Oleander attenuates hepatic inflammation in a TLR4-independent manner and by favorable modulation of hepatocellular global metabolome that supports cytoprotection.

ETHNOPHARMACOLOGICAL RELEVANCE: Nerium oleander is used to treat liver-associated chronic metabolic diseases in traditional medicinal systems across the globe. The hepatoprotective effects of oleander are mentioned in Indian and Chinese traditional medicinal literature. AIM OF THE STUDY: The present study aimed to investigate the cellular mechanisms behind the hepatoprotective effects of a non-toxic dose of oleander (NO). MATERIALS AND METHODS: The hepatoprotective effects of NO were tested against lipopolysaccharide (LPS)-treated HepG2 cells. Oxidative stress response was studied using cellular enzymatic assays, and gene expression was analyzed using qRT-PCR. HepG2 cells were pretreated with TAK-242 (pharmacological inhibitor of TLR4) to decipher the anti-inflammatory mechanisms of NO. Cell-free metabolites were analyzed using GCMS and were subjected to pathway enrichment analysis. RESULTS: NO reduced systemic inflammation, serum lipid peroxidation byproducts, and glucose without affecting serum transaminase levels and hepatic histopathological features. NO attenuated the inflammation-induced loss of antioxidant enzyme activities and mRNA expressions of toll-like receptor-4 (TLR4)/nuclear factor κß (NFκß)-dependent inflammatory genes. In TAK-242 pretreated cells, LPS was unable to induce inflammatory and oxidative responses. However, NO treatment in TAK-242 pretreated cells with LPS stimulation further reduced the signs of inflammation and improved hepatoprotective activities. A comparative analysis of the intracellular global metabolome from HepG2 cells with and without NO treatment indicated NO-mediated favorable modulation of intracellular metabolic pathways that support cytoprotective activities. CONCLUSION: NO protects HepG2 cells from LPS-induced oxidative and inflammatory injury. The hepatoprotective effects of NO are mediated by a TLR4-independent process and through a favorable modulation of the intracellular global metabolome that supports cytoprotection.

Correlative metabolomic fingerprinting and molecular docking studies of dermatological phytotherapeutics of South-Eastern Himalaya.

Viburnum erubescens Wall., Rhododendron arboretum Sm., Eurya japonica Thumb., Symplocos lucida (Thunb.) Siebold & Zucc, and Symplocos pyrifolia Wall. ex G. Don are extensively used by the native and ethnic populations of the South-Eastern Himalayan region for several dermatological conditions, yet their phytochemical composition remained largely unknown. Therefore, the aim of the study was to explore the therapeutically relevant volatile phytochemical compositions and study the molecular interactions against intracellular cytoregulatory transcription factors. Leaves of the five plants were subjected to Gas chromatography-Mass spectrometry (GCMS) post silylation derivation. The results were further analyzed using multivariate statistical methods such as Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA). A total of 115 compounds were identified in the five plants. Multivariate analysis revealed optimum metabolomic correlation between S. pyrifolia and S. lucida (0.876), whereas lowest correlation was found between E. japonica and V. erubescens (-0.242). Arbutin, ß-amyrin, betulin, ß-sitosterol and stigmasterol demonstrated highest interaction with the molecular targets. Collectively, the present study revealed the bioactive volatile phytochemicals responsible the therapeutic uses against diverse skin conditions.

Amelioration of CCl4 induced liver injury in swiss albino mice by antioxidant rich leaf extract of Croton bonplandianus Baill.

The progress in industrialization has blessed mankind with a technologically superior lifestyle but poor management of industrial waste has in turn poisoned nature. One such chemical is carbon tetra chloride (CCl4), which is a potent environmental toxin emitted from chemical industries and its presence in the atmosphere is increasing at an alarming rate. Presence of CCl4 in human body is reported to cause liver damage through free radical mediated inflammatory processes. Kupffer cells present in the liver are potentially more sensitive to oxidative stress than hepatocytes. Kuffer cells produced tumor necrosis factor-α (TNF-α) in response to reactive oxygen species (ROS), that might further cause inflammation or apoptosis. In this study hepatoprotective capacity of antioxidant rich extract of Croton bonplandianus Baill. (CBL) was evaluated on CCl4 induced acute hepatotoxicity in murine model. Hydro-methanolic extract of C. bonplandianus leaf was used for evaluation of free radical scavenging activity. Liver cells of experimental mice were damaged using CCl4 and subsequently hepatoprotective potential of the plant extract was evaluated using series of in-vivo and in-vitro studies. In the hepatoprotective study, silymarin was used as a positive control. Antioxidant enzymes, pro-inflammatory markers, liver enzymatic and biochemical parameters were studied to evaluate hepatoprotective activity of Croton bonplandianus leaf extract. Free radical scavenging activity of CBL extract was also observed in WRL-68 cell line. The phytochemicals identified by GCMS analysis were scrutinized using in-silico molecular docking procedure. The results showed that CBL extract have potent free radical scavenging capacity. The biochemical parameters were over expressed due to CCl4 administration, which were significantly normalized by CBL extract treatment. This finding was also supported by histopathological evidences showing less hepatocellularnecrosis, inflammation and fibrosis in CBL and silymarin treated group, compared to CCl4 group. ROS generated due to H2O2 in WRL-68 cell line were normalize in the highest group (200 µg/ml) when compared with control and negative control (CCl4) group. After molecular docking analysis, it was observed that the compound α-amyrin present in the leaf extract of C. bonplandianus has better potentiality to protect hepatocellular damages than the standard drug Silymarin. The present study provided supportive evidence that CBL extract possesses potent hepatoprotective capacity by ameliorating haloalkane induced liver injury in the murine model. The antioxidant and anti-inflammatory activities also affirm the same. The synergistic effects of the phytochemicals present in CBL are to be credited for all the hepatoprotective activity claimed above.

Acacia nilotica leaf improves insulin resistance and hyperglycemia associated acute hepatic injury and nephrotoxicity by improving systemic antioxidant status in diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Acacia nilotica (L.) Delile is used as a traditional anti-diabetic remedy in Bangladesh, Pakistan, Egypt, Nigeria and is mentioned in Ayurveda as well. AIM: The objective of the study was to evaluate the ethnomedicinal claim of A. nilotica leaf (ANL) extract for its efficiency in ameliorating diabetic complications. MATERIALS AND METHODS: ANL was orally administrated (50 and 200mg/kg) to alloxanized mice (blood glucose > 200mg/dL) for 20d. Parameters of glucose metabolism, hepatotoxicity, hyperlipidemia and nephrotoxicity were measured with emphasis on elevated oxidative stress. ANL was chemically characterized using GC-MS. Further, docking studies were employed to predict molecular interactions. RESULTS: ANL lowered (65%, P< 0.001) systemic glucose load in diabetic mice, which was otherwise 398% higher than control. ANL lowered (35%) insulin resistance, without any significant effect on insulin sensitivity (P> 0.05). Anti-hyperglycemic properties of ANL was further supported by lowering of HbA1c (34%; P< 0.001) and improved glucose utilization (OGTT). Overall diabetic complications were mitigated as reflected by lowered hepatic (ALT, AST) and renal (creatinine, BUN) injury markers and normalization of dyslipidemia. Elevated systemic oxidative stress was lowered by increased catalase and peroxidase activities in liver, kidney and skeletal muscle, resulting in 32% decrease of serum MDA levels. Apart from high phenolic and flavonoid content, tocopherol, catechol and ß-sitosterol, identified in ANL, demonstrated substantial binding affinity with Nrf2 protein (5FNQ) reflecting possible crosstalk with intracellular antioxidant defense pathways. CONCLUSION: The present study revealed the potentials of A. nilotica to alleviate diabetes-related systemic complications by limiting oxidative stress which justified the ethnopharmacological antidiabetic claim.

Variation in Phytochemical Composition Reveals Distinct Divergence of Aloe vera (L.) Burm.f. From Other Aloe Species: Rationale Behind Selective Preference of Aloe vera in Nutritional and Therapeutic Use.

In the present study, we have phytochemically characterized 5 different abundant Aloe species, including Aloe vera (L.) Burm.f., using silylation followed by Gas Chromatography-Mass Spectrometry technique and compared the data using multivariate statistical analysis. The results demonstrated clear distinction of the overall phytochemical profile of A vera, highlighted by its divergent spatial arrangement in the component plot. Lowest correlation of the phytochemical profiles were found between A vera and A aristata Haw. (-0.626), whereas highest correlation resided between A aristata and A aspera Haw. (0.899). Among the individual phytochemicals, palmitic acid was identified in highest abundance cumulatively, and carboxylic acids were the most predominant phytochemical species in all the Aloe species. Compared to A vera, linear correlation analysis revealed highest and lowest correlation with A aspera ( R2 = 0.9162) and A aristata ( R2 = 0.6745), respectively. Therefore, A vera demonstrated distinct spatial allocation, reflecting its greater phytochemical variability.

Radical Scavenging Activities of Lagerstroemia speciosa (L.) Pers. Petal Extracts and its hepato-protection in CCl4-intoxicated mice.

BACKGROUND: Lagerstroemia speciosa (L.) Pers. has medicinal importance. Bioactive phytochemicals isolated from different parts of L. speciosa, have revealed hypoglycemic, antibacterial, anti-inflammatory, antioxidant and hepato protective properties. Despite one report from Philippines detailing the use of L. speciosa as curative for fever and as well as diuretic, there is no experimental evidence about the hepatoprotective activity of the flower extracts. METHODS: Several spectroscopic methods, including GC-MS, were used to characterize phytochemicals present in the petal extract of L. speciosa. Ethanol extract of petals was evaluated for anti-oxidant and free radical scavenging properties by using methods related to hydrogen atom transfer, single electron transfer, reducing power, and metal chelation. This study has also revealed the in vitro antioxidant and in vivo hepatoprotective properties of petal extract against carbon tetra chloride (CCl4)-induced liver toxicity in Swiss albino mice. Hepatoprotection in CCl4 -intoxicated mice was studied with the aid of histology and different enzymatic and non-enzymatic markers of liver damage. Cytotoxicity tests were done using murein spleenocytes and cancareous cell lines, MCF7 and HepG2. RESULT: GCMS of the extract has revealed the presence of several potential antioxidant compounds, of them γ-Sitosterol and 1,2,3-Benzenetriol (Pyrogallol) were the predominant ones. The antioxidants activities of the flower-extract were significantly higher than curcumin (in terms of Nitric oxide scavenging activity; p = 0.0028) or ascorbic acid (in terms of 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) assay; p = 0.0022). The damage control by the flower extract can be attributed to the reduction in lipid peroxidation and restoration of catalase activity. In vitro cytotoxicity tests have shown that the flower extract did not affect growth and survivability of the cell lines. It left beyond doubt that a flower of L. speciosa is a reservoir of antioxidant and hepatoprotective agents capable of reversing the damage inflicted by CCl4-intoxication. CONCLUSION: Results from the present study may be used in developing a potential hepato-protective health drink enriched with antioxidants from Lagerstroemia speciosa (L.) Pers.

Toxicological assessment of Diplazium esculentum on the reproductive functions of male Swiss albino mouse.

CONTEXT: Diplazium esculentum, a commonly consumed seasonal vegetable, has been reported to have some pathological effects in some animals. But, its effect on the male reproductive function has not yet been studied. OBJECTIVE: To investigate the effects of boiled D. esculentum (BDE), the form which human consumes, on male reproductive functions of Swiss albino mice. MATERIALS AND METHODS: Male (120 in no.) and female (80 in no.) Swiss albino mice (6-8 weeks of age) were fed orally with 80, 160 and 320 mg/kg bw of BDE within a span of 180 d. After the treatment, body weight, absolute- and relative-testis weight, relative-weight of other organs, their biochemical parameters, hypo-osmotic swelling test (HOST) of spermatozoa, testis histology and fertility and fecundity tests were performed to justify the toxic effects of D. esculentum on male reproductive functions. RESULTS: Significant dose- and time-dependent decreases were observed in body weight, absolute- and relative-testis weight, relative-weights of other organs and their biochemical parameters, percentage of live spermatozoa and percentage of fertility and fecundity in BDE fed mice. Significant decreases were observed in diameter, perimeter and area of the seminiferous tubules of mice treated for 180 d. The percentage of empty seminiferous tubules was increased significantly in BDE treated mice when compared to the controls. DISCUSSION AND CONCLUSION: These results suggest that the intake of D. esculentum, even after cooking, may induce infertility by altering the male reproductive function, and therefore, should be evaluated further as a potential antifertility agent.

Haloalkane induced hepatic insult in murine model: amelioration by Oleander through antioxidant and anti-inflammatory activities, an in vitro and in vivo study.

BACKGROUND: Nerium oleander L. (syn. Nerium indicum Mill, Nerium odorum Aiton) belongs to the family Apocynaceae. It is used for its anti-inflammatory, anti-diabetic, anti-cancer and hepatoprotective activities in traditional medicine. Previous pharmacognostic studies suggested that 70 % hydro-methanolic extracts of oleander possess potent free radical scavenging and anti-inflammatory activities, both of which are helpful against hepatotoxicity. METHODS: Hydro-methanolic extracts of oleander stem and root were evaluated for their hepatoprotective activities in acute CCl4 intoxicated mouse through in vitro and in vivo studies. Silymarin was used as positive reference. Antioxidant enzymes, pro-inflammatory markers and liver enzymatic and biochemical parameters were studied. The extracts were further chemically characterized using Fourier Transform Infrared (FTIR) spectroscopy and Gas chromatography-mass spectrometry (GC-MS). RESULTS: CCl4 toxicity caused fatty liver formation by increase of relative liver weight (32.53 g) compared to control group (16.08 g). The elevated liver enzymatic and biochemical parameters due to CCl4 toxicity were considerably normalized by the extracts treatment under both in vivo and in vitro models. Oleander stem (NOSE) and root (NORE) extracts increased the reduced hepatic catalase activity 27.37 and 25.25 %, whereas peroxidase activity was increased 18.19 and 22.78 %, respectively. The extent of lipid peroxidation was significantly (p < 0.01) lowered 20.76 % (NOSE) and 21.12 % (NORE) compared to CCl4 group. The levels of pro-inflammatory tumor necrosis factor-α (TNF-α) was lowered 71.33 % (NOSE) and 61.60 % (NORE). Histopathological study demonstrated substantial reduction of hepatocellular necrosis, fatty infiltration, sinusoidal dilation, bile duct proliferation, vascular congestion, leukocyte infiltration in the silymarin and extract treated groups. Furthermore, various bioactive compounds were identified in the extracts such as apocynin, tocopherol, squalene, vanillin, isoeugenol, amyrin, lupeol etc. CONCLUSION: The present study provided convincing evidence that oleander extracts possess potent hepatoprotective capacity which was primarily governed by its antioxidant and anti-inflammatory activities. The collegial bioactivities of the phytochemicals may be accredited behind the hepatoprotective activity of oleander.

Comparative phytochemical profiling of Clerodendrum infortunatum with some selected medicinal plants predominant in the Sub-Himalayan region of West Bengal.

BACKGROUND: The aim of the present study was to quantify the phytochemicals of Clerodendrum infortunatum root and correlate on the basis of phytochemicals present with other medicinal plants belonging to different families. METHODS: Various standard biochemical and spectrophotometric methods were employed to study the phytochemical status of the root of C. infortunatum. All the analysis was performed in multiple sets. Various statistical methods like principal component analysis, dendogram, Euclidean distance and correlation study were done to compare the phytochemical profile of C. infortunatum with different selected medicinal plants belonging to different families. RESULTS: Results indicated the presence of various phytochemicals like phenol, alkaloid, flavonoid, tannin, thiamine, riboflavin, etc. Among the estimated phytochemicals, phenol had the highest quantity (65.59±1.26 mg/g) followed by lipid (27.51±0.23 mg/g) and tannin (16.40±0.07 mg/100 g). The phytochemical contents of C. infortunatum are much higher than those of the other reference plants. CONCLUSIONS: It may be concluded from the present study that the root of C. infortunatum contains very high amount of phytochemicals which are chiefly responsible for various medicinal properties. Therefore, the presence of the bioactive compounds in high quantity may lead to the potent medicinal value of C. infortunatum root.

Evaluation of anti-inflammatory activity and standardisation of hydro-methanol extract of underground tuber of Dioscorea alata.

Context The underground edible tuber of Dioscorea alata L. (Dioscoreaceae) is a functional food with high nutritive value and therapeutic potential. The tuber is known to possess anti-inflammatory properties in traditional medicine. Objective The present study explores the anti-inflammatory activity and standardisation of D. alata tuber hydromethanol extract. Materials and methods Hydromethanol extract (70%) of D. alata tuber was chemically characterised using HPLC and GC-MS techniques. Murine lymphocytes were cultured for 48 h with six different concentrations (0-80 µg/mL) of the extract. The expression of nitric oxide (NO), TNF-α, COX-1, COX-2, and PGE2 were evaluated using colorimetric and ELISA methods. Results Dioscorea alata extract inhibited the expression of NO and TNF-α with an IC50 value of 134.51 ± 6.75 and 113.30 ± 7.44 µg/mL, respectively. The IC50 values for inhibition of total COX, COX-1, COX-2 activities and PGE2 level were 41.96 ± 3.07, 141.41 ± 8.99, 32.50 ± 1.69, and 186.34 ± 15.36 µg/mL, respectively. Inhibition of PGE2 level and COX-2 activity was positively correlated (R(2) = 0.9393). Gallic acid (GA), 4-hydroxy benzoic acid (4HBA), syringic acid (SYA), p-coumaric acid (PCA), and myricetin (MY) were identified and quantified using HPLC. GC-MS analysis revealed the presence of 13 different phytocompounds such as hexadecanoic acid, methyl stearate, cinnamyl cinnamate, and squalene. Conclusion The D. alata extract significantly down-regulated the pro-inflammatory signals in a gradual manner compared with control (0 µg/mL). Different bioactive phytocompounds individually possessing anti-inflammatory activities contributed to the overall bioactivity of the D. alata tuber extract.

Immunomodulatory activity of Nerium indicum through inhibition of nitric oxide and cyclooxygenase activity and modulation of TH1/TH2 cytokine balance in murine splenic lymphocytes.

Nerium indicum is a medicinal plant which is used in the treatment of wide range of illness in different ethnopharmacological practices. We have studied the immunomodulatory activity of the 70 % hydro-methanolic extract of N. indicum leaves (NILE) by studying plaque forming cell assay, haemagglutination titre, cell proliferation assay, inhibition of nitric oxide expression and cyclooxygenase activity, estimation of IL-2, IFN-γ, IL-4, IL-10, TNF-α and IgM levels. Furthermore, we have characterized NILE with Fourier Transform Infra-red (FTIR) spectroscopy. The results displayed that NILE possessed potent immunomodulatory activity by up-regulating IL-2, IFN-γ, IL-10 expression and down-regulating IL-4, TNF-α expression in vitro. NILE also stimulated the in vivo expression of immunoglobulin level in mouse. FTIR analysis revealed phytocompounds of diverse chemical nature present in NILE. Thus, the potent immunomodulatory activity of N. indicum may have serious implications in the treatment of inflammatory diseases in the future.

Anti-inflammatory activity of Nerium indicum by inhibition of prostaglandin E2 in murine splenic lymphocytes.

OBJECTIVE: Nerium indicum Mill (syn. N. oleander L. and N. odorum Aiton; family: Apocynaceae) is a medicinal plant, used in the treatment of diverse ailments including various chronic inflammatory diseases in traditional medicine. We have previously demonstrated the immunomodulatory activity of a bioactive fraction of Nerium indicum leaf (NILE) by studying up-regulation of interleukin-2 (IL-2), IL-10, interferon-gamma and down regulation of IL-4, tumor necrosis factor-alpha (TNF-α), nitric oxide, cyclooxygenase-1 (COX-1) and COX-2 activities. Therefore, this study aimed to confirm the anti-inflammatory activity of NILE by inhibition of prostaglandin E2 (PGE2) activity in murine splenic lymphocytes in vitro. MATERIALS AND METHODS: Murine lymphocytes were isolated from spleen and stimulated with 5 ΅g/mL concanavalin A in RPMI-1640, supplemented with 50 U/mL penicillin, 50 U/mL streptomycin, 50 U/mL nystatin and 10% fetal bovine serum. Different concentrations (0-80 µg/mL) of NILE were added and the cells were cultured for 48 h. The culture supernatants were thereafter collected by centrifugation and assayed for expression of PGE2 level. The data were analyzed statistically. RESULTS: The results demonstrated a 2.26-fold inhibition of PGE2 level at 80 µg/mL of NILE. Half maximum inhibitory concentration (IC50) was calculated to be 44.95 ± 0.45 ΅g/mL. Linear correlation analysis of the dose-dependent PGE2 inhibition with other pro- and anti-inflammatory mediators demonstrated high inter-correlation between the parameters. CONCLUSIONS: Thus, the present study remains in accordance with our previous report and confirms the anti-inflammatory claim of N. indicum, mentioned in the traditional medicine.

Chemical characterization and assessment of antioxidant potentiality of Streptocaulon sylvestre Wight, an endangered plant of sub-Himalayan plains of West Bengal and Sikkim.

BACKGROUND: S. sylvestre Wright is an extremely rare plant, found only in the sub-Himalayan Terai region of West Bengal and neighboring Sikkim foot-hills. The plant has never been evaluated for any pharmaceutical properties. The phytochemical status of the plant is still unknown. Therefore, the aim of the study was to explore the antioxidant and free radical scavenging activities and analysis of bioactive compounds present in S. sylvestre. METHODS: S. sylvestre methanolic extract (SSME) was evaluated for different free radical scavenging activities such as hydroxyl radical, nitric oxide, singlet oxygen, hypochlorous acid, peroxynitrite, superoxide radical and hydrogen peroxide scavenging etc. Iron chelating capacity and inhibition of lipid peroxidation were studied in addition to the assessment of haemolytic activity and erythrocyte membrane stabilizing activity (EMSA). Chemical characterization of SSME were performed by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). RESULTS: The results indicate that SSME possess potent antioxidant activity with IC50 value of 113.06 ± 5.67 µg/ml, 63.93 ± 4.16 µg/ml and 142.14 ± 6.13 µg/ml for hydroxyl radical, superoxide radical and hypochlorous acid, respectively. HPLC analysis revealed presence of different phenolic secondary metabolites such as gallic acid, ferulic acid, p-coumaric acid, syringic acid, myricetin, quercetin etc. GC-MS analysis displayed the predominance of γ-sitosterol, vitamin E and squalene in SSME. CONCLUSION: The present study provides a convincing evidence that S. sylvestre not only possess potent antioxidant activity but also can be used as a source of natural bioactive phytochemicals in the future.

Assessment of hepatoprotective potential of N. indicum leaf on haloalkane xenobiotic induced hepatic injury in Swiss albino mice.

CCl4 is a potent environmental toxin which cause liver damage through free radical mediated inflammatory processes. In this study, hepatoprotective capacity of Nerium indicum leaf extract (NILE) was evaluated on CCl4 induced acute hepatotoxicity in murine model. Animals were divided into 5 groups and treated as following: control group (received only normal saline), CCl4 group (received only CCl4), silymarin group (received CCl4 and 100mg/kg silymarin), NILE low group (received CCl4 and 50mg/kg NILE) and NILE high group (received CCl4 and 200mg/kg NILE). After 10 consecutive days of treatment, the levels of hepatic biochemical markers, malondialdehyde (MDA) content, peroxidase and catalase activities were measured as well as histopathological study was performed. Furthermore, liver explant cultures were set up as following: control (no treatment), CCl4 group (contained 25 µl/ml CCl4), silymarin group (contained 25 µl/ml CCl4 and 100 µg/ml silymarin), NILE low group (contained 25 µl/ml CCl4 and 25 µg/ml NILE) and NILE high group (contained 25 µl/ml CCl4 and 100 µg/ml NILE). Hepatic transaminases and phosphatases, tumor necrosis factor-α (TNF-α) expression, nitric oxide (NO) release and cell viability were studied on the explant cultures. Phytochemical fingerprinting of NILE was performed by gas chromatography-mass spectrometry (GC-MS). The results showed that the biochemical parameters were overexpressed due to CCl4 administration, which were significantly normalized by NILE treatment. The findings were further supported by histopathological evidences showing less hepatocellular necrosis, inflammation and fibrosis in NILE and silymarin treated groups, compared to CCl4 group. GC-MS analysis revealed presence of different bioactive phytochemicals with hepatoprotective and antioxidant properties. Therefore, the present study indicate that NILE possesses potent hepatoprotective capacity to ameliorate haloalkane xenobiotic induced injured liver in murine model.

In vitro assessment of anticholinesterase and NADH oxidase inhibitory activities of an edible fern, Diplazium esculentum.

BACKGROUND: Diplazium esculentum is the most commonly consumed edible fern throughout Asia and Oceania. Several studies have been performed so far to determine different functional properties of this plant, but there have been no reports on the anticholinesterase and nicotinamide adenine dinucleotide (NADH) oxidase inhibitory activities of this plant. Therefore, the present study was conducted to determine the anticholinesterase and NADH oxidase inhibitory activities of 70% methanolic extract of D. esculentum. METHODS: The D. esculentum extract was investigated for its acetylcholinesterase and NADH oxidase inhibitory activities as well as its free radical scavenging and total antioxidant activities in the linoleic acid system. The free radical scavenging activity of the extract was determined by the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) method. The total antioxidant activity of the extract was evaluated by ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. RESULTS: The D. esculentum extract inhibited acetylcholinesterase and NADH oxidase in a dose-dependent manner, with IC50 values of 272.97±19.38 and 265.81±21.20 µg/mL, respectively. The extract also showed a potent DPPH radical scavenging activity with an IC50 value of 402.88±12.70 µg/mL. Moreover, the extract showed 27.41% and 33.22% of total antioxidant activities determined by FTC and TBA methods, respectively. CONCLUSIONS: Results indicated that 70% methanolic extract of D. esculentum effectively inhibited the enzymes acetylcholinesterase and NADH oxidase and acted as a potent antioxidant and free radical scavenger. These in vitro assays indicate that this plant extract is a significant source of natural antioxidants, which may be helpful in preventing the progression of various neurodegenerative disorders associated with oxidative stress.

Assessment of anti-diabetic activity of an ethnopharmacological plant Nerium oleander through alloxan induced diabetes in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Nerium oleander L. (syn. Nerium indicum Mill. and Nerium odorum Aiton.) is used for its anti-diabetic properties in Pakistan, Algeria, Morocco and is also recognized in Ayurveda. The present study was undertaken to investigate the anti-diabetic capacity of a standardized hydromethanolic extract of Nerium oleander in alloxan induced diabetes in mice. MATERIALS AND METHODS: Nerium oleander leaf extract (NOLE) was orally administered at 50 and 200mg/kg body weight (BW) dose to alloxanized mice (blood glucose >200mg/dl). After 20 consecutive days of treatment, various diabetic parameters were studied and compared with untreated mice. Furthermore, gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC) analysis was employed to reveal the phytochemical composition of the plant extract. RESULTS: NOLE demonstrated antihyperglycaemic activity by reducing 73.79% blood glucose level after 20 days of treatment. Oral glucose tolerance test (OGTT) revealed increase in glucose tolerance as evident by 65.72% decrease in blood glucose in 3h post treatment. Percentage decrease in different liver marker enzymes were significant along with decrease in triglyceride and cholesterol levels, displaying potent antihyperlipidemic activity. Peroxidase and catalase activity in liver, kidney and skeletal muscle were significantly restored besides marked reduction in lipid peroxidation and normalization of hepatic glycogen level in the NOLE treated alloxanized mice. Different bioactive phytocompounds with potent anti-diabetic activity were identified by GC-MS and HPLC analysis. CONCLUSION: The present investigation revealed that Nerium oleander possess potent anti-diabetic activity as claimed in different ethnopharmacological practices.

Pharmacological aspects of Nerium indicum Mill: A comprehensive review.

Phytomedicine is the oldest medical practice known to man. Since the dawn of mankind, various plant resources are used to cure different diseases and also for a long and healthy life. The ancient knowledge of plant based medicine has transferred from generations to generations and accumulated as ethnopharmacological knowledge among different ethnic groups. India is the spanning bed of traditional phytomedicinal system where Ayurveda was born out of the knowledge of traditional medicine. In various other countries of South-Eastern Asia, South America, and in Arabian countries, still today, a great number of people rely primarily on phytomedicines to cure diseases. In the complementary and alternative medicinal systems, Nerium indicum is one such plant which is famed for its therapeutic efficiency in different diseases globally. In the present time, when the pharmaceutical companies are concentrating more toward the plant based traditional medicines to avoid the side-effects and resistance against synthetic drugs, N. indicum has proved its efficiency in different disease models. Therefore, this review comprehensively covers the medicinal and pharmacological activities of different parts of the plant N. indicum.

Assessment of the immunosuppressive and hemolytic activities of an edible fern, Diplazium esculentum.

CONTEXT: Diplazium esculentum is the most commonly consumed fern throughout Asia and Oceania. Systemic toxicity and pathological effects on its consumption have already been demonstrated. But, the immunosuppressive and hemolytic activities of the boiled Diplazium esculentum (BDE), the form in which human consumes it, have not yet been studied. OBJECTIVE: To investigate the immunosuppressive as well as hemolytic activities, if any, of BDE in Swiss albino mice. MATERIALS AND METHODS: Body weight, relative spleen weight, plaque forming cell assay, hemaggutination antibody (HA) titer assay and macrophage counting were performed in BDE treated mice and respective control groups within a span of 180 days, and in vitro assays such as counting of cultured splenocytes, splenocytes proliferation assay and hemolytic assay were performed to justify the immunomodulatory as well as hemolytic activities of D. esculentum. RESULTS: Body weight and relative spleen weight were significantly decreased in BDE fed mice. Significant decreases were observed in the number of plaques formed, HA titer value and in the number of peritoneal macrophages within a span of 180 d. Significant dose-dependent decrease was observed in the number of cultured splenocytes. Significant dose-dependent increases in the percentage inhibition of splenocyte proliferation as well as percentage of hemolysis were evident by in vitro assays. DISCUSSION: These results suggest that the intake of D. esculentum may evoke immune dysfunction as well as may cause destruction of erythrocytes even after cooking. CONCLUSION: Therefore, the consumption of D. esculentum is alarming and may act as immunosuppressive agent.