ANTIOXIDANT ACTIVITY AND FT-IR ANALYSIS OF DATURA INNOXIA AND DATURA METEL LEAF AND SEED METHANOLIC EXTRACTS.

BACKGROUND: Datura (family- Solanaceae), has a long history of being used as herbal medicine. These medicinal effects have been attributed to the phytochemicals present in the plant leaves and seeds, in particular alkaloids, flavonoids and phenolic compounds. The objective of this study was to investigate the methanolic leaf and seed extracts of Datura innoxia (DLP-I & DSP-I) and Datura metel (DLP-M & DSP-M) for their total phenolic, flavonoids and in-vitro antioxidant properties. MATERIALS AND METHODS: Determination of total phenolic content and total flavonoid content and antioxidant activity in terms of total antioxidant assay, ABTS assay, DPPH assay and in-vitro lipid peroxidation inhibiting activity were determined along with the FT-IR (Fourier transform infrared spectroscopy) analysis of the extracts. RESULTS: The highest total phenolic and total flavonoid content were registered by the D. innoxia leaf extract (70.26 ±1.12 mg GAE/g and 34.24 ± 1.28 mg RE/g respectively). Maximum DPPH radical scavenging activity was exerted by the leaf extract of D. innoxia (IC50 = 146.69 ± 8.46 µg/mL) among the four different methanolic extracts. The highest activity of the ABTS assay was found in Datura innoxia leaf extract (IC50 value = 149.42 ± 13.43 µg/mL) and the highest total antioxidant capacity was found to be present in D. innoxia leaf extract (221.25 ± 1.06 mg AAE/g) whereas D. metel seed extract registered the maximum lipid peroxidation inhibition activity (IC50 = 112 ± 1.30 µg/mL). The FT-IR data also supported the maximum activity in D. innoxia (leaf and seed) extracts. CONCLUSION: The results thus obtained suggested that the plant Datura innoxia possess considerable antioxidant activity over Datura metel and therefore can be established as a potential source of natural antioxidant.

Evaluation of antioxidant activity and total phenol in different varieties of Lantana camara leaves.

BACKGROUND: Phytochemicals like carotenoids, tocopherols, ascorbates and phenols present in the plants are strong antioxidants and have an important role in the health care system. There is growing interest in correlating the phytochemical constituents of a plant with its pharmacological activity. Therefore, the present study investigates the content of total phenolics, flavonoids and the antioxidant activity of four different varieties of Lantana camara L. (Verbenaceae) leaves by using in vitro antioxidant models. METHODS: The leaves of Chandigarh purple variety (CPV), Palampur red variety (PRV), Chandigarh yellow turning pink variety (YTPV) and Chandigarh yellow variety (CYV) Lantana camara were collected and the total phenolic, flavonoid content, antioxidant and free radical scavenging activities were determined in their methanolic extracts. RESULTS: The phenolic content was found to be highest in the CYV extract (232.99 ± 15.97 mg GAE/ g extract). The content of the flavonoids are in the order of YTPV, PRV, CPV and CYV. The IC50 values for the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging test were in the order of CYV (33.30 ± 2.39) < PRV (40.32 ± 2.94) < YTPV (475.33 ± 5.20) < CPV (927.16 ± 2.88 µg/mL). The highest total antioxidant capacity was observed in CYV (222.20 ± 5.05 mg AAE/ g). The Ferric ion reducing antioxidant potential (FRAP) value of the extracts were in the order of CYV > PRV > YTPV > CPV. The IC50 values of 2, 2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS) scavenging assay for CYV, PRV, YTPV, CPV were 18.25 ± 0.19, 18.24 ± 1.82, 50.43 ± 9.49, 52.84 ± 1.82 µg/mL respectively. PRV extract showed the maximum in vitro lipid peroxidation inhibition effect with an IC50 value of 68.50 µg/mL which is even stronger as compared to the standard Rutin (79.69 µg/mL). The extracts showed a strong correlation between the phenolic content and their antioxidant activities. The highest correlation (r = 0.998, R2 = 0.997) was found between total phenolic content and ABTS scavenging assay. CONCLUSION: Among the four varieties investigated, CYV and PRV extracts showed strong antioxidant activities and may be used as a potential source of natural antioxidant against free radical associated diseases.

Ethanol and age enhances fluoride toxicity through oxidative stress and mitochondrial dysfunctions in rat intestine.

Fluoride toxicity and alcohol abuse are the two serious public health problems in many parts of the world. The current study was an attempt to investigate the effect of alcohol administration and age on fluoride toxicity in rat intestine. Six and 18 months old female Sprague Dawley rats were exposed to sodium fluoride (NaF, 25 mg/kg), 30 % ethanol (EtOH, 1 ml/kg), and NaF+EtOH (25 mg/kg+1 ml/kg) for a period of 20, 40, and 90 days. The levels of lipid peroxidation were increased, while the content of reduced glutathione, total, and protein thiol was decreased with NaF treatment. Under these conditions, animals showed an age-related decline in the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase which were further aggravated upon NaF or/and EtOH treatment. Mitochondrial respiration rate and the activities of complexes I, II, and IV enzymes of electron transport chain were decreased, while the levels of nitric oxide and citrulline were increased with age and NaF or/and EtOH treatment. Histological examination revealed large reactive lymphoid follicles, excess of lymphocytes in lamina propria of villi, villous edema, focal ileitis, necrosis of villi, and ulceration in NaF- or/and EtOH-treated animals in both the age groups. These findings suggest that fluoride mediate its toxic effects on intestine through oxidative stress and mitochondrial dysfunctions which are further augmented with alcohol consumption and advancing age.

Effects of fluoride and ethanol administration on lipid peroxidation systems in rat brain.

Exposure to fluoride and excessive ethanol consumption has been identified as a serious public health problem in many parts of the world, including India. Thus, the effect of co-exposure to fluoride and ethanol for 3-6 weeks was studied on lipid peroxidation (LPO) and oxidative stress related parameters in the rat brain. After 3 weeks, co-treated animals showed 95% increase in LPO levels compared to control. However, the levels of reduced glutathione, total and protein thiols were decreased. These changes were accompanied by a decrease in the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase. Rats exposed to fluoride together with ethanol for 6 weeks resulted in 130% increase in LPO and decrease in the reduced glutathione levels. The activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase were reduced under these conditions. Brain histology revealed excessive lymphocytes, edema and spongeosis in the cortical region after six weeks of fluoride and ethanol treatment. These results suggest that exposure to fluoride together with ethanol enhances lipid peroxidation by affecting antioxidant defence systems in the rat brain.

Modulation of lipid peroxidation and antioxidant defense systems in rat intestine by subchronic fluoride and ethanol administration.

Excessive consumption of fluoride and ethanol has been identified as injurious to human health. Fluoride and ethanol co-exposures are commonly seen among the alcoholics residing in endemic fluoride areas worldwide. This study was undertaken to examine the modulation of lipid peroxidation and antioxidant defense systems in rat intestine by subchronic fluoride and ethanol administration. Female Sprague-Dawley rats were divided into four groups: group I (control), group II (fluoride was given orally at a dose of 25 mg/kg body weight), group III (30% ethanol was given orally at a dose of 1 mL/kg body weight), and group IV (a combination of fluoride and ethanol was administered orally at the dose described for groups II and III). Lipid peroxidation was elevated (P<.05) in intestine of rats by fluoride or ethanol treatments for 20 or 40 days. However, glutathione content was reduced by fluoride (32 and 44%) and ethanol (21 and 40%) treatments after 20 and 40 days, respectively. Fluoride-exposed animals showed reduction (P<.05) in the activities of superoxide dismutase (22 and 42%), catalase (30 and 37%), glutathione peroxidase (22 and 35%), glutathione reductase (32 and 34%), and glutathione-S-transferase (24 and 30%) after 20 and 40 days. A similar decrease (P<.05) in the activities of these enzymes was also noticed in animals exposed to ethanol for 20 or 40 days. The observed changes in lipid peroxidation, reduced glutathione levels, and enzyme systems were further augmented in intestine of rats exposed to fluoride and ethanol together. Intestinal histology showed large reactive lymphoid follicles along with mild excess of lymphocytes in lamina propria of villi, villous edema, focal ileitis, and necrosis of villi in animals exposed to fluoride and ethanol for 40 days. These findings suggest that fluoride and ethanol exposure induces considerable changes in lipid peroxidation, antioxidant defense, and morphology of rat intestine, which may affect its functions.

Evaluation of antimotility effect of Lantana camara L. var. acuelata constituents on neostigmine induced gastrointestinal transit in mice.

BACKGROUND: Lantana camara L. (Verbenaceae), a widely growing shrub which is toxic to some animal species, has been used in the traditional medicine for treating many ailments. The purpose of the present study was to evaluate the antimotility effects of Lantana camara leaf constituents in mice intestine. METHODS: Evaluation of antimotility activity was done in intestine of mice treated with Lantana camara leaf powder, Lantana camara methanolic extract (LCME), lantadene A, neostigmine and neostigmine + LCME. Neostigmine was used as a promotility agent. Intestinal motility was assessed by charcoal meal test and gastrointestinal transit rate was expressed as the percentage of the distance traversed by the charcoal divided by the total length of the small intestine. The antidiarrheal effect of LCME was studied against castor oil induced diarrhea model in mice. RESULTS: The intestinal transit with LCME at a dose of 500 mg/kg was 26.46% whereas the higher dose (1 g/kg) completely inhibited the transit of charcoal in normal mice. The % intestinal transit in the neostigmine pretreated groups was 24 and 11 at the same doses respectively. When the plant extracts at 125 and 250 mg/kg doses were administered intraperitonealy, there was significant reduction in fecal output compared with castor oil treated mice. At higher doses (500 and 1000 mg/kg), the fecal output was almost completely stopped. CONCLUSION: The remarkable antimotility effect of Lantana camara methanolic extract against neostigmine as promotility agent points towards an anticholinergic effect due to Lantana camara constituents and attests to its possible utility in secretory and functional diarrheas and other gastrointestinal disorders. This effect was further confirmed by significant inhibition of castor oil induced diarrhea in mice by various doses of LCME.

Ethanol-induced changes in lipid composition of intestinal microvillus membrane in rats fed different dietary fats.

BACKGROUND/METHOD: The effect of feeding ethanol for 5 weeks on the lipid composition of the intestinal microvillus membrane (MVM) was studied in rats fed a commercial rat pellet (RP) diet or purified diets containing 10% coconut oil (CCO), corn oil (CO) or fish oil (FO). RESULTS: A low cholesterol/phospholipid ratio and increased saturated fatty acid level were observed in MVM from the CCO or FO groups. Chronic administration of ethanol to RP- or CO-fed animals increased phospholipids, total and free cholesterol, and the triglyceride and ganglioside content of MVM. The free cholesterol and phospholipid content was reduced while the triglyceride level remained unaffected by ethanol treatment in the CCO or FO groups. Ethanol ingestion decreased 10:2 and 20:4 (n-6 fatty acids) but increased the saturated fatty acid content of MVM in all the dietary groups except in CCO-fed animals where the 18:2 level was not affected. An elevated 18:1, but decreased 22:6 percentage was observed in the ethanol-fed FO group. The fatty acid composition of MVM from the CCO-fed group was least affected by ethanol treatment. CONCLUSION: These observations suggest that the type of dietary fat modifies ethanol-mediated alterations in MVM lipid composition.