A simple method for extracting both active oily and water soluble extract (WSE) from Nigella sativa (L.) seeds using a single solvent system

The current study provides a way of extraction for both active NSO and WSE from Nigella sativa seeds using 98% methanol. About 1 kg of ground seeds was macerated by 1:2.5 w/v (g/mL) for 72 hours. After rotary evaporation and 7 days of continuous drying and chilling at 50 and 4 °C, NSO and WSE were obtained at the same instant. Solubility tests of 24 solvents and 11 thin layer chromatographic analyses while 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging assay of NSO (73.66) , WSE (33.32) and NSO + WSE (78.22) against ascorbic acid (IC50 = 4.28 mg/mL) was performed. WSE was found to be highly soluble in water and 5% NaOH exhibiting the same Rf value of 0.95 for EtOH:DMSO (9:1) against the honey. WSE has revealed more than twofold higher anti-oxidant activity than others. Formulation of WSE with Tualang honey may provide better targeted hydrophilic drug delivery systems

THE NIGELLA SATIVA (L.) SEEDS EXTRACT TREATMENT ON THE HIPPOCAMPAL GFAP IN 2VO-INDUCED CHRONIC NEUROINFLAMMATION

Neuroinflammation has been closely linked to neurodegeneration that leads to dysfunction of memory and learning where glial fibrillary acidic protein plays important role within hippocampus. In experimental neurodegeneration achieved by two-vessel occlusion (2VO) intervention, Nigella sativa (L.) seeds extract (NSSE) has shown to have anti-neuroinflammatory and antioxidant properties. Toxic free radicals formation and prevention of neuroinflammatory mediators are of two possible mechanisms. The current study aimed to investigate the anti-neuroinflammatory effect of Nigella sativa and the role of GFAP as a relevant biomarker of neuroinflammation. The relative normalized hippocampal GFAP mRNA expressions (∆∆Cq) of Nigella sativa (L.) seeds extract treatment (100mg/kg/day orally) was compared with that of healthy control (HC) and untreated 2VO groups (n=6) of rats. The brain hippocampal tissues were humanely collected at the end of 10th treatment week and preserved in Allprotect™ reagent at – 80°C. Total RNA was extracted and purified by phenol/chloroform method using kits, and reverse transcribed into cDNA and relatively quantified as per ∆∆Cq. The NSSE treatment showed significantly (P<0.001) different relative GFAP mRNA expression in the treated group as compared to that of untreated 2VO whilst it was insignificantly (P>0.5) different to that of HC. Prolonged or daily treatment with NSSE may possess moderate anti-neuroprotective activity within hippocampus.&nbsp

The thrombolytic and cytotoxic effects of nigella sativa (L.) seeds: the prophetic medicine

The Water-Soluble Extract (WSE) is a crude bioactive phytoconstituent of Nigella sativa (L.) seeds discovered recently. The current findings report about the thrombolytic and cytotoxic effects of WSE using human blood clot lysis and brine shrimp lethality (BSL) bioassay. The thrombolytic effect of WSE (1,666.67 µg/mL) was determined via the clot and lysate weight measurements compared to streptokinase (STK) of 30,000 IU/mL and normal saline (NS) while the cytotoxicity of WSE (44.14-2,000 µg/mL) against vincristine sulfate (VCS;3.125-100 µg/mL). WSE has shown extremely statistically significant (p<0.0001) clot lysis (90.00%) compared to NS (3.76%) whilst it was also significantly different (p<0.0063) to STK (72.41%) exhibiting LC50 of 1,795.90 µg/mL vs. VCS (39.25 µg/mL) in a dose-dependent manner. The current results suggested WSE has a potent thrombolytic effect with mild dose-dependent cytotoxicity towards brine shrimp nauplii (Artemia salina). It also suggested WSE might have enzymatic roles on thrombin, fibrin, and plasmin of blood. This pharmacological action of WSE is might be due to its antioxidant property, short-chain fatty acids and/or amino acids. Further studies are highly recommended on the enzymatic role(s) and bioactive phytoconstituents of WSE

Hippocampal GFAP and MAP2 mRNA expressions of curry leaves extract treatment in 2VO rats of neurodegeneration

Background: Experimental neurodegeneration can be induced by two-vessel occlusion intervention (2VO). Nigella sativa (Linn.) seeds extract (NSSE) has recently shown to enhance memory and learning task performances in rats with 2VO. Prevention of neuroinflammation and/or of toxic free radical formation are possible mechanisms. The glial fibrillary acidic protein (GFAP) is a vital neuroinflammatory marker in neuroscience researches. GFAP was found to be up-regulated in 2VO rats indicating astrogliosis. Microtubule associated protein type II (MAP2) is an important marker for neuroprotection during and after ischemic brain injury or hypoxia. It was found to be down-regulated in 2VO rats. Objective: The current study aimed to compare the hippocampal MAP-2 and GFAP mRNA expressions of 2VO (untreated) male rats in the experimental neurodegeneration with other groups of rats treated with NSSE. Method: 18 adult male Sprague Dawley rats were randomly divided into 3 groups (n=6); Healthy Control (HC); 2VO- without treatment (2VO); 2VO+NSSE-treated (NSSE). The NSSE group was pre-treated (2ml, 50 mg/kg/day, orally) for 10 days prior to 2VO surgery and continued until all animals were sacrificed at the end of 10th postoperative week. Hippocampal samples were then collected; total RNA was extracted, purified and relatively quantified via reverse transcribed cDNA as per ∆∆Cq of RT-qPCR assay. Results: There was a significant 3 folds difference of GFAP mRNA expression in both HC and NSSE groups as compared to that of 2VO. However, GFAP mRNA expression difference was not significant (P=0.52) of for NSSE vs. HC. Hippocampal MAP2 mRNA expression of NSSE group appeared to be insignificantly different from the untreated 2VO group. Conclusion: Prolonged treatment with NSSE has the potential to prevent hypoperfusion-induced neuroinflammation within the hippocampus. This may be a potential mechanism for neuroprotection against mild cerebral ischemic injuries. However, NSSE was unable to prevent loss of neuronal plasticity

Effects of nigella sativa (Linn.) seeds extract treatment on the hippocampal MAP2 and GFAP mRNA expressions in the experimental neurodegeneration

Background: Experimental neurodegeneration can be induced by two vessel occlusion intervention (2VO). Nigella sativa (Linn.) seeds extract (NSSE) has recently shown to enhance memory and learning task performances in rats with 2VO. Prevention of neuroinflammation and/or of toxic free radical formation are possible mechanisms. The glial fibrillary acidic protein (GFAP) is a vital neuroinflammatory marker in neuroscience researches. GFAP was found to be up-regulated in 2VO rats indicating astrogliosis. Microtubule associated protein type II (MAP2) is an important marker for neuroprotection during and after ischemic brain injury or hypoxia. It was found to be down-regulated in 2VO rats. Aim: The current study aimed to compare the hippocampal MAP-2 and GFAP mRNA expressions of 2VO (untreated) male rats in the experimental neurodegeneration with other groups of healthy control and NSSE treated rats. Method: 18 adult male Sprague Dawley rats were randomly divided into 3 groups (n = 6); Healthy Control (HC); 2VO- without treatment (2VO); 2VO + NSSE-treated (NSSE). The NSSE group was pre-treated (2 ml/kg/day, orally) for 10 days prior to 2VO surgery and continued until all animals were sacrificed at the end of 10th postoperative week. Hippocampal samples were then collected; total RNA was extracted, purified and relatively quantified via reverse transcribed cDNAasperΔΔCq of RT-qPCR assay. Results: There was a significant 3 folds difference of GFAP mRNA expression in both HC and NSSE groups as compared to that of 2VO. However, GFAP mRNA expression difference was not significant (P = 0.52) of for NSSE vs. HC. Hippocampal MAP2 mRNA expression of NSSE group appeared to be insignificantly different from the untreated 2VO group. Conclusion: Prolonged treatment with NSSE has the potential to prevent hypoperfusion-induced neuroinflammation within the hippocampus. This may be a potential mechanism for neuroprotection against mild cerebral ischemic injuries. However, NSSE was unable to prevent loss of neuronal plasticity