NSM 2006


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NSM 2006

  1. 1. The effect of Strobilanthes crispus extracts on atherogenesis and tissue morphology in hypercholesterolemic rabbits Nurhafzan AI 1 , Norhaizan ME 1 , Suhaila M 2 , Khozirah S 3 & Maznah I 1,4 1 Department of Nutrition & Health Science, Faculty of Medicine & Health Science, Universiti Putra Malaysia, 43400, Serdang, Selangor; 2 Department of Food Service & Management, Faculty of Food Science & Technology, UPM, 43400, Serdang, Selangor; 3 Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor; 4 Interim Institute, Pharmaceutical & Nutraceutical Biotechnology, UPM-MTDC Incubation Centre One, 43400 UPM, Serdang, Selangor
  2. 2. INTRODUCTION <ul><li>Cardiovascular disease (CVD) is the most deadly disease to the populations in the developed countries (WHO Monica Project, 1988), involving pathological changes in blood vessels associated mainly with the heart and brain. </li></ul><ul><li>Atherosclerosis is the underlying disorders in the majority of patients with CVD. </li></ul><ul><li>Atherosclerosis refers to the build up of fatty material in the arterial wall, which leads to narrowing of an artery and potential blockage. </li></ul><ul><li>This supports the occlusive blood clots formation and can give effect in injury to heart, brain and lung tissue, which may cause death (Linder, 1991). </li></ul><ul><li>Both epidemiologic and clinical trials have firmly established that hypercholesterolemia is a key risk factor in atherosclerosis while sustaining the blood cholesterol level within normal degree range can decrease the risk. (Teddy and Marcel, 2000; Jiun-Rong, 2003). </li></ul><ul><li>Antioxidants, including the natural and the synthetic antioxidants are materials which protect against free radical reaction and retard the progress of many chronic diseases such as CVD and cancer. </li></ul>
  3. 3. Strobilanthes Crispus PLANT OBJECTIVES To determine the hypocholesterolemic role of Strobilanthes crispus extracts on an animal model induced with atherosclerosis. <ul><li>Natives to countries from Madagascar to Indonesia. </li></ul><ul><li>Strobilanthes crispus plant has frequently been used as a herbal remedy locally and in Indonesia as anti-diabetic, diuretic and cholesterol lowering agents. </li></ul><ul><li>Also known as “jin batu” in Malaysia, “enyoh kelo” or “kejibeling” in Java and “daun picah beling” in Jakarta (Sunarto, 1977). </li></ul><ul><li>Water extract of Strobilanthes crispus was found to have high content of mineral, such as potassium, calcium, sodium and iron(Maznah et al ., 2000) . </li></ul><ul><li>However, currently used synthetic antioxidants have been suspected to promote negative side effects (Branen, 1975; Barlow, 1990).Therefore, people are currently interested in products contains of natural antioxidants. </li></ul><ul><li>Nowadays, many other plants species are being studied for their preventive action possibility against all kinds of diseases as well as in the search for novel antioxidants (Kim et al., 1994). </li></ul>
  4. 4. <ul><li>24 adult female New Zealand white rabbits (body weight: 1.8-2.5 kg) were used. </li></ul><ul><li>After 2 weeks of acclimatization, the rabbits were randomly assigned into 4 groups (n=6/group). </li></ul><ul><li>At the end of the experimental period (12 weeks), the rabbits were sacrificed. </li></ul><ul><li>Ascending aorta, liver, kidney and heart tissue specimens were excised immediately and prepared for the microscopic and histopathological studies by fixing in formalin and paraffin blocks. Sections were prepared and stained with haematoxylin and eosin. </li></ul>METHODOLOGY Basal diet + 0.25% HCD + Simvastatin (20 mg/kg body weight) SV Basal diet + 0.25% HCD + Strobilanthes crispus extract (SC) (0.2% of diet) SC Basal diet + 0.25% high cholesterol diet (HCD) PC Unmodified non-purified diet (Basal diet) NC Diet composition Group
  5. 5. Histology analysis of the rabbits aorta RESULT & DISCUSSION <ul><li>The intima to media thickness ratio for the aorta of the positive control group was slightly higher than the Strobilanthes crispus and simvastatin groups. </li></ul><ul><li>Although the difference was insignificant, massive macrophages and foam cells were detected in the intima lesion resulting in thickening of the positive control group aorta. </li></ul><ul><li>In contrast, the aortas from the group treated with Strobilanthes crispus extract and simvastatin showed lesser thickening than the positive control group. Both treatment groups still had some protruded intima, unlike the negative control group, as they were also induced with artherosclerosis. </li></ul>0.39  0.22 Chol + Simvastatin Values are expressed as means  sd, for n=5 0.33  0.13 Chol + S. crispus 0.67  0.57 +ve Control 0.20  0.13 Group -ve Control Intima to Media Thickness Ratio Group
  6. 6. A C D A: A photomicrograph of a section in aorta of a –ve control rabbit, H&E, 4x. B: A section in Fig A, 10x. C: A photomicrograph of a section in aorta of a +ve control rabbit, H&E, 4x. D: A section in Fig B, 10x B Tunica Adventitia TA Tunica Media TM Tunica Intima TI Lumen L
  7. 7. E H G F E: A photomicrograph of a section in aorta of a S. crispus treated rabbit , H&E, 4x. F: A section in Fig E, 10x. G: A photomicrograph of a section in aorta of a Simvastatin treated rabbit, H&E, 4x. H: A section in Fig G, 10x Tunica Adventitia TA Tunica Media TM Tunica Intima TI Lumen L
  8. 8. <ul><li>In the liver tissue, slight fatty changes and moderate to severe lymphocyte infiltration were observed in the positive control group. </li></ul><ul><li>Meanwhile, mild lymphocyte infiltration were observed in the simvastatin group. </li></ul><ul><li>Ballooning degeneration of hepatocytes in both groups were also detected. </li></ul><ul><li>Interestingly, no such changes were visible in the Strobilanthes crispus group. </li></ul><ul><li>No significant changes were observed in the heart and kidney tissues. </li></ul>RESULT & DISCUSSION CONTINUES
  9. 9. LIVER PC (20x) PC (20x) NC (20x) SC (20x) SV (20x) PC (20x)
  10. 10. HEART NC (40x) SC (40x) PC (40x) SV (40x)
  11. 11. NC (40x) KIDNEY NC (40x) SC (40x) PC (40x) SC (40x) PC (40x) SV (40x) SV (40x)
  12. 12. CONCLUSIONS REFERENCES Maznah, I., Manickam, E., Azlina, M. D., Asmah, R. and Asmah, Y. 2000. Chemical composition and antioxidant activity of Strobilanthes crispus leaf extract. J. Nutr. Biochem 11 :536-542. Sunarto, P.A. 1977. Materia Medika Indonesia. Jakarta, Indonesia: Penerbit Direktorat Jenderal Pengawasan Obat dan Makanan. The WHO Monica Project. 1988. Geographical variation in the major risk factors of coronary heart disease in men and women aged 35-64 years. World Health Stat. 41: 115-40. Linder, M. C. 1991. Nutritional biochemistry and metabolism with clinical applications (2nd ed.). New York: Elsevier. Jiun-Rong, C. and Shiau-Fang, C. 2003. Lipid metabolism in hypercholesterolemic rats affected by feeding cholesterol-free diets containing different amounts of non-dialyzed soybean protein fraction. Nutrition 19: 676-680. Teddy, T. C. and Marcel, W. L. 2000. Chinese green tea lowers cholesterol level through and increase in fecal lipid excretion. Life Science, Vol. 66. No. 5, pp. 411-423. Kim, S. Y., Kim, S. K., Oh, M. J. and Jung, M. Y. 1994. Antioxidant activity of selected oriental herb extracts. J. Am. Oil. Chem. Soc. 71: 633-640. Branen, A. L. 1975. Toxicology and biochemistry of butylated hydroxyanisole and butylated hydroxytoluene. J. Am. Oil. Chem. Soc. 52: 59-63. Barlow, S. M. 1990. Toxicological aspects of antioxidants used as food additives. In Food Antioxidants. Hudson, B. J. F., (ed.) Elsevier, London. 253-307. The histopathological changes of the aorta and liver support the previous study that Strobilanthes crispus plant may possess the hypocholesterolemic properties that can slower down the development of atherosclerosis.