Effects Of Aqueous Extract Of Punica Granatum Seed On Triton-X100 Induced Hypercholesterolemia In Rats

Project and Seminar Material for Microbiology

Effects Of Aqueous Extract Of Punica Granatum Seed On Triton-X100 Induced Hypercholesterolemia In Rats


Abstract


The present study was carried out to investigate the effects of oral administration of aqueous extract of Punicagranatum seed on Triton X-100 induced hypercholesterolemia in rats. Hypercholesterolemia was induced by single intraperitoneal injection of Triton X-100 and the rats were later treated with aqueous extract of Punicagranatum seed at different dosage levels (50,100 and 200mg/kg body weight) and 10ml/kg/day of standard drug (Astrovastatin) for seven days. Another group received 0.9% Normal saline (1 ml/kg/day). Animals were euthanize by anaesthetization at the end of the experiment to harvest serum used for the biochemical analysis. Aqueous extract of Punicagranatum seed extract prevented triton- induced increase in serum cholesterol, triglyceride (TG), low density lipoprotein (LDL) and decrease in high density lipoprotein (HDL) in a statistically significant manner (p < 0.05) and reversed the elevated serum LDL/HDL ratio comparably to Astrovastatin, The results demonstrated the amelioration of triton- induced hypercholesterolemia in rats by aqueous extract of Punicagranatum seed. Consequently, we can hypothesize that the part of Punicagranatum seed mediated therapeutic effects is associated with its antiatherogenic/hypolidemic component.


Table of Contents


  • Title page
  • Approval page
  • Dedication
  • Acknowledgement
  • Table of Content
  • List of Table
  • List of Figure
  • Abstract

Chapter One

Introduction

  • 1.1 Background of the study
  • 1.2 Justification of the study
  • 1.3 Aim and Objectives

Chapter Two

Literature Review

  • 2.1 Punicagranatum fruits
  • 2.1.2. Botanical description of Punicagranatum seed
  • 2.1.3. Health benefits of Punicagranatum seed
  • 2.2 Cholesterol
  • 2.2.1. Hypercholesterolemia
  • 2.2.2. Description of Hypercholesterolemia
  • 2.2.3. Signs and symptons of Hypercholesterolemia
  • 2.2.4. Causes of Hypercholesterolemia
  • 2.2.5. Biomakers of hypercholesterolemia

Chapter Three

Materials and Methods

  • 3.1. Materials
  • 3.1.1. Punicagranatum fruits
  • 3.1.2. Experimental animals
  • 3.1.3. Reagent and assay kits
  • 3.2. Methods
  • 3.2.1 Preparation of aqueous extract of Punicagranatum seed
  • 3.2.2. Phytochemical analysis
  • 3.2.3. Induction of Hypercholesterolemia
  • 3.2.4. Experimental Design
  • 3.2.5. Preparation of Serum
  • 3.3. Biochemical assay
  • 3.3.1. Determination of Serum Total cholesterol
  • 3.3.2 Determination of Serum High Density Lipoprotein
  • 3.3.3. Determination of Serum Triglycerides
  • 3.3.4. Determination of Low Density Lipoprotein
  • 3.3.5. Determination of Very Low Density Lipoprotein
  • 3.3.6. Determination of Artherogenic Index
  • 3.3.7. Determination of Coronary Artery Risk

Chapter Four

Results

  • 4.1. Estimation of Total phenol content of aqueous extract of Punicagranatum seed.
  • 4.2. Estimation of radical scavenging 2,2diphenyl – 1- picrylhydrazyl of aqueous extract of Punicagranatum seed.
  • 4.3. Estimation of inhibitory activity using Lipase of aqueous extract of Punicagranatum seed
  • 4.4 Total Cholesterol and Triglyceride cholesterol
  • 4.5. High Density Lipoprotein and Very low density lipoprotein
  • 4.6. Low density Lipoprotein
  • 4.7. Atherogenic index and Coronary risk index

Chapter Five

  • Discussion
  • Conclusion
  • References
  • Appendix

List of Table


  • Table 1: Classification of hypercholesterolemia
  • Table 2: Serum HDL – Concentration range

List of Figure


  • Figure 1: Punicagranatum fruit (Al- Muammar 2012)
  • Figure: 2: Chemical structure of cholesterol
  • Figure 3: Hypercholesterolemia
  • Figure 4: Estimation of Total polyphenol content on aqueous extract of Punicagranatum seed
  • Figure 5: The IC50 value obtained by linear equation of Punicagranatum seed.
  • Figure 6: The IC50 value obtained by linear regression equation for Punicagranatum seed.
  • Figure 7: Effect of Punica granatum aqueous seed on serum cholesterol and triglyceride concentration in hypercholesterolemic rats.
  • Figure 8: Effect of Punica granatum aqueous seed on serum HDL and VLDL concentration in hypercholesterolemic rats.
  • Figure 9: Effect of Punicagranatumaqueous seed on serum LDL concentration in hypercholesterolemic rats.
  • Figure 10: Effect of Punica granatum aqueous seed on serum AI and CRI concentration in hypercholesterolemic rats.

Chapter One


Introduction

1.1 Background of study

Hypercholesterolemia is a lipoprotein metabolic disorder characterized by increase in plasma low density lipoprotein (LDL-c) and very low density lipoprotein (VLDL-c) cholesterols or decrease in plasma high density lipoprotein cholesterol (HDL-c). It was reported to be one of the most important risk factors in the development and progression of atherosclerosis that lead to cardiovascular diseases (Rerkasem et al., 2008). Hypercholesterolemia poses a major problem to many societies as well as health professionals because of the close correlation between cardiovascular diseases and lipid abnormalities (Matos et al., 2005; Ramachandran et al., 2003).

Hypercholesterolemia is highly prevalent in all the geopolitical zones of Nigeria. It ranges from 60% among apparently healthy Nigerians to 89% among diabetic Nigerians (Oguejiofor et al., 2012). Clinical trials have demonstrated that intensive reduction of plasma low density lipoprotein (LDL-c) levels could reverse atherosclerosis and decrease the incidence of cardiovascular diseases (Brown and Jessup, 1999; Ichihashi et al., 1998).

Punicagranatumcommonly known as pomegranate is a member of monogeneric family, punicacae and is mainly found in Iran which is considered to be its primary centre of origin (Facciola, 1990).Punicagranatum can be divided into several anatomical compartments including seed,juice,peal,leaf,flower,bark and root each possessing interesting pharmacological and toxicological activities. Punicagranatumhas extensively been used as a traditional remedy against acidosis,microbial infection,diarrhea (Kim and Choi, 2009).Punicagranatum seeds have also been shown to contain estrogenic compounds,oestrone and oestradiol (Kim and choi, 2009).The seeds are embedded in a white, spongy astringent pulp (Stover and Mercure, 2007). Punicagranatum is known for its high antioxidant capacity, it has been used for medicinal purposes for centuries (Syed et al 2007). Punica granatum products have also been reported to possess among others anti-inflammatory,antimicrobial and protective effects on liver function and lipid glucose metabolism(Medjakovic and jungbauer, 2013).According to Al-Muammar and khan 2015, the routine supplementation of Punica granatum may prevent or even correct obesity, diabetes and cardiovascular diseases.


1.2 Justification of study

Hypercholesterolemia is a problem faced by manysocieties since it constitutes one of the major risk factors for the development of cardiovascular diseases, such as atherosclerosis and its complications, acute infraction of themyocardium or hypertension (Robbins, 1991; Gerhardt and Gallo 1998; Gomeset al., 1998). In view of the adverse effects associated with hypercholesterolemia, the quest for natural products with lipid-lowering potential and with minimal or no side effect is needed.


1.3 Aim and Objectives of the study

The main aim of this research is to evaluate the effect of aqueous extract of Punica granatum seed on triton X-100 induced hypercholesterolemia in male albino rats.

In order to achieve the aim, the following specific objectives are followed.

  1. To estimate total phenol content in aqueous extract of Punicagranatum seed.
  2. To estimate invitro radical scavenging of power aqueous extract of Punicagranatum seed using 2, 2, diphenyl-1-picrylhydrazyl (DPPH) as substrate.
  3. To estimate inhibitory power of aqueous extract of Punicagranatum seed on lipase enzyme activity.
  4. To induce hypercholesterolemia in rats using Triton X-100.
  5. To study the effect of aqueous extract of Punica granatum seed on serum lipid profile of hypercholesterolemic rats.

Chapter Five


Discussion

Results of the present study revealed that feeding aqueous extract of Punica granatum seed resulted in significant increase in serum levels of TC, TG, LDL-c and VLDL-c accompanied with a significant decrease in HDL-c level as compared to the normal rats. The increase in serum concentrations of the above mentioned parameters and the reduction in serum HDL-c as a result of induction of hypercholesteremia with Tritonx-100 have been pointed out as risk factors for the development of atherosclerosis and related cardiovascular diseases, which was represented by the decrease in HDL/TC .

Regarding the serum LDL-c results obtained it indicated that hypercholesterolemic rats had a significant increase in serum LDL when compared with the negative control, it also shows that the oral administration of 50mg/kg,100mg/kg and 200mg/kg body weight of aqueous extact of Punicagranatumseed extact significantly decreased serum LDL-c level when compared with hypercholesterolemic rats.

Punica granatummay have exhibited cardiovascular protective activity due to its antioxidant property, Hence increasing the intake of this fruit in the diet would have an economical effect on hypercholesterolemia . Therefore Punica granatum elicited health benefits through the modulation of physiological functions including the atherogenic lipid profile.


Conclusion

Based on the findings on the results it can be concluded that the aqueous extract of Punica granatum seed ameliorate the Tritonx-100 induced hypercholesteremia in rats. Punica granatum seedaqueousextract may be beneficial forpatients who suffer from hypercholesterolemia, arteriosclelorosis.


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