade-4: Confluent lesion throughout the heart. Lesions were similar in characteristics to those in Grade 3.

DATA ANALYSIS

Chi-square test was used to compare the mortality proportions between positive control group and the experimental group.
Treatment * Dead Cross tabulation
Count

Dead

Alive
Treatment Total Group 2 (Positive Control) Group 3 (Experimental) 11 12 23

Dead
4 3 7

Total 15 15 30

Chi-Square Tests
Value Pearson Chi-Square Continuity Correctionb Likelihood Ratio Fisher's Exact Test Linear-by-Linear Association N of Valid Cases
b. Computed only for a 2x2 table

df 1 1 1 1

Asymp. Sig. (2-sided)

.666 1.000 .666

Exact Sig. (2-sided)

Exact Sig. (1-sided)

.186a .000 .187 .180 30

1.000 .671

.500

a. 2 cells (50.0%) have expected count less than 5. The minimum expected count is 3.50.

There is no significant difference between the proportion of dead and living mice for the positive control and experimental group
(X2(df=1, n=30) = 0.186, p = 0.50 > 0.05, ns)

Chi-square test was used to compare the frequency of myocardial infarction in rats from the different groups by histopathologic results
Treatment * MI Crosstabulation Count MI

Total
.00 2.00 0 3 3 1.00 15 12 27 15 15 30

Treatment
3.00 Total

Chi-Square Tests
Value Pearson Chi-Square Continuity Correctionb Likelihood Ratio Fisher's Exact Test Linear-by-Linear Association N of Valid Cases 3.222 30 1 .073 3.333a 1.481 4.493 df 1 1 1 Asymp. Sig. (2-sided) .068 .224 .034 .224 .112 Exact Sig. (2- Exact Sig. (1sided) sided)

a. 2 cells (50.0%) have expected count less than 5. The minimum expected count is 1.50. b. Computed only for a 2x2 table

(X2(df=1, n=30) = 3.333, p = 0.112 > 0.05, ns)

One-way ANOVA , Duncans and Tukeys test histopathologic features of the three groups.
Tests of Between-Subjects Effects
Dependent Variable: Grade Source Corrected Model Intercept Treatment Error Total Corrected Total Type III Sum of Squares 14.444a 153.089 14.444 37.467 205.000 51.911 df 2 1 2 42 45 44 Mean Square 7.222 153.089 7.222 .892 F 8.096 171.612 8.096 Sig. .001 .000 .001

a. R Squared = .278 (Adjusted R Squared = .244)

F(2,42)=8.096,p=.001<.01

Grade
Treatment 1.00 3.00 TukeyHSDa,b 2.00 Sig. 1.00 3.00 Duncana,b 2.00 Sig. 15 1.000 2.4000 .339 15 15 15 1.000 1.0667 2.0667 2.4000 .602 N 15 15 Subset 1 1.0667 2.0667 2

Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square(Error) = .892. a. Uses Harmonic Mean Sample Size = 15.000. b. Alpha = 0.05.

RESULTS

CONCLUSION

There is no significant difference between the mortality rates and the histopathologic grading of myocardial infarction in the two treatment groups, therefore the cardioprotective effect of flavonoids from eggplant (Solanum melongena L.) peel extract is comparable to that of Trimetazidine.

RECOMMENDATIONS

1) To increase the sample size and perform the experiment on another animal species.
2) To isolate and use pure flavonoids of the S. melongena L. peel extract. 3) To determine the optimum dosage or concentration of the S. melongena L. peel extract by comparing different dosages.

4) To extend the length of Trimetazidine and S. melongena L. peel extract treatment. 5) To include another group to be induced with Isoproterenol but not treated with Trimetazidine or S. melongena L. flavonoid peel extract, as much as possible do the Isoproterenol induction in the same facility to reduce variables.

6) To consult at least 3 pathologists in reading the histopathologic slide specimens,and; 7) To observe the possible adverse effects of S. melongena L. peel extract.

Thank you!
DABBAGH, DAGDAGAN, DELA PENA, DIAZ, DOMINGO, DUMALEN, ENRIQUEZ, FELIPE, GINETE, GUINA, KALAW, LAGGUI