Acta Scientific NUTRITIONAL HEALTH (ISSN:2582-1423) Volume 5 Issue 7 July 2021

Research Article Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

Mohammed A Hussein1*, Nada M El-Laban1, Lamiaa H Elabody1, Received: April 22, 2021 Amany M Ghazal1, Basant S Mohamed1, Azhar A Mohamed1, Lamiaa Y June 11, 2021 Elhalafwy1, Sanaa M Elsayad1, Amal S Othman2 and Ali A Ali3 Published: © All rights are reserved by Mohammed A 1Department of Biochemistry, Faculty of Applied Medical Science, October 6th Hussein., et al. University, Egypt 2Department of Med Labs, Faculty of Applied Medical Science, October 6th University, Egypt 3Vice President of Post Graduate Studies, October 6 University, Egypt

*Corresponding Author: Mohammed A Hussein, Professor of Biochemistry and

Medical Sciences, October 6 University, Egypt. Vice Dean for Post-graduate Studies and Scientific Research, Faculty of Applied

Abstract Neurodegenerative disorder clinically characterized by progressive cognitive and memory dysfunction. This study investigated the effect of Jasonia montana ethanolic extract (JMEE) on Ca‑, Mg+2 and Na+, K+-ATPase and acetylcholinesterase (AChE) activities

as well as β amyloid1-42 level in brain hippocampus of adult rats exposed to SCO. Rats were exposed to SCO (3.0 mg/Kg) and JMEE (150 mg/Kg) by gavage for 14 days. Rats were randomly divided into six groups with 6 rats in each: [Saline], [saline/SCO (3.0 mg/ kg)], [saline/SCO (3.0 mg/kg)/JMEE (150 mg/kg,)], [saline/SCO (3.0 mg/kg)/ DHC (3 mg/kg)] and [saline/SCO (3.0 mg/kg)/JMEE (150 mg/kg,)/ DHC (3 mg/kg)]. Results demonstrated that plasma TC, TG as well as brain hippocampus levels of AChE, MAO, βA1-42, +2, Mg+2 and Na+, K+ - TBARS. Also, the results showed that a significant depletion of plasma HDL-C as well as brain hippocampus levels of phospholipids, +2, Mg+2 GSH, ACh, Ca -ATPase. Treatment with JMEE (150 mg/Kg) prevented the increase in TC, TG, AChE, MAO, β amy and Na+, K+ loid1-42, TBARS activity when compared to SCO-treated group. JMEE treatment prevented the SCO-induced decrease in Ca Our data showed that JMEE have a protector effect against SCO induced neurodegenerative. Also, Jasonia Montana is a promising -ATPase activities as well as GSH, HDL-C, ACh and phospholipids in SCO-treated group when compared to normal group. therapeutic agent to attenuate neurological disorders associated with SCO induced dementia.

Keywords: Jasonia Montana; Antioxidant; Scopolamine; Acetylcholine Esterase; Monoamine Oxidase and Dementia

Abbreviations - - β amyloid1-42; TBARS: Thiobarbaturic Acid Reactive Substances; JMEE: Jasonia Montana Ethanolic Extract; DHC: Donepezil Hydro tathione. HDL-C: Cholesterol-High Density Lipoprotein; GHS: Reduced Glu chloride; AChE: Acetylcholinesterase; SCO: Scopolamine; TC: Total Cholesterol; TG: Triglycerides; MAO: Monoamine Oxidase; βA1-42:

Citation: Mohammed A Hussein., et al. “Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia". Acta Scientific Nutritional Health

5.7 (2021): 27-37. Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

Introduction its antioxidant , hypoglycemic , anticholestatic and28 anti-obesity activities. But there are no reports of the neuro- Neurodegenerative diseases are an incurable group of degen- [30] [31] [32] protective effect of Jasonia Monta. The present study was under- erative disorders. They are characterized by loss of neuronal cells, [33] leading to progressive impairments of functions of central nervous taken to investigate the investigated the effect of Jasonia Montana system (CNS) . Neurons are affected when a genetic mutation is extract antioxidant activity against scopolamine induced neurotox- - icity. [1] tations in amyloid precursor protein (APP) or presenilin 1 (PSEN1) identified in a ubiquitously expressed gene. For examples, the mu Materials and Methods and 2 (PSEN2) that occur in Alzheimer's disease (AD) , which Chemicals affect learning and memory circuits . [1] Scopolamine hydrobromide and donepezil hydrochloride was However, during aging and various[2,3] disease states, antioxidant defense systems can be altered leading to progressive oxidative obtained from Sigma Chemical Co. (St. Louis MO, USA). - Plant material tion . The brain is particularly sensitive to oxidative stress damage and subsequent cell death and/or significant loss of func - [4,5] Fresh aerial parts of J. Montana were collected from the Sinai acids, high oxygen consumption per unit of weight, high content since it presents: high content of peroxidizable unsaturated fatty Peninsula the plant was identified by Prof. Heba A. Elgizawy, Phar - macognosy department, Faculty of Pharmacy, October 6 University. Preparation of ethanolic extract in iron and ascorbate (LPO key ingredients) and a scarcity of an Air-dried aerial parts of the plant (1.5 kg) was crushed to coarse . In humans, the brain accounts for only a few percent of the tioxidant defense systems (e.g. GSH, GPx, CAT and vitamin E) [6- body weight, but it processes 20 % of basal oxygen consumption. A 11] The extract was concentrated under reduced pressure to yield vis- neuron uses much of oxygen via mitochondrial respiratory chain to powder and extracted exhaustively in a Soxhlet with 95% ethanol. cous mass. The ethanolic extract was kept in airtight containers in make ATP for maintaining low gradients . oC until the time of further use. [12,13] a deep freeze maintained at 4 agents in a multitude of disease states , most commonly Phytochemical screening Plant-derived polyphenols have been implicated as beneficial cancer, cardiovascular disease, and neurodegenerative disorders. A phytochemical analysis of aerial parts of J. Montana was con- [14,15] - lying these disorders and thus slowing the disease progression - Flavonoids are also able to counteract the neuronal injury under ducted for the detection of alkaloids, cardiac glycosides, flavonoids, terpenes . They have been shown to be effective at blocking oxidant- tannins, anthraquinones, saponins, volatile oil, coumarins and tri , although not via direct radical or [34]. [16,17] Animals oxidant-scavenging activity . induced neuronal injury [18] Adult albino rats weighing around 150 ± 10 gms, at the age of 10 [19,20] - weeks were purchased from National Cancer Institute, Cairo Uni- eas , including the Sinai Peninsula . The herb has a strong versity. They were acclimatized to animal house conditions. Ani- Jasonia Montana occurs in the Mediterranean and adjacent ar aromatic odor and is used in traditional medicine for diarrhea, [21] [22] stomachache, and chest diseases . were kept under constant environmental condition and observed mals were provided with standard diet and water ad libtum. Rats daily throughout the experimental work. Polyphenols [23] , and essential oils have been reported from Jasonia Mon- Experimental design [24], mono- and sesquiterpenes [25], flavonoids tana represent a promising potential species . These polyphe- [26] [27] nols are more potent antioxidants than vitamins C and E . Also, - [28] The rats were randomized and divided into five groups of six of the aerial parts of J. Montana was reported. In-vivo tests have mencement of experiment. The animals were divided into the fol- [29] rats each. Food was withdrawn 24 h and water 2 h before the com been conducted with Jasonia Montana to determine, for example, lowing groups (Table 1).

Citation: Mohammed A Hussein., et al. “Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia". Acta Scientific Nutritional Health

5.7 (2021): 27-37. Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

o C using a cooling ultra-centrifuge29 No. Groups Treatment Description gated at 10500 xg for 15 min at 4 I Normal control was used for the determination of Ca+2, Mg+2, Na+, K+-ATPase activi- (Sorvall comiplus T-880, Du Pont, USA), and the clear supernatant Received 5 mL saline orally, daily

for 14 days. II Disease control according to Tokeo and Sakanashi method - Received Scopolamine ties. The fourth aliquot used to extract and separate phospholipids hydrobromide (SCO) (3 mg/kg.b.w. [40]. Briefly, 1 mg of ho i.p), daily for 14 days [35]. mogenized tissue was treated with 950 μL of chloroform: methanol J. Montana ethanolic extract (150 III Treated group (A) Received SCO (3 mg/kg.b.w. i.p.) + (2:1, v: v) solution containing butylated hydroxytoluene (2 mM) and 50 μL of potassium phosphate buffer (100 mM, pH 7.4). The mg/kg b.w. orally) daily for 14 fifth aliquot was centrifuged at 5000 xg, and the supernatant was days [32]. analysis was run in parallel to test samples. The absorbance was donepezil used for the detection of β – Amyloid (Aβ1-42). Standard curve IV Treated group (B) Received SCO (3 mg/kg.b.w.i.p.) + measured in the multi-scan spectrum spectrophotometer (Thermo

hydrochloride (3 mg/kg, orally) were performed in triplicate. daily for 14 days [36]. Scientific, Multiskan GO) at optical density 450 nm. All the readings J. Montana ethanolic extract (150 V Treated group (C) Received SCO (3 mg/kg.b.w. i.p.) + Histological assessment

mg/kg b.w. orally) + donepezil 10% buffered formaldehyde solution for histological study. The hydrochloride (3 mg/kg, orally). The brain hippocampus was sliced, and pieces were fixed in Table 1: -

Groups of animals in the present study. fixed tissues were processed by automated tissue processing ma Plasma lipid profile estimation methods. Sections of 5 lm in thickness were prepared and then chine. Tissues were embedded in paraffin wax by conventional On 15th day, blood was collected from the retro-orbital vein of stained with hematoxylin and eosin for light microscopy analyses each animal and each sample was collected into heparinized tubes, according to the method of Bancroft and Steven . After that, the centrifuged, and plasma was used freshly for estimation of plasma sections were observed under the microscope for histopathologi- [41] cal changes, and their photomicrographs were captured. determined using commercially available kits (Asan and Young- plasma triglyceride, total cholesterol and HDL- cholesterol were dong Pharmaceutical Co., Korea) , respectively. Statistical analysis The results were expressed as means ± SD. Comparisons be- [37-39] Preparation of brain samples tween groups were performed using one-way analysis of variance (ANOVA). Differences between individual treatment groups were hippocampus were rapidly removed, extracted with saline at the Rats were dissected by cervical dislocation, and then brain - P < 0.05 and P < 0.01, and the statistical analyses were performed compared using Dunnett’s test. Statistical significance was set at mass-liquid ratio of 1:9, then the tissue homogenate was centri fuged at 3500 r/min for 10 min. The homogenate was divided into using SPSS software, version 15.0 (SPSS, Inc., Chicago, IL, USA). trichloroacetic acid and the obtained supernatant, after centrifuga- Results five aliquots. The first one was deproteinized with ice-cooled 12% - Table 2 showed that oral administration of scopolamine hydro- - - tion at 1000 xg, was used for the estimation of GSH level. The sec tant was used for estimation of acetylcholine esterase (AChE) and ond aliquot was centrifuged at 1000 xg and the resultant superna bromide (3 mg/kg.b.w. i.p) resulted in a significant increase in plas monoamine oxidase (MAO) activity as well as thiobarbaturic acid ma cholesterol and triglycerides as well as significantly decreased hippocampus phospholipids when compared to the normal control of plasma high-density lipoprotein-cholesterol (HDL-C) and brain - group (p < 0.01). Supplementation of J. Montana ethanolic extract reactive substances (TBARS), acetylcholine (ACh) and total protein - content s using immunoassay kits (Immuno-Biological Laborato ries ELISA kit). The third aliquot of brain homogenate was centrifu (150 mg/kg b.w.) and donepezil hydrochloride (3 mg/kg, orally)

Citation: Mohammed A Hussein., et al. “Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia". Acta Scientific Nutritional Health

5.7 (2021): 27-37. Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

the group that received scopolamine hydrobromide (p < 0.05). The30 effect of combination more pronounced than their administration individually or in combination resulted in a significant decrease in individuality. plasma cholesterol and triglycerides as well as significant increased plasma HDL-C and brain hippocampus phospholipids compared to Cholesterol Triglycerides HDL Phospholipids Groups Treatment Description (mg/dl) (mg/g tissue) (mg/dl) (mg/dl) Normal control I a a d e

78.55 ± 2.88 64.98 ± 4.23 34.87 ± 4.00 956.65 ± 12.66 II (5 mL saline) e d a a kg.b.w.) Scopolamine hydrobromide (SCO) (3 mg/ 132.98 ± 7.37 184.55 ± 9.48 21.54 ± 4.66 474.55 ± 18.70 III d c c c SCO (3 mg/kg.b.w. i.p.) + J. montana ethanolic - 96.06 ± 4.32 84.15 ± 5.85 30.76 ± 3.65 838.09 ± 16.48 IV extract (150 mg/kg b.w.) c c b b SCO (3 mg/kg.b.w.i.p.) + donepezil hydrochlo 110.56 ± 8.48 85.44 ± 6.73 25.40 ± 4.48 766.84 ± 11.47 ride (3 mg/kg.) V - b b d d SCO (3 mg/kg.b.w.) + J. montana ethanolic extract (150 mg/kg b.w.) + donepezil hydro 84.66 ± 4.98 73.11 ± 5.43 33.25 ± 2.98 879.55 ± 18.70 Table 2: chloride (3 mg/kg.). phospholipids in normal and experimental groups of mice. Levels of plasma cholesterol, triglycerides and cholesterol-high density lipoprotein (HDL) as well as brain hippocampus Values represent the mean ± SE (n = 6). Data shown are mean ± standard deviation of number of observations within each treatment. Data f

ollowed by the same letter are not significantly different at P ≤ 0.05. showed that oral administration of scopolamine hy- and AChE compared to the group that received scopolamine hydro-

Table 3 formula more pronounced than their administration individuality. drobromide (3 mg/kg.b.w. i.p) resulted in a significant increase in bromide (3 mg/kg.b.w. i.p) (p < 0.05). The effect of combination compared to the normal control group (p < 0.01). Supplementation brain hippocampus lipid TBARS and monoamine oxidase (MAO) - howed that oral administration of scopolamine hy- - Table 4 s of J. Montana ethanolic extract (150 mg/kg b.w.) and donepezil hy - drobromide (3 mg/kg.b.w. i.p) resulted in a significant increase drochloride (3 mg/kg, orally) individually or in combination result creased of brain hippocampus Ca+2, Mg+2 and Na+, K+-ATPase activ- monoamine oxidase (MAO) compared to the group that received in brain hippocampus β amyloid1-42 as well as significantly de ed in a significant decrease in brain hippocampus lipid TBARS and ity when compared to the normal control group (p < 0.01). Supple- - scopolamine hydrobromide (3 mg/kg.b.w. i.p) (p < 0.05). Also, oral - administration of scopolamine hydrobromide (3 mg/kg.b.w. i.p) re mentation of J. Montana ethanolic extract (150 mg/kg b.w.) and donepezil hydrochloride (3 mg/kg, orally) individually or in com sulted in a significant decrease in brain hippocampus cetylcholine - esterase (AChE) activity, compared to the normal control group (p bination resulted in a significant decrease in brain hippocampus β (ACh), reduced glutathione (GSH) contents as well as acetylcholine pus Ca+2, Mg+2 and Na+, K+-ATPase activity when compared to the amyloid1-42 as well as significantly increased of brain hippocam - < 0.01). Supplementation of J. Montana ethanolic extract (150 mg/ (p < 0.05). The effect of combination more pronounced than their group that received scopolamine hydrobromide (3 mg/kg.b.w. i.p) kg b.w.) and donepezil hydrochloride (3 mg/kg, orally) individu administration individuality. ally or in combination resulted in a significant increase in GSH, Ach,

Citation: Mohammed A Hussein., et al. “Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia". Acta Scientific Nutritional Health

5.7 (2021): 27-37. Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

TBARS GSH ACh Ach E MAO 31 Groups Treatment Description (nmole MDA/ (µM/mg (mM/mg (U/mg (U/mg pro- mg protein) protein) protein) protein) tein) Normal control I a d c a a

Scopolamine hydrobromide (SCO) 15.48 ± 1.22 0.52 ± 0.031 8.54 ± 0.54 0.45 ± 0.05 23.87 ± 2.15 II (5 mL saline) d 0.15 ± 0.025 a a e d

32.45 ± 3.66 4.33 ± 0.65 1.43 ± 0.13 43.76 ± 4.56 III (3 mg/kg.b.w.) b c b d ab SCO (3 mg/kg.b.w. i.p.) + J. montana 18.66 ± 2.04 0.43 ± 0.05 7.54 ± 0.47 0.85 ± 0.08 25.90 ± 3.76 IV ethanolic extract (150 mg/kg b.w.) 22.15 ± 2.71c b b c c SCO (3 mg/kg.b.w.i.p.) + donepezil 0.39 ± 0.04 7.10 ± 0.54 0.94 ± 0.05 29.48 ± 2.87 hydrochloride (3 mg/kg) V ab d c b b SCO (3 mg/kg.b.w.) + J. montana ethanolic extract (150 mg/kg b.w.) + 17.46 ± 1.95 0.51 ± 0.04 8.09 ± 0.36 0.71 ± 0.09 26.66 ± 2.37 Table 3: donepezil hydrochloride (3 mg/kg). contents as well as acetylcholine esterase (AChE), monoamine oxidase (MAO) activity in normal and experimental groups of mice. Levels of brain hippocampus thiobarbaturic acid reactive substances (TBARS), acetylcholine (ACh), reduced glutathione (GSH) Values represent the mean ± SE (n = 6). Data shown are mean ± standard deviation of number of observations within each treatment. Data followed by the same letter are not significantly different at P ≤ 0.05.

+2 Ca -ATPase Mg+2-ATPase (U/ Na+, K+-ATPase (U/ Aβ1-42 Groups Treatment Description (U/mg protein) mg protein) mg protein) (pg/mg protein) Normal control I e 117.50 ± 10.77d d a

Scopolamine hydrobromide (SCO) 164.87 ± 6.87 132.98 ± 18.75 47.87 ± 5.26 II (5 mL saline) a a a d

- 76.45 ± 5.33 93.89 ± 5.32 64.09 ± 8.44 135.04 ± 10.45 (3 mg/kg.b.w.) III c c c b kgSCO b.w.) (3 mg/kg.b.w. i.p.) + J. mon tana ethanolic extract (150 mg/ 143.00 ± 12.86 112.52 ± 8.00 124.37 ± 10.66 55.26 ± 4.37 IV b b b c SCO (3 mg/kg.b.w.i.p.) + donepezil 122.65 ± 12.90 106.88 ± 11.25 110.87 ± 9.80 63.53 ± 3.77 hydrochloride (3 mg/kg. V d cd c a b.w.)SCO (3 + donepezilmg/kg.b.w.) hydrochloride + J. montana ethanolic extract (150 mg/kg 150.66 ± 11.54 115.40 ± 13.29 128.73 ± 14.63 46.75 ± 4.88

Table 4: (3 mg/kg). +2, Mg+2 and Na+, K+-ATPase activity as well as β amyloid1-42 level in normal and experimental groups of rats. Levels of brain hippocampus Ca Values represent the mean ± SE (n = 6). Data shown are mean ± standard deviation of number of observations within each treatment. Data followed by the same letter are not significantly different at P ≤ 0.05.

Citation: Mohammed A Hussein., et al. “Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia". Acta Scientific Nutritional Health

5.7 (2021): 27-37. Jasonia Montana; A Promising Therapeutic Agent to Attenuate Neurological Disorders Associated with SCO-induced Dementia

Histopathology examination Here we investigated thiobarbaturic acid reactive substances32 Histopathological examination of brain hippocampus sections - tents as well as acetylcholine esterase (AChE), monoamine oxidase of the normal group (I). On the other hand, in the brain hippocam- (TBARS), acetylcholine (ACh), reduced glutathione (GSH) con pus of SCO-treated control group (II), histological examination (MAO) and comparatively studied the effects of different fatty acid composition of PC on brain function. Scopolamine, a traditional anti-cholinergic drug, is reported to have the capability of impair- showed amyloid plaques seen in Alzheimer (arrow). - - short-term memory . Also, J. Montana ethanolic extract (150 mg/kg b.w.) and donepe ing spatial learning and memory and influencing the formation of zil hydrochloride (3 mg/kg, orally) treated group (III and IV) dem [48] also noted in brain hippocam