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Home > Bioassay Guided Isolation of Secondary Metabolites for Antidiabetic and Anti-Amnesic Potential of Elaeagnus Umbellata Thunb

Bioassay Guided Isolation of Secondary Metabolites for Antidiabetic and Anti-Amnesic Potential of Elaeagnus Umbellata Thunb

Thesis Info

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Author

Nausheen, ,

Program

PhD

Institute

University of Malakand

City

Malakand

Province

KPK

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Chemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/12441/1/Nausheen%20chemistry%202019%20uom%20malakand%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725580669

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The creatures on the earth are in search of discovering new things since the start of life on earth. All these efforts are for making their lives better in all aspects of life. With the passage of time human being faced various diseases from minor to chronic and are still facing. In ancient civilization people used herbal products to treat human and animal diseases. Human are in search of discovering new potent and effective medicine from medicinal plants. All plants yield chemical compounds called primary metabolites required for their basic metabolic activities. Apart from primary metabolites they also produces secondary metabolites with more specific and are not used for energy productions but are used for attracting insects which help in pollination and some of them are even used for defence against invaders. They also have beneficial functions in humans if eaten/used as drugs. Such metabolites can be used to produce potent drugs for the modern era diseases. Due to the instinct nature of human from the day first he is behind in search of better in every aspect of life. Although 800 plants have been reported to have antidiabetic activities however none of them is perfect to resolve the issue completely. Due to the mentioned fact above we design this study in the hope that probably this plant; Elaeagnus umbellata (silver berry) will be more potent than the already reported plants. Our decision of selecting this plant as antidiabetic and antiamnesic is not random we have selected this plant due its phytochemical composition and inhibitory potential. To best of our knowledge the activities we carried have not been reported before. Although other berries fruits of the other families have already reported to have antidiabetic affects. Elaeagnus umbellata belongs to Elaeagnaceae family are one of the wild spiny branched shrubs widely distributed in Himalayan areas of Pakistan. E. umbellata barriers are sources of phenolic and other bioactive compounds that scavenge the reactive oxygen species (ROS) formed during metabolic processes. ROS has been the causative agent of various chronic ailments like diabetes mellitus, cancer, cardiovascular illness, arteriosclerosis, ischemic, nephritis, rheumatism, and neurodegenerative disorders like Alzheimer’s disease (AD). Elaeagnus umbellata fruit methanolic extract and their subsequent fractions were evaluated for curing of diabetes and neurological complaints. In the current study E. umbellata fruit berry have been evaluated for the mentioned diseases in form of crude methanolic extract, their subsequent fractions, essential oil and isolated pure compounds. The crude methanolic extract was produced by maceration process. The fractions were obtained by fractionation procedure based on solvent polarity beginning from lower to highly polar solvent (n-hexane, chloroform, ethyl-acetate, n-butanol and finally aqueous fraction was left yielding 95, 210, 115, 90 and 220 grams of extracts, respectively). HPLC-UV was used for the identification of phenolics present in the extract/fractions and their distribution amongst the fractions (HPLC finger printing). Essential oil were obtained through hydro distillation using Clevenger apparatus. The extracted components/fractions were subjected to column isolation to isolate pure compounds. Thin layer chromatography was used to confirm the isolation of a compound in pure state. The isolated compounds were purified via pencil/pen gravity column with small diameter. The compounds were finally identified through various spectroscopic techniques like; Furrier transform infra-red (FTIR), EI-MS, HNMR and C13NMR spectroscopic techniques while the essential oil was analysed through GC-MS for identification of phytoconstituents. E. umbellata berries methanolic extract/fractions, essential oil and isolated compounds were examined for in vitro and in vivo inhibition against α- amylase and α- glucosidase enzymes and anti-hyperglycemic effects in type 2 diabetes mellitus. The acetyl cholinesterase and butyryl cholinesterase inhibition potential and in vivo antiamnesic potential were also determined for extract/fractions and isolated compounds. Phytochemical analysis, antibacterial activity, total phenolic and flavonoid contents were determined for methanolic extract and subsequent fractions. On the basis of their antioxidant potential against DPPH (2, 20-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) free radicals, the Me-Ext, sub fractions (Chf-Ext, EtAc-Ext) and compound-I were found to be more active against α-amylase and α-glucosidase with IC50 values 400, 58, 200, 32 μg/ml and 200, 60, 140, 30 μg/ml respectively. Amongst different fractions of Elaeagnus umbellata fruit; the Chf-Ext and EtAc-Ext and compound-V showed maximum % inhibition against AChE (87±1.2, 84±1.0, 89±2.1) with IC50 values 33, 55, 31 μg/mL and BChE (89±0.3, 82±0.5, 90±2.1) at 1000 μg/mL concentration with IC50 values 35, 58, 30 μg/mL respectively. Study of acute toxicity for Met-Ext/fractions (100-2000 mg/kg) and compound-V (10, 15, 40, 50 and 100 mg/kg/b/w) of E. umbellata were carried out which did not produce any significant behavioral alterations (respiratory aches, convulsions shortage, writhing, variations to reflex actions or mortality) in adults Sprague Dawley rats weighing 150-200 g. An insignificant increase in petulance was detected at 2000 mg/kg dose for Met-Ext/fractions and 100 mg/kg dose for compound-V in three animals out of total eight. All animals appeared healthy at the end one solar day cycle i.e. 24 hour to 1 week with no noticeable variations in appearance or behavior. No mortality was noticed up to one week. All procedures related to animal were carried out according to the Animal Scientific Procedure Act; UK (1986). In vivo results indicates that the Met- Ext, Chf-Ext, EtAc-Ext fractions and compound-I were found to be more potent in curing the hyperglycemia in STZ- induced type 2 diabetes mellitus in rats as considerable reduction of glucose level was observed when compared to the standard glibenclamide drug (0.5 mg/kg, p.o.). Furthermore, considerable reduction in serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase, total cholesterol, low density lipoproteins and triglycerides were observed indicating useful effects of extracts on secondary complications associated with type 2 diabetes mellitus. In vivo antiamnesic activity was carried out to evaluate the short and long term memory using the behavioural trials; Y-Maze Spontaneous Alternation Behaviour and recognition test for novel object. In albino mice the Chf. Ext fraction and compound-I (CGA) overturned the amnesia prompted by scopolamine and significant increase in SAP for Chf. Ext (200 mg/kg) fraction and CGA (30 mg/Kg) compound analogous to donepezil (2 mg/kg) drug in the Y-maze procedure. Similarly, Chf. Ext (100 mg/Kg) and CGA (10 mg/Kg) shown significant (p < 0.05) increase in %DI related to scopolamine (1 mg/kg). Finally the molecular docking was applied to identify some common interactions observed between acarbose and all docked compounds in the active sites of both α-amylase and α-glucosidase enzymes that have shown the inhibitory effects of the mentioned enzymes. The mechanisms of binding were also analyzed on a GOLD suit v5.6.3 against AChE & BChE enzyme. The Analysis of binding modes indicated similar binding orientations for rutin, epigallocatechin gallate, chlorogenic acid, quercetin, catechin, morin, catechin hydrate, ellagic acid and donepezil in the active gorge of the receptor protein of AChE and BChE enzymes. In conclusion, E. umbellata fruit can be potentially recommended for controlling type 2 diabetes mellitus, oxidative stress caused by reactive oxygen species, memory impairments and neurological disorders. To search more potent antidiabetic and antiamnesic phytochemicals, more studies are required to formulate potent drugs and evaluate their proper mechanisms.
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