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2007 - 200912010 - 20171
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Author: SRINIVASA NAIK, H × Start date: 2000 ×
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    ThesisItemOpen Access
    MOLECULAR AND PATHOLOGICAL STUDIES OF INDUCED HYPERLIPIDEMIA AND ATHEROSCLEROSIS IN RATS AND ITS AMELIORATION WITH FLAXSEEDS (LINUM USITATISSIUMUM) AND GREEN TEA (CAMELLIA SINENSIS)
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-05) SRINIVASA NAIK, H; SRILATHA, Ch(MAJOR); SUJATHA, K; SREEDEVI, B; PRASAD, T.N.V.K.V
      ABSTRACT : Hyerlipidemia is the disorder of lipid metabolism, characterized by elevated serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) and very low density lipoproteins cholesterol (VLDL-C) and decreased high density lipoprotein cholesterol (HDL-C). Atherosclerosis is diverse disease with heterogenous mechanisms of progression and is an oxidative, chronic inflammatory and thrombotic disease referring to fatty deposits on the inner lining of the large to medium sized elastic and muscular arteries and is precipitated by elevated level of low density lipoprotein cholesterol (LDL-C) in the blood. Atherosclerosis associated coronary heart disease is the second largest cause of mortality worldwide in humans. Current reports suggest that by the year 2020, India will have the largest cardio vascular disease (CVD) burden in the world. Realizing the importance of ethno medical practices, flaxseed and green tea supplementation along with atherogenic diet has been taken to combat the hyperlipidemia associated atherosclerosis. Flaxseed (Linumusitatissiumum) has been identified as a significant alternative source of omega 3 fatty acids and alpha linoleic acid (ALA). Therapeutic activities of flaxseed is well proven, one of them is anti-atherosclerotic due to its lipid lowering activity which is mainly attributed to its Secoisoarciresinol diglucoside (SDG). Green tea (Camellia sinensis) polyphenols known to have various medical health beneficial effects like anti-inflammatory, anti-oxidative, anti-arthritic, anti angiogenic, anti-metastatic, anti-cancer, anti-obesity, anti hyperlipidemic, anti-atherosclerotic, neuroprotective, anti-dental caries and antimicrobial (bacterial, viral & fungal) properties in human, animal and in vitro studies. So the present study was designed with a view to study molecular aspects of hyperlipidemia associated atherosclerosis in male Wistar albino rats and an attempt was made to identify various molecular preventive effects of flax seeds (Linumusitatissiumum) and green tea (Cameliasinensis). The present study was carried out by procuring 72 wistar albino rats, that were randomly divided into six groups consisting of 12 rats in each group. Hyperlipidemia associated atherosclerosis was induced by atherogenic diet consisting of 1% cholesterol and 15% saturated oil added to the 1000g of standard rat diet and given to group II rats. Flaxseed @ 7.5g/kg/day and green tea @ 100mg/kg/day gavaged orally to group V and VI rats for 90 days along with atherogenic diet. Group I kept as control and given standard rat diet, group III as flaxseed control and given 7.5g/kg/day of flaxseed along with standard rat diet, whereas group IV kept as green tea control and given 100mg/kg/day/Po along with standard rat diet for 90 days. Six rats from each group were sacrificed 45 days apart each. Atherogenic diet fed group (Group II) rats clinically showed obesity with significant increase (p<0.05) in the body weight. Rats were sluggish with poor hair coat. Green tea supplementation (Group VI) significantly reduced the obesity and body weight compared to moderate reduction of obesity and body weight in flaxseed ameliorated group (Group V). Whereas green tea control group (Group IV) rats were apparently healthy, slim with shiny hair coat. Total leukocyte count (TLC), total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C) were significantly (p<0.05) higher, whereas high density lipoprotein cholesterol (HDL-C) was significantly reduced in group II. Oxidative damage indicators like catalase, SOD, GPx, reduced glutathione and glutathione S transferase were significantly (p < 0.05) reduced, whereas lipid peroxidation products of TBARS level was non-significantly (p < 0.05) increased in liver and heart tissues of atherogenic diet fed group (Group II). Flaxseeds supplementation (Group V) reduced the hyperlipidemia certain extent caused by atherogenic diet by reducing the modest level of TC, TG, LDL-C and mild elevation of HDL-C. Flaxseeds non-significantly (p < 0.05) increased the cellular anti-oxidant enzymes and modestly reduced the level of TBARS. Whereas green tea (Group VI) supplementation effectively controlled the hyperlipidemia by reducing all the serum total cholesterol, triglycerides, low density lipoprotein cholesterol and very low density lipoprotein cholesterol and enhanced the good high density cholesterol and significantly (p < 0.05) enhanced all the tissue anti-oxidant enzymes and modestly reduced the TBARS levels. Microscopically hepatic steatosis was prominent in atherogenic diet fed group (Group II). Complete absence of fatty change was evident in green tea treated group (Group VI) compared to flaxseed group (Group V). Histopathologically, endothelial degeneration, sub intimal fat cells, foam cells with attached microthrombi was observed in majority of the cases in group II rats aorta fed on atherogenic diet. All these changes were also evidenced by ultrastructural studies. Mild endothelial degeneration was evident in flaxseed ameliorated group. Majority of the changes of initiated atherosclerosis were minimized by green tea supplementation. No such lesions were seen in control, flaxseed and green tea alone fed control groups (Group I, III & IV) throughout the study period. Mild endothelial cell proliferation with formation of vascular channels in between cardiac muscle fibers, liver and in kidney was observed throughout the study period of flaxseed fed group (Group III) rats. VEGF positivity was observed in endothelial cells, thrombus and structurally modified stromal cells of initiated atherosclerotic lesion. Initiated atheromatous plaque showed high level of positivity with Bax compared with other parts of blood vessel. On the other hand, selective moderate to severe positivity with CD 31, CD44, E cadherin and Beta catenin was observed in the initiated atherosclerotic lesion characterized by endothelial degeneration, sub endothelial lipid laden macrophages, and endothelial adhered thrombus and erythrocytes. Various levels of positivity with above markers was also observed in flaxseed and green tea ameliorated groups based on the initiated atherosclerotic lesion in these groups. Present study revealed positive expression of proinflammatory cytokine TNF alpha by RT-PCR (Real time PCR) in atherogenic diet fed group aorta (Group II) compared to standard diet fed rats (Group I). Flaxseed and green tea ameliorated groups (Group V and VI) showed significant reduction in the expression of TNF alpha in contrary to atherogenic diet fed group (Group II). IL-18 expression was not noticed in any groups of the present study. In conclusion atherogenic diet of present study established the hyperlipidemia and initiated the atherosclerosis in the aorta evidenced by histopathology and molecular expression of PECAM-I (CD 31), CD44, E cadherin and Beta catenin and tissue TNF alpha. Flaxseeds supplementation reduced the hyperlipidemia associated atherosclerosis to certain extent, but not completely ameliorated the changes. On the other hand, green tea supplementation effectively controlled the hyperlipidemia associated atherosclerosis and enhanced all the tissue anti-oxidant enzymes.
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    ThesisItemOpen Access
    STUDIES ON PATHOLOGY OF SHEEP LUNG
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-01) SRINIVASA NAIK, H; SRILATHA, Ch(MAJOR); SUJATHA, K; SURESH KUMAR, R.V
    ABSTRACT: In a developing agriculture based country like ours more than 70 crores of people are solely depend on agriculture and livestock and contributing more than 20% of G.D.P. annually. India ranks 6" among the countries of world with a population of 138.50 million sheep and Andrapradesh ranks IS' place in India with a sheep population of 21.37 millions. Lung infections are common in sheep, causing a great threat to the sheep rearing community. As disease problems severely cripples the production, a sound knowledge of common contagious or infectious diseases that accounts for morbidity and mortality in sheep become necessity. Keeping this in view, the present study on 'Studies on pathology of sheep lung' was undertaken to investigate various lung infections and isolation of bacteria those cause pneumonia in sheep and to know the ultra structural changes of pulmonary adenomatosis. Incidence of various types of pneumonia encountered were due bronchopneumonia (28%), pulmonary adenomatosis (9.8%), suppurative pneumonia (8.3%), interstial pneumonia (7.8%), fibrinous pneumonia (7.3%), Maedi (5.3%), pleuropneumonia (2.2%), hydatid cyst (1.26%), Sheep pox (1.0%), Tuberculosis (1 .OO/o), and neoplasm's like hemangioma and adenocarcinoma, noticed in two cases with an incidence of (0.5%) Bronchopneumonia was characterized by presence of moderate to severe infiltration of inflammatory cells, neutrophils and mononuclear cells in and around bronchioles and alveoli. Inter alveolar hemorrhages with mild edema of alveoli and peribronchial hemorrhages were seen. Pulmonary adenomatosis was characterized by papillary projections of epithelium into alveolar/ bronchiolar lumen. Fibrous tissue proliferation was seen. Metaplasia of alveolar epithelium leading to formation of glandular structures in alveoli was also evident. Suppurative pneumonia was characterized by areas of congestion, necrosis with calcification and presence of diffuse infiltration of inflammatory cells, mostly neutrophils and alveolar macrophages. Fibrous tissue proliferation was also evident. Interstitial pneumonia was characterized by thickened alveolar septa due to accumulation of serous and fibrinous exudates with infiltration of inflammatory cells like lymphocytes, macrophages and mononuclear cells in alveolar septa. In fibrinous pneumonia alveoli, bronchioles and bronchi were filled with sero fibrinous exudate. Infiltration of inflammatory cells like neutrophils and macrophages were seen along with edema, congestion and hemorrhages. Edema and widening of inter lobular septa was noticed in almost all cases with thickening of pleura. Maedi was characterized by hypertrophy and hyperplasia of bronchiolar epithelium. Peribronchiolar and perivascular infiltration with lymphocytes was most predominantly seen. Alveolar lumen contained desquamated epithelial cells. A thickened alveolar wall with interalveolar hemorrhages were also evident. Pleuro pneumonia was characterized by active hyperemia edema, congestion and hemorrhages with mild serous exudates along with mild infiltration of inflammatory cells like rnacrophages, lymphocytes and epitheloid cells. The pleura was thickened with fibrous tissue proliferation. Infiltration was very mild with neutrophiis and lymphocytes Tuberculosis was characterized by a granuloma with a central caseation and calcification surrounded by macrophages, epitheloid cells, lymphocytes, and Langhan's giant cells surrounded by fibrous covering. Other changes noticed were thickened pleura, sub pleural hemorrhages, pleural fibrosis and congestion. Hydatidosis was characterized by presence of mononuclear cells, eosinophils and plasma cell infiltration around the cystic space and surrounded by focal areas of fibrous tissue proliferation along with hemorrhages and congestion. Sheep pox was characterized by coagulative necrosis of the lobular tissue and infiltrated with the inflammatory cells mainly 'sheep pox' cells and macrophages and presence of serous exudate. The adjacent lung tissue showed congestion and infiltration with lymphocytes, neutrophils and macrophages. Neoplasms like hemangioma and adenocarcinoma were noticed in one case each. General pathological conditions encountered were pleuritis 2 (0.5%), bronchitis and bronchiolitis 15 (3.7%), bronchiectasis 2 (0.5%), bronchiolitis fibrosa oblitrans 2 (0.5%), pulmonary congestion and hemorrhages 17 (4.2%), pulmonary emphysema and atelectasis 16 (4.5%), pulmonary edema 16 (4.5%), pulmonary infarction 1 (0.2%), and anthracosis 1 (0.2%). Pleuritis was characterized by infiltration of inflammatory cells like neutophils and lymphocytes in the pleura and thickening of the pleura by edematous fluid. Bronchitis and bronchiolitis were characterized by presence of inflammatory cells like neutrophils, mononuclear cells and erythrocytes in the bronchi and bronchioles and these conditions were usually associated with bronchopneumonia. Widened lumen of bronchi contained mucus, large number of inflammatory cells, few erythrocytes. Bronchiolar wall was very thin and surrounded by fibrous tissue proliferation in bronchiectasis cases. Bronchiolitis fibrosa oblitranse was characterized by polypoid projections of bronchiolar epithelial and fibroblastic tissue, which was partially or completely, obliterated the lumen of bronchioles. Hemomes in the alveolar and bronchial spaces were noticed in pulmonary congestion and hemorrhages. Pulmonary emphysema was characterized by distended alveoli, widened interalveolar septa; few ruptured alveoli and atelectatic areas were characterized by cleft like alveoli with narrow lumen. Pulmonary edema was characterized by presence of edema fluid in the alveoli, interstitial tissue, interlobular septa and sub pleural zones and presence of compensatory emphysema and atelectatic changes. Pulmonary infarction revealed wedge or cone shaped hemorrhagic areas and presence of congestion, edema and atelectasis. Anthracosis was characterized by presence of black granules mostly in the alveolar walls and connective tissue septa of lung. In the present study 44 bacteria were isolated fiom 57 pneumonic ovine lungs. Pasteurella sps 10 (22.7%), E.coli 7 (1 5.9%), Corynebacterium sps 6 (13.6%), and Staphylococcus sps 6 (13.6%), were the major bacteria isolated. Stray cases of isolates were Streptococcus 3 (6.8%), Bacillus sps 2 (4.5%), Klebsiella sps 2 (4.5%), Proteus sps 2 (4.5%), Mjcobacterium 4 (9.0%), and Pseudomomis sps 2 (4.5%) cases. Major bacteria isolated fkom bronchopneumonia were Pastewella sps, followed by E-coli, Corynebacterium, and StaphyIococcal sps and from suppurative and fibrinous pneumonia cases, PasteureZla, E-coli. Corynebacterium were isolated. From pulmonary carcinoma, E. coli, Staphylococcus, Streptococcus was isolated and mycobacterium was identified in tuberculosis lung. Ultrastructurally, in all the cases of pulmonary adenomatosis proliferation of type I1 pneumocytes having microvilli, well-developed junctional complexes were noticed prominently. Mitochondria and endoplasmic reticulum were more numerous in type I1 penumocytes. In most of the cases mitochondria were well preserved with prominent cristae. Ciliated bronchiolar (Clara) cells with electron dense cytoplasmic granules without any surrounding membrane and microvilli were noticed.