Effect of Succinamic Acid Derivative on the Growth Kinetics and Induction of Apoptosis in a Cancer Cell Line.

Introduction Head and neck cancers are heterogeneous malignancies associated with significant morbidity. Oral cancers are related to the use of tobacco products. Smokeless tobacco usage is a health problem worldwide, and its carcinogenic mechanism is largely unknown. Despite advances in conventional treatments, side effects and drug resistance remain unsolved. Therefore, novel therapeutic agents with minimal side effects using plant derivatives should be explored. An active antihyperglycemic and antioxidant compound known as FIIc was isolated from the fruit pulp of Eugenia jambolana (US Patent No.: 2,30,753). Although E. jambolana is reported to have anticancer activity, no study has been reported on its growth kinetics and apoptotic potential in the human head and neck cancer cell line (SCC4). The present study evaluated the effect of an herbal compound isolated from the fruit pulp of E. jambolana and chemically synthesized the same compound, α-hydroxy succinamic acid (α-HSA), on SCC4 proliferation and apoptotic gene expression. Methods The SCC4 cell line was cultured in Dulbecco's Modified Eagle Medium (DMEM). The dosages of smokeless tobacco extract (STE), herbal compound, and synthetic compound were determined by cell viability assay, and their effect on mRNA expression of apoptotic genes was measured by real-time polymerase chain reaction. Results The present study observed significant therapeutic effects of the natural and synthetic compounds from the fruit pulp of E. jambolana at the concentration range of 100-200 µg/mL on the SCC4 cell line. α-HSA had antiproliferative action; upregulated apoptotic genes like p53, p21, and Bax; and downregulated anti-apoptotic genes like survivin in the SCC4 cell line. Conclusion The therapeutic potential of α-HSA and the putative mechanisms involved may be explored to provide the basis for future therapeutic interventions in oral cancer mediated by smokeless tobacco.

Association of oxidative stress and inflammatory markers with chronic stress in patients with newly diagnosed type 2 diabetes.

AIMS: Chronic stress is associated with increased risk of type 2 diabetes. Oxidative stress and inflammation are potential mediators of this risk. This study was conducted to investigate the association of oxidative stress and inflammatory markers with chronic stress and newly diagnosed type 2 diabetes. METHODS: Oxidative stress/antioxidant status (malondialdehyde [MDA], reduce glutathione [GSH], glutathione reductase [GR], glutathione peroxidase [GPx], catalase [CAT], superoxide dismutase [SOD]), inflammatory markers (highly sensitive C-reactive protein [hsCRP], adiponectin, leptin), chronic stress levels as assessed by stress scales-presumptive stressful life events scale (PSLES), perceived stress scale (PSS), sense of coherence (SOC) and stress biomarker-salivary cortisol in 125 subjects with newly detected diabetes mellitus (NDDM) were compared with an equal number of age and sex matched subjects with normal glucose tolerance (NGT). RESULTS: NDDM subjects as compared with NGT had significantly increased MDA (P < 0.001), hsCRP (P < 0.001), and leptin (P = 0.014) levels and increased GR (P = 0.043) and SOD (P < 0.001) activity along with decreased GSH (P < 0.001) and adiponectin (P < 0.001) levels. They also had significantly higher PSLES-LT and PSS and lower SOC scores along with elevated levels of 10:00 pm salivary cortisol and post dexamethasone salivary cortisol as compared with NGT. In stepwise logistic regression analysis, variables GSH (OR: 0.805), SOD (OR: 1.004), and adiponectin (OR: 0.771) along with PSLES-LT (OR: 1.007), PSS (OR: 1.147), SOC (OR: 0.667), salivary cortisol 10:00 pm (OR: 1.382), WC (OR: 1.087), and HOMA-IR (OR: 2.721) emerged as significant predictors of NDDM. CONCLUSION: The findings of this study indicate that chronic psychological stress and stress responses are associated significantly with inflammation and oxidative stress, which could act as mediating stress related risk factors for type 2 diabetes.

Glycated apolipoprotein B-A surrogate marker of subclinical atherosclerosis.

AIMS: Sustained hyperglycemia is a causative factor for glycation of proteins. Glycated low-density lipoprotein (LDL) is strongly associated with an increased risk of CAD (Coronary Artery Disease) in diabetics. Hence, we planned to evaluate the association of glycated apo B with subclinical atherosclerosis. METHOD: Forty-five non obese and 45 obese diabetics were recruited. Glycated hemoglobin (HbA1c) levels were estimated by HPLC (High Pressure Liquid Chromatography) and small dense low-density lipoprotein (sdLDL) was calculated using standard formula. Plasma Insulin was done by RIA. Insulin resistance was calculated using homeostatic model assessment insulin resistance (HOMA-IR) model. Glycated apo B in serum was estimated using ELISA. Carotid intimal media thickness (CIMT) was estimated using B mode USG of carotid arteries. RESULTS: Glycated apo B levels were correlated significantly with fasting blood glucose (FBG) (p=0.001), post prandial glucose (PPG) (p=0.001), HbA1c (p=0.013). The percent glycated apo B levels correlated significantly with FBG (p=0.032), PPG (p=0.004) in obese diabetic group. Multivariate regression analysis of glycated apo B and percent glycated apo B, showed that glycated apo B (p=0.009) and percent glycated apo B (p=0.006) were significantly correlated to FPG in diabetic population. The percent glycated apo B was also significantly correlated to PPG (p=0.003) and sdLDL (p=0.009). CIMT levels were higher in obese diabetics with 2 plaques positive when compared to obese non diabetic controls; however levels were not statistically significant. CONCLUSION: Persistent hyperglycemia and sdLDL are independently associated with glycation of apo B. Presence of plaques and increased thickness of intima indicates that glycated apo B predisposes diabetics to atherosclerosis.

ß-Adrenoreceptor Agonist Isoproterenol Alters Oxidative Status, Inflammatory Signaling, Injury Markers and Apoptotic Cell Death in Myocardium of Rats.

Sustained high levels of circulating catecholamines are reported to induce cardiotoxicity. Isoproterenol (ISP), a synthetic catecholamine has been widely employed to induce myocardial injury, though the role of inflammation and apoptosis is not well established. This study was designed to investigate the underlying mechanism of oxidative damage, inflammatory signaling, cell death in ISP induced myocardial infarction in rats. Wistar albino rats were divided in two groups: group I (sham control) and group II (ischemic control). ISP (85 mg/kg, s.c.) was administered at an interval of 24 h to group II for two consecutive days. On day third, after 48 h of the first injection of ISP, blood was collected from retro orbital plexus of rat eyes to estimate the biochemical parameters. Glutathione (GSH) and superoxide dismutase (SOD) were measured for antioxidant status. Similarly, malondialdehyde (MDA) was measured as an index of lipid peroxidation. Cardiac markers (SGOT, CK-MB, TropI and LDH) and pro-inflammatory cytokines (IL-6, CRP and TNF-α) were also estimated in ISP-induced rats. At the end of experiments animals were sacrificed for histopathological studies. GSH and SOD showed significant decrease after ISP challenge as compared to sham (control) group (p < 0.01) while MDA level, increased significantly (p < 0.01). ISP, also increased the level of cardiac markers and markers of inflammation significantly (p < 0.01), which was further verified by histopathological studies of the heart tissues. The study confirmed that ISP causes detrimental changes in the myocardium by altering cardiac and inflammatory markers, which leads to severe necrosis. The deleterious effects produced by ISP substantiate its suitability as a novel animal model for evaluation of cardioprotective agents/drugs.

Terminalia arjuna (Roxb.) Wight & Arn. augments cardioprotection via antioxidant and antiapoptotic cascade in isoproterenol induced cardiotoxicity in rats.

Worldwide, Ischemic heart disease (IHD) affects a large population. Implication of myocardial infarction (MI) and its multiple pathophysiology in cardiac function is well known. Further, isoproterenol (ISP) is known to induce MI. Today, there is an urgent need for effective drug that could limit the myocardial injury. Therapeutic intervention with antioxidants has been shown useful in preventing the deleterious changes produced by ISP. Here, we investigated the protective effects of oral pre-treatment of hydroalcoholic extract of bark of Terminalia arjuna (HETA) on biochemical and apoptotic changes during cardiotoxicity induced by isoproterenol (ISP) in rats. HETA was orally administered at a dose of 100, 200 and 400 mg/kg body wt., for 30 days with concurrent administration of ISP (85 mg/kg body wt.) on days 28th and 29th at an interval of 24 h. ISP caused deleterious changes in the myocardium and significantly increased (P < 0.05) malondialdehyde, serum glutamate oxaloacitate transaminase, creatine kinase-MB, lactate dehydrogenase and troponin-I. However, it significantly decreased (P < 0.05) glutathione and superoxide dismutase compared to healthy control. Oral pre-treatment of HETA for 30 days significantly decreased (P < 0.05) the biochemical parameters of oxidative stress and cardiac markers as compared to ISP control. Histopathological findings also revealed that architecture of the myocardium was restored towards normal in HETA pre-treated group. Overall, the present study has shown that the hydroalcoholic extract of bark of T. arjuna (HETA) attenuates oxidative stress, apoptosis and improves antioxidant status in ISP-induced cardiotoxicity in rats.

Pre-treatment with α-tocopherol and Terminalia arjuna ameliorates, pro-inflammatory cytokines, cardiac and apoptotic markers in myocardial infracted rats.

OBJECTIVE: This study was aimed to evaluate the cardioprotective potential of combination of T. arjuna and α-tocopherol in isoproterenol induced myocardial injury. METHODS: Wistar albino rats were pre-treated with hydroalcoholic extract of T. arjuna (HETA) and α-tocopherol (100 mg/kg b. w) daily for 30 days. Isoproterenol (ISP, 85 mg/kg b.w) was administered on 28th and 29th days at an interval of 24 hr. RESULTS: ISP treated rats showed significant increase in lipid peroxidation (MDA), cardiac markers (CK-MB, SGOT, Trop I and LDH), pro-inflammatory cytokine (IL-6, CRP, TNF-α) levels and apoptotic markers (Bcl-2/Bax) as compared to healthy group. Pre-treatment with HETA 100 mg/kg b. w, reduced the elevated levels of these markers and significant effect (p<0.05) were observed with the combination of HETA and α-tocopherol at a dose of 100 mg/kg b. w, which was further confirmed by histopathological studies. CONCLUSION: The present study concluded that the combination of α-tocopherol (100 mg/kg b. w) and hydroalcoholic extract of T. arjuna (100 mg/kg b. w) augments endogenous antioxidant compounds of rat heart and also prevents the myocardium from ISP-induced myocardial injury and it may have therapeutic and prophylactic value in the treatment of ischemic heart disease.

Association of carotid intima-media thickness with leptin and apoliprotein b/apoliprotein a-I ratio reveals imminent predictors of subclinical atherosclerosis in psoriasis patients.

Psoriasis patients are often susceptible to cardiovascular diseases (CVD), including atherosclerosis. Traditional markers (biochemical and inflammatory) and diagnostic tools could detect occlusive but not subclinical atherosclerosis. Carotid intima-media thickness (CIMT), has recently been recognised as a non invasive diagnostic tool for identification of premature atherosclerosis. Therefore we evaluated 80 psoriasis patients and 80 age sex matched healthy controls for serum leptin levels and apolipoprotein B/apolipoprotein A-I ratio (apoB/apoA-I ratio) in relation with CIMT of carotid artery. Carotid intima-media thickness and carotid plaques were simultaneously measured by carotid sonography. Serum concentration of leptin and apolipoprotein were measured using enzyme-linked immuno sorbent assay (ELISA) and nephelometry respectively. Raised CIMT correlated to age of onset of the disease, serum leptin and apoB/apoA-I ratio in psoriasis patients. Taking into account, values that were above the 75 percentile of the three markers (leptin, apoB/apoA-I ratio and CIMT) the odds ratio was 4.26 (2.06-8.80 CI). Leptin and apoB/apoA-I ratio showed significant cumulative association with CIMT. Results of predictive analysis supports measurement of CIMT along with estimation of serum leptin and apoB/apoA-I ratio for prediction of premature atherosclerosis in psoriasis patients.

Small dense LDL: risk factor for coronary artery disease (CAD) and its therapeutic modulation.

Pathogenesis of coronary artery disease (CAD) is multi-factorial and many risk factors are associated with development of CAD. LDL-C has been an important target for therapeutic interventions and has been extensively studied. But, various studies have indicated that estimation of LDL-C is not enough to assess the risk. Moreover, LDL particles vary in their content, density and size which have different physico-chemical properties. In this paper, the role of small dense (sd) LDL in risk assessment for CAD and its response to different therapeutic modalities available have been reviewed.

Antihyperlipidemic effect of active principle isolated from seed of Eugenia jambolana on alloxan-induced diabetic rabbits.

Diabetes is accompanied by lipid abnormalities, which contribute significantly to cardiovascular morbidity and mortality in diabetic patients. We previously demonstrated the potent antihyperglycemic activity of the active principle (fraction II from Sephadex LH 20 chromatography [LH II]) isolated from ethanolic seed extract of Eugenia jambolana in diabetic rabbits. In the present study, the efficacy of LH II was evaluated for its hypolipidemic activity in alloxan-induced mildly diabetic (MD) and severely diabetic (SD) rabbits. Phytochemical investigation of LH II by various structural spectra showed the presence of saturated fatty acid, Δ(5) lipid, and sterol. Oral administration of LH II (10 mg/kg of body weight) for 21 days resulted in improved glycemic control in both MD and SD rabbits. After treatment with LH II, serum total cholesterol, triglycerides, high-density lipoprotein cholesterol, and the total cholesterol/high-density lipoprotein cholesterol ratio were significantly improved. LH II also resulted in significant (P < .001) improvement in 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity and levels of total lipids and glycogen in both MD and SD rabbits. Thus, the present study demonstrates that LH II possesses potent hypolipidemic activity and efficacy in both MD and SD rabbits.

Comparative effect of Ocimum sanctum, Commiphora mukul, folic acid and ramipril on lipid peroxidation in experimentally-induced hyperlipidemia.

Treatment with C. mukul and O. sanctum, showed a significant decrease in cholesterol and triglyceride levels respectively. O. sanctum also significantly increased serum HDL-cholesterol compared to control. Serum MDA levels were significantly reduced in all the treated groups compared to control suggesting that each of the drugs under study were effective in their free radical scavenging action. Erythrocyte SOD activity was increased in all the treatment groups with C. mukul showing the maximum effect followed by O. sanctum, folic acid and ramipril. The erythrocyte CAT activity was significantly increased in all the drug treated groups with maximum increase seen in O. sanctum and ramipril treated groups, whereas lesser effects were observed with C. mukul and folic acid groups. Thus, the indigenous drugs, C. mukul and O. sanctum had beneficial effect on hypercholesterolemic rabbit model, both in terms of lipid profile as well as antioxidant potential. Ocimum sanctum was found to be the most promising of all the drugs. Moreover, it could be hypothesized that these plant products along with folic acid and ramipril can be explored for synergistic effect for treatment for hypercholesterolemic conditions.

Elucidation of mechanism of action of Cassia auriculata leaf extract for its antidiabetic activity in streptozotocin-induced diabetic rats.

Cassia auriculata traditionally has been used to treat diabetes from ancient times. The objective of the present study was to investigate the mechanism of action for the antidiabetic activity of aqueous leaf extract of C. auriculata (CLEt) in streptozotocin-induced mildly diabetic (MD) and severely diabetic (SD) rats. CLEt was orally administered to MD and SD rats at a dose of 400 mg/kg once a day for 15 days. CLEt-treated MD and SD rats showed significant reduction in fasting blood glucose. Assessment of plasma insulin and C-peptide following treatment with CLEt revealed significant elevation in their levels. Administration of CLEt enhanced the activity of hepatic hexokinase and phosphofructokinase and suppressed glucose-6-phosphatase and fructose-1,6-bisphosphatase in both MD and SD rats. A significant rise in glycogen content was also observed in both liver and muscles of CLEt-fed MD and SD rats. Histopathological examination of pancreatic sections revealed increased number of islets and beta-cells in CLEt-treated MD as well as SD rats. The findings of the study suggest that the antidiabetic effect of CLEt could be due to its insulinogenic action. In addition, impaired glucose homeostasis was improved by feeding the extract through amelioration in the carbohydrate metabolic pathways. Thus, the extract may exert an antidiabetic effect through pancreatic as well as extrapancreatic action.

Cardiovascular friendly natural products: a promising approach in the management of CVD.

Natural products play an important role as nutritional supplements and provide potential health benefits in cardiovascular diseases (CVD). Compiling data from experimental, epidemiological and clinical studies indicates that dietary nutrients have profound cardioprotective effects in the primary as well as secondary prevention of coronary heart disease, hence they are considered as cardiovascular friendly natural products. The mechanism of cardioprotection produced by dietary nutritional supplements such as flavonoids (citrus fruits, pulses, red wine, tea and cocoa), olive oil, omega-3 (omega-3) fatty acids (fish oil and fish-based products), lycopene (tomato and tomato-based products), resveratrol (grapes and red wine), coffee, and soy in the prevention and treatment of cardiovascular disorders have been discussed in the present review, with the emphasis of epidemiological and clinical studies. Based on the intriguing results of various studies, prophylactic and therapeutic potential of cardiovascular friendly natural products have been suggested. The supplementation of cardiovascular friendly natural products needs to be considered in all populations who have high prevalence of CVD.

Ameliorative effect of Cassia auriculata L. leaf extract on glycemic control and atherogenic lipid status in alloxan-induced diabetic rabbits.

Oral administration of aqueous leaf extract of Cassia auriculata L. (100, 200, 400 and 600 mg/kg body wt daily for 21 days) to alloxan-induced mild diabetic (MD) and severe diabetic (SD) rabbits produced dose dependent fall in fasting blood glucose up to 400 mg/kg dose from day 3 to day 21. Further, a significant elevation in the levels of insulin and reduction in glycosylated haemoglobin (HbA1c) was observed in both MD and SD rabbits when treated with 400 mg/kg dose of the extract. The significant decrease in serum levels of triglycerides (TG), total cholesterol (TC) and low-density lipoprotein-cholesterol (LDL-C) with a concomitant increase in high-density lipoprotein-cholesterol (HDL-C) was exhibited by MD as well as SD rabbits following treatment with the extract. Atherogenic indices (TG/HDL-C, TC/HDL-C and LDL-C/HDL-C) were also significantly reduced in both diabetic models of rabbits fed with the extract. Effect of the extract at 400 mg/kg dose was comparable to that of glibenclamide (600 microg/kg), a reference antidiabetic drug. Thus, the present study demonstrated that aqueous leaf extract of C. auriculata can be a possible candidate for antidiabetic drug.

hs-CRP and oxidative stress in young CAD patients: A pilot study.

Premature coronary artery disease (CAD) is common in India. We, therefore, studied oxidative stress, dyslipidemia, and high sensitivity-C reactive protein (hs-CRP) levels in young CAD patients. Present study consisted of male CAD patients below 40 years and age and sex matched healthy controls (n=30 each). Fasting blood samples were analyzed for serum lipid profile, malondialdehyde, antioxidant enzymes and hs-CRP levels. Dyslipidemia was observed in 90% of the young CAD patients, of which 72.2% showed increased serum triglycerides and decreased HDL-cholesterol. LDL-cholesterol levels were high in 77.8%. Serum malondialdehyde and hs-CRP levels were increased significantly (p<0.0001) as compared to controls. hs-CRP levels were in high risk range in all the young patients. However, glutathione peroxidase activity was reduced significantly (p<0.05). Our data suggests that elevated hs-CRP levels along with dyslipidemia and oxidative stress adds to the predictive value of premature CAD in young Indians.