ASIAN J. EXP. BIOL. SCI.

VOL 3 (1) 2012: 197 - 202

Society of Applied Sciences

ORIGINAL ARTICLE

Risk of CAD in Urban and Rural Diabetic Population

1

M.Saraswati, G.Rajesh Kumar, K.Mrudula Spurthi, G.Srilatha, P.Chiranjeevi, Vijay Sekhar 1 Reddy and H. Surekha Rani
1

Dept. of Genetics, University College of Science, Osmania University, Hyderabad, 500007, Andhra Pradesh, India. 2 Department of Endocrinology, Gandhi Hospital, Secunderbad, 500003, Andhra Pradesh, India. . ABSTRACT Type 2 diabetes is increasing at an alarming rate all over the world. It is a disorder characterized by hyperglycemia and associated with microvascular and macrovascular complications. Among macrovascular complications, coronary artery disease (CAD) has been associated with diabetes in numerous studies. Oxidative stress due to hyperglycemia in diabetes contributes to the development of diabetic complications. Lipid peroxidation (malondialdehye-MDA) of cellular structures is important process in diabetes mellitus and its complications. Nitric oxide (NO) impairment is regarded as an early step in the development of insulin resistance, atherosclerosis, and type 2 diabetes. Rapid urbanization and industrialization together has also been a causative factor posing a growing threat to the health of the population. Hence, the present study has been designed to evaluate MDA an end product of lipid peroxidation along with nitrite/nitrate levels and lipid profiles as risk factors for coronary artery disease in type 2 diabetics in urban and rural population. The results of the present study demonstrated an increased level of oxidative stress markers and altered lipid profiles in urban diabetics and healthy controls corresponding to respective rural population suggesting the effect of urbanization and impact of different life styles on the health of the people. Key Words: Type 2 diabetes, Coronary artery disease, oxidative stress, urbanization

INTRODUCTION Type 2 diabetes mellitus is a polygenic and multifactorial disorder resulting from the interaction between a genetic predisposition, environmental risk factors and behavioral patterns [1, 2]. It comprises a series of dysfunctions resulting from the combination of resistance to insulin action to inadequate insulin secretion [3]. The incidence of type 2 diabetes mellitus is increasing worldwide and the prevalence of diabetes for all age-groups worldwide was estimated to be 2.8% in 2000 while recent trends show that it might increase to 4.4% in 2030 [4]. The number of people with diabetes is increasing due to population growth, aging, increasing prevalence of obesity, physical inactivity and urbanization. The difference in the prevalence of diabetes in urban and rural indicates a major role of urbanization in the occurrence of the type 2 diabetes [5]. Adults with diabetes mellitus also frequently develop micro and macro vascular complications which include neuropathy, nephropathy, cerebrovascular, cardiovascular diseases etc. [6].There is an enhanced risk of coronary artery disease (CAD) morbidity and mortality in type 2 diabetic patients [7-9]. Elevated levels of serum LDL cholesterol, triglyceride and low levels of HDL cholesterol have been reported as most important risk factors for CAD [10-13]. Increasing evidence in both experimental and clinical studies suggest that oxidative stress due to hyperglycaemia plays an important role in the pathogenesis of type 2 diabetes mellitus and its complications [14]. Hence, the present study has been designed to evaluate the oxidative stress markers (MDA-malondialdehye, nitrite /nitrate) and lipid profiles as risk factors for coronary artery disease in type 2 diabetics in urban and rural population.

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012

197

Risk of CAD in Urban and Rural Diabetic Population.............................................................................................................................M.Saraswati et al.

MATERIALSAND METHODS Study population and design: After careful clinical examination and confirmed diagnosis by endocrinologist , 100 patients (urban and rural) presenting type 2 diabetes along with 100 age and gender matched healthy controls (urban and rural) were included in the study. The biochemical investigations were carried out in the blood sample of patients and healthy controls. Total cholesterol, HDL-cholesterol and triglycerides were estimated by using commercially available kits and measured with SHIMADZU UV 240-Spectrophotometer and LDL-cholesterol, was calculated according to the Friedewald equation: VLDL = Triglycerides/5 LDL = Total cholesterol (HDL + VLDL). Lipid peroxidation (MDA): Estimation of plasma malondialdehyde (MDA) was carried out by Gavino et al method: to 0.5ml of freshly obtained plasma an equal volume of 0.9% saline and trichloroacetic acid (TCA) was added and incubated at 370c for 20 minutes and centrifuged for 10min at 3000rpm and to 1ml of protein free supernatant 0.25ml 0 of thiobarbituric acid (TBA) was added and incubated for 60min at 95 c and measured with SHIMADZU UV 240Spectrophotometer [15]. Estimation of nitrite / nitrate: Nitrite / Nitrate concentrations present in the reaction mixture were determined by using Griess reagent (a 1:1 mixture of 1% sulfanilamide in 5% H3PO4 and 0.1 % N-(1-napthyl)-ethylene-diamine ) by Lepovire et al method : 0.5 ml of serum was precipitated with 50 l of 70% sulfosalicylicacid (SSA), mixed well for 5 minutes, vortexed and then centrifuged at 3000 rpm for 20 min. 200 l of supernatant was taken and 30 l of 10% NaOH, 300 l of 50 mMtris buffer and 530l of Greiss reagent were added and incubated for 10 min in dark. The absorbance was read against blank (double distilled H20) at 540 nm using SHIMADZU UV- 240 spectrophotometer. The concentration of nitrite/ nitrate in serum was determined based on the standard curve generated [16]. RESULTS Significant differences were observed between the groups under study. Lipid profiles were significantly different in diabetic urban and rural patients compared to controls .Mean and SD of MDA and nitrate/nitrite levels in the patients of urban and rural were significantly different when compared to controls. Lipid profiles of controls and patients are presented in table 1:
TABLE 1: Serum lipid profiles of healthy controls and diabetic patients.

Subjects

Total Cholesterol mg% mean SD 129 27.0 145 31.0

LDL Cholesterol mg/dl mean SD 90 8.1 95 8.8

HDL Cholesterol mg/dl mean SD 45 10.05 43.25 12.15

Triglycerides mg/dl mean SD 141 14.75 152 17.23

Controls [n=100] Rural:50 Urban:50 Diabetics [n=100] Rural:50 Urban:50

191 29.3* 242 30.3*

133 32.50* 148 38.06*

38 5.1 30 4.5

181 25.57* 195 16.54*

*Significantly higher at p < 0.01.

198

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012

Risk of CAD in Urban and Rural Diabetic Population.............................................................................................................................M.Saraswati et al.

The mean SD levels of total cholesterol, LDL-cholesterol and triglycerides in the diabetic patients (urban/ rural) were significantly high at p<0.01 compared to respective healthy controls (urban/ rural). The mean SD levels of the total cholesterol, LDL-cholesterol and triglycerides in the urban diabetic patients were found to be 242 30.3, 148 38.06, 195 16.54 and that of the urban controls were 145 31.0 , 95 8.8 , 152 17.23 significant at p<0.01 while the mean SD levels of the total cholesterol, LDL-cholesterol, triglycerides in the rural diabetic patients were found to be 191 29.3 , 133 32.50 , 181 25.57 and that of the controls were 129 27, 90 8.1 , 141 14.75 significant at p<0.01 . The mean SD levels of HDL-cholesterol in the diabetic patients (urban/ rural) were significantly low at p<0.01 compared to respective healthy controls (urban/ rural). The mean SD levels of the HDL-cholesterol, in the urban diabetic patients were found to be 30 4.5 and that of the urban controls were 43.25 12.15 significant at p<0.01 while the mean SD levels of the HDL-cholesterol in the rural diabetic patients were found to be 38 5.1 and that of the controls were 45 10.05 significant at p<0.01 . Oxidative stress markers (MDAand nitrite/nitrate) of patients and controls are presented in table 2:
TABLE 2: Markers of oxidative stress of healthy controls and diabetic patients.

Subjects

MDA levels [nmoles/ml] Mean SD

Nitrite/Nitrate levels [Moles/ml] mean SD] 1.21 0.52 1.68 0.36 2.01 0.15* 3.13 0 .98*

Controls[n=100] Rural:50 Urban:50 1.37 0.30 1.58 0.57 1.98 0.57* 2.75 0.64*

Diabetics [n=100] Rural:50 Urban:50

*Significantly higher at p < 0.01.

The mean SD levels of MDA in the diabetic patients (urban/ rural) were significantly high at p<0.01 compared to respective healthy controls (urban/ rural). The mean SD levels of MDA , in the urban diabetic patients were found to be 2.75 0.64 and that of the urban controls were 1.58 0.57 significant at p<0.01 while the mean SD levels of the MDAin the rural diabetic patients were found to be 1.98 0.57 and that of the rural controls were 1.37 0.30 significant at p<0.01 . The mean SD levels of nitrite/nitrate in the diabetic patients (urban/ rural) were significantly high at p<0.01 compared to respective healthy controls (urban/ rural). The mean SD levels of nitrite/nitrate, in the urban diabetic patients were found to be 3.13 0.98 and that of the urban controls were 1.68 0.36 significant at p<0.01 while the mean SD levels of the nitrite/nitrate in the rural diabetic patients were found to be 2.01 0.15 and that of the rural controls were 1.21 0.52 significant at p<0.01. Study markers in urban and rural diabetics: The mean SD levels of total cholesterol, LDL-cholesterol, triglycerides, MDA and nitrite/nitrate were high while mean SD levels of HDL- cholesterol were low in the urban diabetics significant at p<0.01 compared to rural diabetics. Study markers in urban and rural healthy controls: The mean SD levels of total cholesterol, LDL-cholesterol, triglycerides, MDA and Nitrate/Nitrite were high while mean SD levels of HDL- cholesterol were low in the urban healthy controls significant at p<0.01 compared to rural healthy controls.

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012

199

Risk of CAD in Urban and Rural Diabetic Population.............................................................................................................................M.Saraswati et al.

DISCUSSION st Diabetes is now widely regarded as the 21 century epidemic. The prevalence of the disease has risen substantially over the past 10 years and is a major emerging clinical and public health problem. It has been reported that worldwide approximately 284 million people are currently diagnosed with the disease. As the world becomes increasingly urban and megacities emerge, traffic related air pollution poses environmental, ecological and human health risks. Rural populations with more traditional life styles exhibit lower rate of diabetic risk factors and diabetes, whereas urban populations and particularly those of a higher socioeconomic status have higher rates of both risk factors and diabetes [17]. Presently the data on urban and rural populations and type 2 diabetes risk and its complications are limited. Coronary artery disease (CAD) is more prevalent among subjects with type 2 diabetes mellitus, than compared to the other population. Higher risk of CAD is seen in diabetics characterized by lipid abnormalities in having decreased HDL cholesterol and hypertriglyceridemia. Similarly various studies have suggested an association of lipid abnormalities and increased risk of CAD thus leading to higher mortality rate in type 2 diabetics. [18-22]. The estimation of lipid profiles in the diabetics serves to monitor the prognosis of the subject for detection of risk factors in the early stage of the disease which would then help to improve and reduce rate of morbidity [23].Total and LDL cholesterol levels in diabetics are comparable with levels found in non-diabetics while low levels of HDL cholesterol and elevated TG levels, both contributing factors for cardio-vascular diseases, have been reported in type 2 diabetes [24, 25]. Therefore the present study has evaluated the risk factors for coronary artery disease in urban and rural diabetic population in comparison to the respective healthy controls. Our study had similar observations and found higher levels of cholesterol and triglycerides and lower levels of HDL cholesterol in diabetics compared to the corresponding healthy controls. We have further observed that higher levels of triglycerides and lower levels of HDL cholesterol in urban diabetic population compared to rural diabetic population as shown in table 1, suggests the risk of CAD in urban compared to rural diabetics. Another alarming observation is that the urban healthy controls had higher values of cholesterol and triglycerides and lower values of HDL cholesterol when compared to the rural healthy control population as depicted in table 1, indicating the future risk of association of health problems with different lifestyles, environmental factors etc. Further the higher levels of cholesterol, triglyceride, LDL-cholesterol in urban population may be attributed to high calorie intake, unhealthy diets, lack of exercise, obesity, sedentary life style in the patients of diabetes mellitus [26, 27]. Numerous epidemiological studies have also shown a higher prevalence of type 2 diabetes in urban areas compared with rural areas [28, 29]. This difference has been observed particularly in developing countries that have undergone a rapid transition from rural to urban lifestyle. It has been largely attributed to broad shifts in diet, physical activity, and obesity in urban areas. However, these lifestyle factors fail to completely explain the association with increased diabetes and obesity risk [30]. Hyperglycemia in diabetic condition depletes natural antioxidants and facilitates the production of reactive oxygen species-ROS which has the ability to react with all biological molecules. Thus increased ROS and impaired antioxidant defense contributes for initiation and progression of micro and macro vascular complications in diabetics like, coronary artery diseases, diabetes nephropathy, retinopathy, neuropathy etc. [31]. Oxidative stress can be measured by monitoring blood malondialdehyde (MDA) which is a marker of lipid oxidation. It has been suggested that MDA an end product of lipid peroxidation, plays a key role in the development of atherosclerosis [32]. De Zwart et al (1999) proposed that oxidative stress may be associated with the pathogenesis of non-insulin dependent diabetes mellitus complications, particularly cardiovascular diseases [33]. In the present study MDA levels were significantly higher in the diabetics compared to the healthy controls. A similar finding was reported by Dierckx et al, (2003) Hanachi et al (2009), Jain et al (1999). Thus oxidative stress can act as a contributor in the pathogenesis of cardiovascular diseases [34-37]. Hyperglycaemia a contributing factor in the progressive pathogenesis of diabetic complications is likely to be a causing factor to reduce bioavailability of NO [38]. Higher levels of NO promote the peroxidation of the lipid moiety and induce immune responses and inflammatory reactions that cause cell damage [39] .Apositive association between NO2 exposure as a marker of traffic related air pollution and type 2 diabetes has also been reported [40]. NO is a nonstable radical and converted to nitrite/nitrate anion (NO2_/NO3_) in a very short time which is more stable product [41]. In the present study nitrite/nitrate levels were significantly higher in the diabetics compared to the healthy controls. We have observed higher levels of MDA and nitrite\nitrate in diabetics (urban and rural) compared to the
200

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012

Risk of CAD in Urban and Rural Diabetic Population.............................................................................................................................M.Saraswati et al.

corresponding healthy controls. We have further observed higher levels of MDA and NO (nitrite\nitrate) in urban diabetic population when compared to rural diabetic population as shown in table 2. Therefore the study suggests that increased levels of oxidative stress markers contribute to the atherosclerotic process that eventually leads to coronary artery disease in diabetic patients and also suggests a higher risk of complications in urban diabetics compared to rural diabetics. A link between pollution exposure and physiological responses has been suggested which are known to be along the pathway of adverse cardiovascular outcomes. Higher rates of cardiac hospitalization and mortality on high pollution days among people with diabetes may be partially explained by impairments in endothelial function, vascular smooth muscle function [42]. Exposure of diabetic individuals to exogenous sources of free radicals (traffic, industrial emissions etc.) in urban areas may further enhance the complications associated with diabetes. Whitsel et al (2009) reported that insulin and insulin resistance increase susceptibility to the adverse effect of pollution on cardiac autonomic control. Such increases in susceptibility may, in turn, influence the risk of coronary heart disease among persons with type 2 diabetes [43]. The study also further observed an increase of oxidative stress markers MDA and nitrite/nitrate in the urban healthy controls when compared to the rural healthy controls suggesting a higher risk of future health problems in urban population compared to rural population. Narasimha et al (2010) advocated that the estimation of lipid peroxide along with other lipid profiles in the type 2 diabetes may serve as a useful monitor to judge the prognosis of the patient [44]. As suggested by Ursula Kramer (2010) environmental factors would also be expected to contribute to these striking differences, and further studies are needed to investigate the extent to which pollution determines the environmental health burden in urban and rural areas [31].

CONCLUSION The present study demonstrated an increased level of oxidative stress markers (MDA, NO) and altered lipid profiles in urban (diabetics and healthy controls) corresponding to respective rural population suggesting the effect of environmental pollution and different life styles on the health of the people. Urbanization and climate change works synergistically to increase the disease burdens. Hence newer strategies have to be designed to reduce the disease burden of type 2 diabetes and its complications like CAD. REFERENCES
[1]. Tuomilehto,J.,Lindstrm,J.,Eriksson,J.G., Valle,T.T., Hmlinen,H., Ilanne,P.P., Keinne N, S., Laakso,M.L., Ouheranta, A., Rastas,M., Salminen,V and Uusitupa, M (2001). Prevention of Type 2 Diabetes Mellitus by Changes in Lifestyle among Subjects with Impaired Glucose Tolerance. N Engl J Med; 3: 344(18):1343-50. [2]. James,V.N (1962). Diabetes Mellitus:AThrifty Genotype Rendered Detrimental by Progress? Am J Hum Genet.; 14(4): 35362. [3]. Marios, K (2005). Target gene discovery in extended families with type 2 diabetes mellitus. Atherosclerosis supplements; 6(2):31-36. [4]. Jansson,S.P.O., Andersson, D.K.G and Svrdsudd, K (2007). Prevalence and incidence rate of diabetes mellitus in a Swedish community during 30 years of follow-up.Diabetologia; 50:70310. [5]. Wild,S., Roglic,G., Green,A., Sicree,R and King,H (2004). Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care; 27:104753. [6]. Pham-Huy, L.I., He,H and Pham-Huy, C (2008). Free radicals, antioxidants in disease and health. Int J Biomed Sci; 4(2): 89-96. [7]. Grundy,SM, Benjamin,I.J., Burke,G.L., Chait,A., Eckel,R.H., Howard,B.V. Mitch, W.,Smith, S.C.J r. and Sowers, J.R (1999). Diabetes and cardiovascular disease: a statement for health care professionals from the American Heart Association. Circulation: 100(10): 113446. [8]. Sundarajan,S and Prakash,D (2007). Management of coronary artery disease in patients with type 2 diabetes mellitus. Current Cardiology Reports; 9(4):264-71. [9]. Jeroen,J.B., Lawrence,H,Y., Robert,LF., Robert,O.B., Helmut,O,S and Eugene,J.B (2007). Screening for Coronary Artery Disease in Patients with Diabetes. Diabetes Care; 30 (10): 2729-36. [10] .Kolovou,G.D., Anagnostopoulou,K., Pilatis,N.D., Salpea,K.D., Hoursalas,I.S., Petropoulos,I., Bilianou, H.I and Cokkinos,D.V (2005). Fasting serum triglyceride and high-density lipoprotein cholesterol levels in patients intended to be treated for dyslipidemia. Vasc Health Risk Manag; 1(2):155-61. [11]. Shepherd,J (1998) Profiling risk and new therapeutic interventions: looking ahead. Am J Med.; 104(2A):19S-22S. [12]. LaRosa,J.C (1992). Lipids and cardiovascular disease: do the findings and therapy apply equally to men and women? Womens Health Issues; 2(2):102-11. [13]. Gordon,D.J (1990). Role of circulating high-density lipoprotein and triglycerides in coronary artery disease: risk and prevention. Endocrinol MetabClin North Am.; 19(2):299- 309. [14]. Mattia, D., Laurenti, G. O. and Fava, D (2003). Diabetic endothelial dysfunction: effect of free radical scavenging in Type 2 diabetic patients. J Diabetes Complications; 17(2 Suppl):30-5. [15].Gavino,V.C., Miller,J.S., Ikharebha,S.O., Milo,O.E and Cornwell, D.G (1981). Effects of polyunsaturated fatty acids and antioxidants

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012

201

Risk of CAD in Urban and Rural Diabetic Population.............................................................................................................................M.Saraswati et al.

on lipid peroxidation in tissue cultures Journal of Lipid Research ; 22:763-9. [16]. Lepoivre, M., Chenal, B., Vapo, A., Lamair, G., Thalander, L. and Tenu, J. P (1990). Alteration of ribonucleotidereductase activity following induction of nitrite generating pathway in adenocarcinoma cells. J. Biolchem; 265:14143-9. [17]. William,H.H., Ronald,E.A., Mohammad,A.A., Edward,S.S and Ahmed,B (1997). Diabetes mellitus in Egypt: risk factors, prevalence and future burden. Eastern Mediterranean Health Journal; 3(1): 144-8. [18]. Licia,L., Elisabetta,C and Maria,B.D (2002). Review: The genetics of macrovascular disease in diabetes. The British Journal of Diabetes & Vascular Disease 2:5: 364-8. [19]. Barrett,C.E., Grundy,S.M and Holdbrook, M.J (1982). Plasma lipids and diabetes mellitus in an adult community. Am. J. Epidemiol.; 115: 65763. [20]. Sowers, J.R and Lester. M.A(1999). Diabetes and cardiovascular disease. Diabetes Care; 22: 14-20. [21]. Laasko,M (1996). Lipids and lipoproteins as risk factors for coronary heart disease in noninsulin-dependent diabetes mellitus. Ann. Med; 28: 341-5. [22]. Miller, M (1999). The epidemiology of triglyceride as a coronary artery disease risk factor. Clin. Cardiol; 22(Suppl. II): II16. [23]. Suryawanshi, N.P., Bhutey, A.K., Nagdeote, A.N., Jadhav,A.A and Manoorkar,G.S (2006).Study of lipid peroxide and lipid profile in diabetes mellitus. Indian Journal of Clinical Biochemistry; 21 (1) 126-30. [24]. Alireza,E., Mehrshad,A., Manouchehr,N., Abbas,Y., Amelita,P.B and Hamid,A (2006). Prevalence of diabetes and other cardiovascular risk factors in an Iranian population with acute coronary syndrome. Cardiovascular Diabetology ; 5:15: 1-6. [25]. Richard,W.N (2008). LDL Cholesterol Lowering in Type Diabetes: What Is the Optimum Approach? Clinical Diabetes; 26 (1):8-12. [26]. Siddartha,D., Samal,K.C and Tripathy,B.B (1992). Factors influencing plasma lipids and lipoprotein cholesterol in Indian NIDDM. J. Dia. Assoc. Ind. 32 (2). [27]. Analava,M., Bhattacharya, D and Roy, S (2007). Dietary influence on TYPE 2 Diabetes (NIDDM). J. Hum. Ecol.; 21(2): 139-147. [28]. Ramachandran, A., Simon,M., Annasami,Y., Narayanasamy,M and Chamukuttan, S (2008).High prevalence of diabetes and cardiovascular risk factors associated with urbanization in India. Diabetes care; 31(5), 893-8. [29]. Sobngwi, E., Mbanya,J.C., Unwin,N.C., Porcher,R., Kengne,A.P and Fezeu,L (2004). Exposure over the life course to an urban environment and its relation with obesity, diabetes, and hypertension in rural and urban Cameroon. Int J Epidemiol; 33:76976. [30]. Barry,M.P (1999) Urbanization, lifestyle changes and the nutrition transition. World Dev; 27:190516. [31]. Ursula,K., Christian,H., Dorothea,S., Klaus,S., Tamara,S., Ulrich,R and Wolfgang,R (2010). Traffic-related air pollution and incident type 2 diabetes: Results from the SALIACohort study. Environmental Health Perspectives; 118(9):1273-79. [32]. Ayaz,K.M., Ravindra,M., Vivek,R.J., Gaurav,S. & Marya,A (2011). A study on malondialdehyde as a marker of lipid peroxidation in male and female patients with type 2 diabetes mellitus.International Journal of Pharmaceutical Sciences Review and Research; 8 (2)33:198-201. [33]. De Zwart,L,L., Meerman,J.H., Commandeur,L.N and Vermeulen,N.P (1999). Biomarkers of free radical damage applications in experimental animals and in humans. Free Radical Biology and Medicine, 26: 202-26. [34]. Dierckx,N., Horvath,G., Van,G.C., Vertommen,J., Van De,V. J., De Leeuw, I and Manuel,Y. K.B(2003). Oxidative stress status in patients with diabetes mellitus: relationship to diet. Eur J ClinNutr ; 57(8):999-1008. [35]. Hanachi,P., Rashid,H.M. &Latiffah,A.L (2009). Investigation of Lipid Profiles and Lipid Peroxidation in Patients with Type 2 Diabetes European Journal of Scientific Research; 28 (1): 6-13. [36]. Jain,S.K., McVie,R., Jackson,R., Levine,S.N and Lim,G (1999). Effect of Hyperketonemia on Plasma Lipid Peroxidation Levels in Diabetic Patients. Diabetes Care; 22:1171-5. [37]. Saverio,S., Joan,M.D., Richard,P.D., Richard,W.B., Freudenheim Jo,L., Kathleen,M.H. & Maurizio,T (2008). Oxidation, Type 2 Diabetes, and Coronary Heart Disease:AComplex Interaction Findings from a population-based study. Diabetes Care; 31:18646. [38]. Francesco,C., Masato,E., Paolis,De,P., Bernd van der, L., Markus,B., Volker,U., Alexei,K., Chiara,D.G., Hana, J., Massimo,V and Thomas,F.L ( 2003) . High glucose causes upregulation of cyclooxygenase-2 and alters prostanoid profile in human endothelial cells role of protein kinase C and reactive oxygen species. Circulation ;107(7):1017-23. [39]. Leeuwenbergh,C., Hardy, M.M., Hazen,S.L., Wagner,P., Oh- Ishi,S., Steinbrecher,U.P and Heineck, J.W (1997). Reactive nitrogen intermediates promote low density lipoprotein oxidation in human atherosclerotic intima. J. Bio.Chem ; 272: 1433-6. [40]. Robert,D.B., Barry,F., Wayne,C., Yuling,H., George,H., Michael,L., Russell,L., Murray,M., Jonathan,S., Sidney,C.S and Jr Ira T (2004).Air pollution and cardiovascular disease. Circulation; 109; 2655-71. [41]. Aydin,A., Orhan,H., Sayal,A., zata,M., Sahin,G.& Isimer, A (2001). Oxidative stress and nitric oxide related parameters in type II diabetes mellitus: effects of glycemic control.ClinicalBiochemistry; 34, (1), 65-70. [42]. Marie,S.O'N., Aristidis,V., Antonella, Z., Jeremy,A.S., Diane,R.G., Panayiotis,A.E., Edward, S. H. and Joel, S (2005).Diabetes enhances vulnerability to particulate air pollutionassociated impairment in vascular reactivity and endothelial function; Circulation; 111:2913-20. [43]. Whitsel,E.A., Quibrera,P.M., Christ,S.L., Liao,D., Prineas,R.J and Anderson,G (2009). Heart rate variability, ambient particulate matter air pollution, and glucose homeostasis: the environmental epidemiology of arrhythmogenesis in the women's health initiative. Am J Epidemiol; 169:693703. [44]. Narasimha, R.K and Jeganathan P.S (2010). A correlative study of status of serum cholesterol,triglyceride and HDL-cholesterol in Type 2 diabetes mellitus patients with and without hypertension. Research Journal of Pharmaceutical, Biological and Chemical Sciences; 1(3):377-81.

Correspondence to Author : H. Surekha Rani ,Dept. of Genetics, University College of Science, Osmania University, Hyderabad, 500007, Andhra Pradesh, India. Email: surekharanih@gmail.com
202

ASIAN J. EXP. BIOL. SCI. VOL 3 (1) 2012