|Year : 2020 | Volume
| Issue : 3 | Page : 120-127
Therapeutic Evaluation of Thymoquinone in the Intracerebroventricular Injection of L-Cysteine Induced Vascular Dementia in Rats
Narahari Rishitha1, Arunachalam Muthuraman2, Chidambaram Saravanababu1
1 Department of Pharmacology, JSS College of Pharmacy, JSS Academy of Higher Education and Research, Mysuru, Karnataka, India
2 Department of Pharmacology, JSS College of Pharmacy, JSS Academy of Higher Education and Research, Mysuru, Karnataka; Pharmacology Unit, Faculty of Pharmacy, AIMST University, Semeling, Bedong, Kedah Darul Aman, Malaysia, India
|Date of Submission||07-Jan-2020|
|Date of Decision||02-Feb-2020|
|Date of Acceptance||03-Mar-2020|
|Date of Web Publication||20-Aug-2020|
MPharm, PhD Arunachalam Muthuraman
Pharmacology Unit, Faculty of Pharmacy, AIMST University, Semeling, 08100 Bedong, Kedah, Darul Aman, Malaysia
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Introduction: Worldwide, dementia is one of the leading causes of death. The major type of dementia disorders is Alzheimer disease (AD) and vascular dementia (VaD). The progress of VaD is due to the lack of blood flow; and the accumulation of metabolic neurotoxin in the nervous system. In the present study, VaD was induced by intracerebroventricular injection of L-Cysteine (L-Cys) in rats. Methods: Thymoquinone (TQ; 5 and 10 mg/kg; p.o.) was administered for 5 consecutive days. Donepezil (10 mg/kg; p.o. 5 consecutive days) was used as a reference control. Neurocognitive functions were tested at different time intervals. Furthermore, L-Cys induced biochemical changes were also estimated in plasma and hippocampus tissue samples. Results: Treatment of TQ ameliorates the L-Cys induced cognitive impairments along with biochemical changes as similar to donepezil treatment. Conclusion: Hence, the TQ may be a newer medicine for dementia disorder due to its anti-oxidant, anti-lipid peroxidative, anti-inflammatory actions; regulation of cholinergic neurotransmission; and reduction of metabolic accumulation of neurotoxin.
Keywords: Acetylcholinesterase, Alzheimer’s disease, cognitive function, donepezil, homocysteine, Vascular dysfunction
|How to cite this article:|
Rishitha N, Muthuraman A, Saravanababu C. Therapeutic Evaluation of Thymoquinone in the Intracerebroventricular Injection of L-Cysteine Induced Vascular Dementia in Rats. Int J Nutr Pharmacol Neurol Dis 2020;10:120-7
|How to cite this URL:|
Rishitha N, Muthuraman A, Saravanababu C. Therapeutic Evaluation of Thymoquinone in the Intracerebroventricular Injection of L-Cysteine Induced Vascular Dementia in Rats. Int J Nutr Pharmacol Neurol Dis [serial online] 2020 [cited 2020 Oct 26];10:120-7. Available from: https://www.ijnpnd.com/text.asp?2020/10/3/120/292678
| Introduction|| |
Dementia is a chronic progressive neurocognitive disorder. Numerous ethnological factors are causing dementia-like aging, smoking, alcohol, infection, and disease. Sometimes, it occurs with endogenous factors like the formation of uremia, accumulation of cholesterol and glucose, synthesis of metabolic waste products like urea, creatinine, guanidine, methyl guanidine, homocysteine, and cysteinyl catechols in the nervous system. The major type of dementia disorders is Alzheimer disease (AD) and vascular dementia (VaD). The progress of VaD is due to the lack of blood flow; and the accumulation of metabolic neurotoxin in the nervous system. The alteration of blood flow occurs with narrow blood vessels, deposition of cholesterol in a cerebral artery, and the formation of thrombosis. These events are occurring with changes of ion channels and neurotransmitter regulation in cerebral blood vessels leads to the abnormal blood-brain barrier (BBB) and nerve-blood barrier (NBB) functions. This is an initial pathological event followed by it accelerates the calcium dys-homeostasis, alteration of the endogenous anti-oxidant system and changes of lipid-derived radicals by the peroxidation process leads to alter the cellular, mitochondrial and nuclear membrane dysfunctions. Furthermore, it also alters the normal mitochondrial and nuclear DNA; and neuronal cytoskeletal proteins. Homocysteine is identified as a major culprit in DNA damage in the neurovascular system leads to cause cognitive dysfunctions. The reductions of homocysteine levels occur by blocking the action of adenosine conversion or enhance the conversion of L-methionine or L-cysteine (L-Cys).
Experimentally, the chronic administration of L-methionine (1.7 g/kg, p.o. for 32 days) enhances vascular dementia by the accumulation of homocysteine., In addition, it also enhances the abnormalities of cerebral vascular reactivity via nitrite and superoxide radicals; and endothelial dysfunction. Various amino acids are contributed to the regulation of neurological functions. And it’s also involved in neurological disorders. The concentration and duration of amino acids are responsible for the neuroprotective and neurotoxic effects. Some of the amino acids such as glutamate, aspartate, cysteine, and methionine are enhanced neurodegeneration via activating NMDA receptor, kynurenine and homocysteine pathways.,, Some studies reported that neurotoxic effects of L-Cys are due to alteration of ADP-ribosylation, activation of excitatory amino acid receptors that is, N-methyl-D-aspartate (NMDA), potentiating of Ca2+ influx, and formation of endogenous cysteinyl catechols. However, the role of L-Cys in the pathogenesis of vascular dementia is not explored yet.
Current conventional medicines like donepezil, galantamine, and rivastigmine are partially used for the treatment of cognitive disorders including AD and VaD., However, it relieves the symptoms partially and it produces the various unwanted side effects like liver damage; hematological and psychological changes; and reproductive toxicity.,, However, the specific medicines with more potent and high efficacy are not explored yet. Hence, the newer medicines are required to manage the neurovascular disorders including VaD. Thymoquinone (TQ) is a primary constituent in the oil of Nigella sativa seeds and it has multiple biological actions on various systems of the body., Experimentally, it is proven that it has antioxidant, anti-inflammatory, chemo-preventive, antitumor, anticancer, immunomodulatory, antimicrobial, anti-parasitic, hypoglycemic, antihypertensive, and anti-asthmatic effects.,,,, In addition, it also produces neuroprotective action against spinal cord ischemia and cerebral ischemia-reperfusion injury., Further, it also shown prevents the glutamate and Aβ1–42-induced neurotoxicity., Some reports also attenuate the memory-enhancing effects against lipopolysaccharide-induced Alzheimer’s disease. Therefore, the present study designed to investigate the role of TQ in intracerebroventricular (i.c.v.) administration of L-Cys induced VaD in rats.
| Materials and Methods|| |
Wistar rat (150 to 180 g) was used in the present research work. Animals fed with standard laboratory diet (procured from Adita Biosys private limited, Tumkur, Karnataka, India) and allow to access free water ad libitum. The 12 hours light/dark cycles were maintained in the animal house and the experimental laboratory. All behavioral observation was made in a semi-soundproof laboratory. The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC No.: 301/2018). The care of the animals was followed as per the guidelines of ‘Committee for the Purpose of Control and Supervision of Experiments on Animals’ (CPCSEA), Ministry of Environment and Forest, Government of India (Reg. No. 155/PO/Re/S/1999/CPCSEA; dated 11/09/2015).
Drugs and chemicals
Thymoquinone is obtained from TCI, India. Acetylthiocholine and 1, 1, 3, 3-tetra methoxy propane were procured from S.D. Fine Chemicals, Mumbai. L-Cys, Folin-Ciocalteu’s Phenol reagent, 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), reduced glutathione (GSH) and bovine serum albumin (BSA) were purchased from Sisco Research Laboratories Pvt. Ltd., Mumbai. Homocysteine obtained from Sigma Aldrich, Mumbai. All other reagents were used as an analytical grade. All the solutions were freshly prepared before use.
Induction of vascular dementia by i.c.v. administration of L-Cys
The i.c.v. administration of L-Cys was made as a described method of Yamamoto and Mohanan. Briefly, rats were anesthetized with anesthetic agent (50 mg/kg of ketamine, i.p.,; & 10 mg/kg of xylazine, i.p.) and i.c.v. injection of L-Cys was made with a hypodermic needle (0.4 mm external diameter of Hamilton microlitre syringe). L-Cys was dissolved in freshly prepared artificial cerebrospinal fluid (ACSF; 25 mg/ml) solution. The 10 µl of L-Cys was administered bilaterally to i.c.v. location of the rat brain with the help of stereotactic apparatus. The location of i.c.v. injection in the rat brain was coordinated with the help of stereotactic atlas of the rat brain. The coordinates of rat i.c.v. i.e., 0.8 mm posterior to the bregma, 1.8 mm lateral to the sagittal suture and 3.6 mm beneath the cortical surface were used.
All rats were acclimatized in laboratory conditions for a period of one week prior to the initiation of behavioral assessment and experimental protocol. Animal distribution in different groups was randomized based on their stratified body weight. Five groups were employed in present research work. Each group was comprised of six animals (n = 6). Group I: Normal control group; Group II: L-Cysteine (L-Cys) group; Group III and IV: L-Cys + thymoquinone (TQ; 5 and 10 mg/kg; p.o.) for five consecutive days; Group V: L-Cys + donepezil (10 mg/kg; p.o.) for 5 consecutive days treated group. On the last day of the study protocol, all the animals were used for the collection of blood samples by the tail vein method under mild ether anesthetic conditions. The plasma was separated and stored in defreezer (−20ᵒC). Thereafter, animals were sacrificed by cervical dislocation and the brain was isolated immediately. Then the hippocampus part of the brain was separated for the estimation of tissue biomarker changes.
All the rats were engaged in different memory function tests. The learning and memory function tests were evaluated by using the Morris water maze test; elevated pulse maze test; lateral push test (LPT) and social interaction tests. The movements of the animal during the cognitive assessment were recorded with a USB camera (12 Megapixel, Intex products, India).
Assessment of neurocognitive function by Morris water maze (MWM) test
MWM test is one of the established methods for the assessment of spatial learning and memory behavior of rodents. MWM test was described by Morris. MWM was consists of a 150 cm diameter of the water pool with the division of four equal quadrants (Q1 to Q4). The platform (10 × 10 × 30 cm) platform was placed in the target quadrant (Q4). Water was filled to the 32 cm height. The rat was trained in the MWM test apparatus after five days of i.c.v. L-Cys administration. Learning behavior of the rat was assessed four consecutive days i.e., day 6 to day 9. During this period, escape latency time (ELT, the time taken to reach the platform) was noted. On the tenth day, the memory test was performed without the platform with opaque (milk was added) water. Time spent in the target quadrant (TSTQ) was noted as a memory function.
Assessment of neurocognitive function by elevated pulse maze (EPM) test
EPM test was used for the assessment of memory as well as anxiety behavior in rodents. Short-term spatial memory was performed by the EPM test as a described method of Kawaguchi et al. The animals were placed in the corner of the open arm and allow to move the end of the closed arm. Transfer latency (TL, the time is taken to reach the end of the closed arm) was noted as memory function. TL reading was noted on the 6th day of the experimental protocol as a learning (training) period. TL reading on 7th day in the EPM test was noted as memory assessment.,
Assessment of grip strength by lateral push test (LPT)
LPT was used for the assessment of grip strength as an indication of motor-coordinated function. LPT test was described by Petullo et al. with a slight modification of Schaar et al. The lateral push stimuli were applied five times on each side of the animal body. The test was performed at the end of the experimental protocol. The percentage resistance against the lateral push (pressure) was calculated for the assessment of grip strength based on a number of stimuli and the number of resistant response.
Assessment of social interaction test (SIT)
SIT was used for the assessment of the psychosocial behavior of rodents and this method was described by Varlinskaya and Spear with a slight modification of Kaidanovich-Beilin et al. All the experimental animals were placed individually in the right diagonal corner, and the face was kept towards the wall surface. The assessment of social interaction was noted by the time taken for complete interaction with unfamiliar (circular box) animals. Between each animal testing, circular chamber and the square chamber were cleaned with non-odorant Bio Green Solutions (a mixture of Natural starch; vegetable oil; and vegetable waste) to avoid the biases of the olfactory cue. Each chamber boxes was allowed exchanging air to avoid the stress on the animals. All SIT were performed between 9:00 to11:00 AM with room lighting of 650 Lux.
Estimations of plasma biomarker changes
The blood samples were collected at the end of the study protocol via the tail vein method as described by Vogel and Vogel. Briefly, rat-tail vein was punctured and blood samples were collected in centrifugal tubes. Centrifugal tube was prefilled with 100 µl of anti-coagulant i.e., 11 % w/v of sodium citrate. The centrifugal tubes were centrifuged at 3000 rpm for 15 minuteswith15 ᵒC.
Estimation of plasma homocysteine (HCy) level
The plasma samples were used for the assessment of homocysteine (HCy) estimation using the ultraviolet spectrophotometer method as described by Wang et al. The changes of absorbance in test tube solution were noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 510 nm wavelength. The standard plot was prepared with 1 to 10 µmol of HCy concentration to calculate the concentration of plasma HCy levels. The standard HCy was prepared with 70 percentage of hydro-alcoholic preparation. The results were expressed as μmol per liter.
Estimation of plasma nitrite level
The level of plasma nitrite changes was estimated by using Griess reagent as a described method of Green et al. The developed pink color chromogen in supernatant solution and their absorbance change was noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 540 nm wavelength. The standard curve of sodium nitrite was plotted with 5 to 50 μmol concentration to calculate the concentration of plasma nitrite levels. The results were expressed as μmol per liter.
Estimations of tissue biomarker changes
After completion of blood sample collection, all rats were sacrificed by cervical dislocation. The brain was isolated immediately; then, the brain was dissected for the separation of the hippocampus. All the tissue was homogenated with a phosphate buffer with pH − 7.4. In addition, the supernatant was separated by centrifugation process at 3000 rpm for 15 min with 15ᵒC. Then, the hippocampus supernatant was used for the various biochemical estimations such as acetylcholinesterase (AChE) activity; reduced glutathione (GSH); thiobarbituric acid reactive substances (TBARS); myeloperoxidase (MPO) activity levels. In addition, HCy levels also estimated in hippocampus samples.
Estimation of AChE level
AChE activity level was estimated by a spectroscopic method as described by Ellman et al. The change of absorbance was noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 420 nm wavelength. These readings were used for the further calculation of AChE activity levels in a tissue sample. The results were expressed as µM of acetylthiocholine hydrolysis per milligram of protein per minute.
Estimation of GSH level
GSH level was estimated as a described method of Ellman. The developed yellow color chromogen changes of absorbance were noted by spectrophotometer (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 412 nm wavelength. The results were expressed as µmol of GSH / mg of protein.
Estimation of TBARS level
TBARS level of brain samples was estimated as a described method of Ohkawa et al. The clear pink color chromogen of the supernatant was collected. The change of absorbance was noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 535 nm wavelength. The results were expressed as nmol per mg of protein.
Estimation of myeloperoxidase (MPO) activity level
MPO levels were estimated as a described method of Grisham et al. and Patriarca et al. The change of absorbance was noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 460 nm wavelength. The results were expressed as myeloperoxidase activity units per milligram of protein at a one-minute duration.
Estimation of HCy level
HCy levels were estimated as a described method of Wang et al. after the reaction developed solution color was changed from bright yellow to the brownish-orange color chromogen. The changes of absorbance were noted by spectrophotometrically (DU 640B Spectrophotometer, Beckman Coulter Inc., CA, USA) at 510 nm wavelength. The standard plot was prepared with 1 to 10 µmol solution of HCy. The standard HCy was prepared with 70 percentage of hydro-alcoholic preparation. The results were expressed as µmol per mg of protein.
Estimation of total protein level
Total protein level levels were estimated as a described method of Lowry et al. The change of absorbance was noted by spectrophotometrically (DU 640B, UV-Spectrophotometer, Beckman Coulter Inc., CA, USA) at 750 nm wavelength. These readings were used for the further integrated calculation of total protein levels versus AChE, GSH, TBARS, MPO and HCy levels in tissue samples. The results were expressed as mg of protein per ml of supernatant.
All the results were expressed as mean ± standard deviation (SD). Data obtained from behavioral tests were statistically analyzed using two-way analysis of variance (ANOVA) and data of biomarkers changes i.e., AChE activity, GSH, TBARS, MPO, and HCy levels were analyzed using one way ANOVA followed by Tukey’s multiple range tests were applied for Post-hoc analysis by using Graph pad prism version-5.0 software. A probability value of P < 0.05 was considered statistically significant.
| Results|| |
Effect of TQ on learning and memory behavior in MWM test
The normal animal in acquisition trial days that is, day 9 ELT value significantly (P < 0.05) decreases when compared to day 6 ELT value. It indicates that normal rats possess significant learning abilities. In the retrieval test, the normal animal was shown a significant (P < 0.05) increase in the TSTQ (i.e., Q4 quadrant) value when compared to the Q1 quadrant value. The administration of L-Cys of rat brain produced a significant decrease in TSTQ (i.e., Q4) when compared to the sham group. It indicates an index of memory retrieval function. The treatment of TQ (5 and 10 mg/kg; p.o. for 5 consecutive days) is shown a significant abolishing effect in L-Cys induced reduction of TSTQ value dose-dependent manner. Hence, TQ produced the improvement of memory function in rats against the L-Cys induced brain damage. Similarly, the treatment of reference compound that is, donepezil (10 mg/kg; p.o. for 5 consecutive days) also produced the significant raising of TSTQ value [Figure 1]a and 1b.
|Figure 1 Effect of TQ on learning and memory behavior in the MWM test. Digits in parenthesis indicate dose mg/kg. Data were expressed as mean ± SD, n = 6 rats per group. αP < 0.05 when compared to Q1 normal group. βP < 0.05 when compared to Q4 normal group. δP < 0.05 when compared to L-Cys group. Abbreviation: ELT, escape latency time; TSTQ, time spent in target quadrant; L-Cys, L-Cysteine; and TQ, thymoquinone.|
Click here to view
Effect of TQ on learning and memory behavior in the EPM test
The administration of L-Cys produced significant (P < 0.05) increases the transfer latency (TL) time when compared to the normal group. The treatment of TQ (10 mg/kg; p.o.) significantly attenuates the L-Cys induced raising of TL duration. It indicates that L-Cys has memory enhancement effects in rats against L-Cys induced neuronal damage. Whereas, the treatment of TQ (5 mg/kg) is not shown any changes in the EPM test. The similar effect also observed in reference compound that is, donepezil (10 mg/kg; p.o.) treatment group [Figure 2].
|Figure 2 Effect of TQ on learning and memory behavior in the EPM test. Digits in parenthesis indicate dose mg/kg. Data were expressed as mean ± SD, n = 6 rats per group. αP < 0.05 when compared to the normal group, and βP < 0.05 when compared to the L-Cys group. Abbreviation: L-Cys, L-Cysteine; and TQ, thymoquinone.|
Click here to view
Effect TQ on grip strength pattern in LPT
The administration of L-Cys produced significant (P < 0.05) reduction of percentage grip strength against lateral push stimuli when compared to the sham control group. The treatment of TQ (10 mg/kg; p.o.) significantly attenuates L-Cys induced changes of grip strength. It indicates that TQ (80 mg/kg) treatment possess the improvement of grip strength ability with neuromuscular coordinated function in rat against L-Cys induced neuronal damage. Whereas, the treatment of TQ (5 mg/kg) is not shown any changes in LPT against L-Cys in the rat. The similar effect also observed in reference compound that is, donepezil (10 mg/kg; p.o.) treatment group [Figure 3].
|Figure 3 Effect TQ on grip strength pattern in LPT. Digits in parenthesis indicate dose mg/kg. Data were expressed as mean ± SD, n = 6 rats per group. αP < 0.05 when compared to the normal group, and βP < 0.05 when compared to the L-Cys group. Abbreviation: L-Cys, L-Cysteine; and TQ, thymoquinone.|
Click here to view
Effect of TQ on psychosocial behavior by SIT
The administration of L-Cys was produced significant (P < 0.05) decrease the duration of interaction when compared to the sham control group. Treatment of TQ (10 mg/kg; p.o.) significantly attenuates the L-Cys induced changes in social interaction time. It indicates that TQ (10 mg/kg) treatment possess the improvement of psychosocial behavior ability within rat against L-Cys induced neuronal damage. Whereas, treatment of TQ (5 mg/kg) is not shown any changes in SIT against L-Cys in the rat. The similar effect also observed in reference compound that is, donepezil (10 mg/kg; p.o.) treatment group [Figure 4].
|Figure 4 Effect of TQ on psychosocial behavior by SIT. Digits in parenthesis indicate dose mg/kg. Data were expressed as mean ± SD, n = 6 rats per group. αP < 0.05 when compared to the normal group, and βP < 0.05 when compared to the L-Cys group. Abbreviation: L-Cys, L-Cysteine; and TQ, thymoquinone.|
Click here to view
Effect of TQ on plasma biochemical changes
The administration of L-Cys was produced significant (P <0.05) rising of HCy and nitrite levels when compared to the sham group. The treatment of TQ (5 and 10 mg/kg; p.o.) significantly attenuated the L-Cys induced above biochemical changes in a dose-dependent manner. It indicates that TQ regulates the accumulation of metabolic toxin and endothelial-derived vaso-relaxing factors (EDRF: Nitric oxide) leads to produce the neurovascular protection in rat brain. The similar changes also observed in reference compound that is, donepezil (10 mg/kg; p.o.) treatment group [Table 1].
Effect of TQ on tissue biochemical changes
The administration of L-Cys was produced significant (P < 0.05) rising of AChE activity; TBARS; MPO and HCy along with a decrease in GSH levels when compared to the sham group. The treatment of TQ (5 and 10 mg/kg; p.o.) significantly attenuated the L-Cys induced above biochemical changes in a dose-dependent manner. It indicates that TQ produces the anti-oxidative; anti-lipid peroxidative; anti-inflammatory and regulation of cholinergic neurotransmission as well as metabolic toxin actions. The similar changes also observed in reference compound that is, donepezil (10 mg/kg; p.o.) treatment group [Table 2].
| Discussion|| |
The current results showed L-Cys causes the potential neurovascular damage leads to impairment of learning and memory patterns in the rat. In addition, L-Cys alters the normal biochemical levels in plasma and hippocampus tissue samples that is, rising the AChE activity, TBARS, MPO, and HCy levels; and reduce the GSH level. The administration of TQ (5 and 10mg/kg; p.o.) significantly (P < 0.05) ameliorates the L-Cys induced cerebrovascular injury, cognitive impairments, and neuroinflammation in rats. It is also comparable with the reference compound that is, donepezil (10 mg/kg; p.o.) treatment.
L-Cys is causing the potential neurotoxic action via multiple pathophysiological pathways similar to that of L-methionine. L-methionine and L-Cys have a common pathophysiological mechanism that is, the elevation of HCy. In addition, it has additional interactive action with the excitatory amino acid receptor, especially the NMDA receptor with the opening of cellular Ca2+ion channels. Furthermore, HCy also enhances the synthesis and release of glutamate and aspartate amino acid, which leads to enhance neuronal membrane potential. Concurrently, the NMDA and HCy associated activation of neuronal membrane potential produces the neuronal excitation and loss of neuronal plasticity.
Hence, it causes memory impairments. Paradoxically, free radicals and lipid peroxidation also involve the activation of calcium and NMDA receptor-associated ion channels lead to cause memory dysfunction. Numerous study reports that free radical scavenger, anti-lipid peroxidation, and anti-inflammatory molecules were documented to produce the neuroprotection and memory-boosting effects. In addition, various neurotoxic chemicals like L-Cys induce neurodegeneration via oxidative, lipid peroxidative and inflammatory pathways. In addition, biomarkers of these pathways that is, TBARS, GSH, and MPO also reported in neurotoxic associated dementia disorders. The modulators of these pathways attenuate neurodegeneration and cognitive impairments. Moreover, the hallmark of cognitive disorders changes in acetylcholine levels in the brain tissue. It regulates the neuronal functions. Whereas, the degradation of acetylcholine by AChE were accelerated the cognitive dysfunction including vascular dementia.In the present research work, also shown similar biochemical changes in L-Cys treated animals. TQ treated animals were reduced the L-Cys induced activation of AChE activity levels. The literature report revealed that the treatment of TQ shown potent antioxidant, anti-inflammatory, and anti-lipid peroxidative agents., In addition, it is also documented to produce immunomodulatory and neurotransmitter regulatory actions. Further, it showed inhibitory action of α7 nicotinic acetylcholine receptors (α7 nAchRs) and acetylcholinesterase activity. Experimental research reports also evidenced that, TQ prevents the glutamate and Aβ1–42-induced neurotoxicity., TQ also reported that it attenuates the lipopolysaccharide-induced memory impairment and symptoms of Alzheimer’s disease. The present research work also showed similar results in behavioral and biochemical evaluation. Hence, it concluded that TQ may be a future medicine for metabolic and neurotoxic associated neurovascular disorders due to its potential anti-oxidative, anti-lipid peroxidative, anti-inflammatory, and acetylcholinesterase inhibitory actions. However, more extensive research studies are required to explore the complete therapeutic potency, efficacy, and safety of TQ in various pathophysiological conditions of neurovascular disorders including vascular dementia.
The authors are thankful to the College of Pharmacy, JSS Academy of Higher Education and Research, Mysuru-570 015, Karnataka, India, and Pharmacology Unit, Faculty of Pharmacy, AIMST University, Semeling, 08100-Bedong, Kedah Darul Aman, Malaysia for supporting this study and providing technical facilities for this work.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Duong S, Patel T, Chang F. Dementia: What pharmacists need to know. Can Pharm J 2017;150:118-29.
Bednarska-Makaruk M, Graban A, Sobczyńska-Malefora A, Harrington DJ, Mitchell M, Voong K, Dai L, Łojkowska W, Bochyńska A, Ryglewicz D, Wiśniewska A, Wehr H. Homocysteine metabolism and the associations of global DNA methylation with selected gene polymorphisms and nutritional factors in patients with dementia. Exp Gerontol 2016;81:83-91.
Raz L, Knoefel J, Bhaska K. The neuropathology and cerebrovascular mechanisms of dementia. J Cereb Blood Flow Metab 2016;36:172-86.
Prakash R, Carmichael ST. Blood-brain barrier breakdown and neovascularization processes after stroke and traumatic brain injury. Curr Opin Neurol 2015;28:556.
Xiao M, Zhong H, Xia L, Tao Y, Yin H. Pathophysiology of mitochondrial lipid oxidation: role of 4-hydroxynonenal (4-HNE) and other bioactive lipids in mitochondria. Free Radic Biol Med 2017;111:316-27.
El-Dessouki AM, Galal MA, Awad AS, Zaki HF. Neuroprotective effects of simvastatin and cilostazol in L-Methionine-Induced vascular dementia in rats. Mol Neurobiol 2017;54:5074-84.
Mangat GS, Jaggi AS, Singh N. Ameliorative effect of a selective endothelin ETA receptor antagonist in rat model of L-methionine-induced vascular dementia. Korean J Physiol Pharmacol 2014;18:201-9.
Martin-Hernandez D, Tendilla-Beltran H, Madrigal JLM, Garcia-Bueno B, Leza JC, Caso JR. Chronic mild stress alters kynurenine pathways changing the glutamate neurotransmission in frontal cortex of rats. Mol Neurobiol 2019;56:490-501.
Kohl JB, Mellis AT, Schwarz G. Homeostatic impact of sulfite and hydrogen sulfide on cysteine catabolism. Br J Pharmacol 2019;176:554-70.
Bajrektarevic D, Nistri A. Ceftriaxone-mediated upregulation of the glutamate transporter GLT-1 contrasts neurotoxicity evoked by kainate in rat organotypic spinal cord cultures. Neurotoxicology 2017;60:34-41.
Montine T, Picklo M, Amarnath V, Whetsell Jr. W, Graham DG. Neurotoxicity of endogenous cysteinyl catechols. Exp Neurol 1997;148:26-33.
Farooq MU, Min J, Goshgarian C, Gorelick PB. Pharmacotherapy for vascular cognitive impairment. CNS Drugs 2017;31:759-76.
Kandiah N, Pai MC, Senanarong V, Looi I, Ampil E, Park KW, Karanam AK, Christopher S. Rivastigmine: the advantages of dual inhibition of acetylcholinesterase and butyrylcholinesterase and its role in subcortical vascular dementia and Parkinson’s disease dementia. Clin Interv Aging 2017;12:697-707.
Li DD, Zhang YH, Zhang W, Zhao P. Meta-analysis of randomized controlled trials on the efficacy and safety of donepezil, galantamine, rivastigmine, and memantine for the treatment of Alzheimer’s disease. Front Neurosci 2019;13:472.
Tan CC, Yu JT, Wang HF, Tan MS, Meng XF, Wang C, Jiang T, Zhu XC, Tan L. Efficacy and safety of donepezil, galantamine, rivastigmine, and memantine for the treatment of Alzheimer’s disease: a systematic review and meta-analysis. J Alzheimers Dis 2014;41:615-31.
Zhang N, Gordon ML. Clinical efficacy and safety of donepezil in the treatment of Alzheimer’s disease in Chinese patients. Clin Interv Aging 2018;13:1963-970.
Radad K, Hassanein K, Al-Shraim M, Moldzio R, Rausch WD. Thymoquinone ameliorates lead-induced brain damage in Sprague Dawley rats. Exp Toxicol Pathol 2014;66:13-7.
Fanoudi S, Alavi MS, Hosseini M, Sadeghnia HR. Nigella sativa and thymoquinone attenuate oxidative stress and cognitive impairment following cerebral hypoperfusion in rats. Metab Brain Dis 2019;34:1001-10.
Shaterzadeh-Yazdi H, Noorbakhsh MF, Samarghandian S, Farkhondeh T. An overview on renoprotective effects of thymoquinone. Kidney Dis 2018;4:74-82.
Noorbakhsh MF, Shaterzadeh-Yazdi H, Hayati F, Samarghandian S, Farkhondeh T. Protective effects of thymoquinone on pulmonary disorders in experimental studies. Tanaffos 2018;17:211-22.
Jakaria M, Cho DY, EzazulHaque M, Karthivashan G, Kim IS, Ganesan P, Choi DK. Neuropharmacological potential and delivery prospects of thymoquinone for neurological disorders. Oxid Med Cell Longev 2018;2018:1209801.
Oskouei Z, Akaberi M, Hosseinzadeh H. A glance at black cumin (Nigella sativa
) and its active constituent, thymoquinone, in ischemia: a review. Iran J Basic Med Sci 2018;21:1200-9.
Mahmoud YK, Abdelrazek HMA. Cancer: Thymoquinone antioxidant/pro-oxidant effect as potential anticancer remedy. Biomed Pharmacother 2019;115:108783.
Fouad IA, Sharaf NM, Abdelghany RM, El Sayed N. Neuromodulatory effect of thymoquinone in attenuating glutamate-mediated neurotoxicity targeting the amyloidogenic and apoptotic pathways. Front Neurol 2018;9:236.
Alhibshi AH, Odawara A, Suzuki I. Neuroprotective efficacy of thymoquinone against amyloid beta-induced neurotoxicity in human induced pluripotent stem cell-derived cholinergic neurons. Biochem Biophys Rep 2019;17:122-6.
Bargi R, Asgharzadeh F, Beheshti F, Hosseini M, Sadeghnia HR, Khazaei M. The effects of thymoquinone on hippocampal cytokine level, brain oxidative stress status and memory deficits induced by lipopolysaccharide in rats. Cytokine 2017;96:173-84.
Yamamoto HA, Mohanan PV. In vivo and in vitro effects of melatonin or ganglioside GT1B on L-cysteine-induced brain mitochondrial DNA damage in mice. Toxicol Sci 2003;73:416-22.
Morris R. Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods 1984;11:47-60.
Kawaguchi S, Kuwahara R, Kohara Y, Uchida Y, Oku Y, Yamashita K. Perinatal exposure to low-dose nonylphenol specifically improves spatial learning and memory in male rat offspring. Indian J Physiol Pharmacol 2015;59:211-22.
Itoh J, Nabeshima T, Kameyama T. Utility of an elevated plus-maze for the evaluation of memory in mice: effects of nootropics, scopolamine and electroconvulsive shock. J Psychopharmacology 1990;101:27-33.
Komada M, Takao K, Miyakawa T: Elevated plus maze for mice. J Vis Exp 2008;22.
Petullo D, Masonic K, Lincoln C, Wibberley L, Teliska M, Yao D. Model development and behavioral assessment of focal cerebral ischemia in rats. Life Sciences 1999;64:1099-108.
Schaar KL, Brennema MM, Savitz SI. Functional assessments in the rodent stroke model. Exp Transl Stroke Med 2010;2:13.
Varlinskaya EI, Spear LP. Social interactions in adolescent and adult Sprague-Dawley rats: impact of social deprivation and test context familiarity. Behav Brain Res 2008;188:398-405.
Kaidanovich-Beilin O, Lipina T, Vukobradovic I, Roder J, Woodgett JRJ. Assessment of social interaction behaviors. J Vis Exp 2011;48:pii: 2473.
Vogel H. Drug discovery and evaluation: pharmacological assays. Springer-Verlag Berlin Heidelberg, Germany (2007).
Wang W, Rusin O, Xu X, Kim KK, Escobedo JO, Fakayode SO, Fletcher KA, Lowry M, Schowalter CM, Lawrence CM. Detection of homocysteine and cysteine. J Am Chem Soc 2005;127:15949-58.
Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal Biochem 1982;126:131-8.
Ellman GL, Courtney KD, Andres V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961;7:88-95.
Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959;82:70-7.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
Grisham MB, Specian RD, Zimmerman TE. Effects of nitric oxide synthase inhibition on the pathophysiology observed in a model of chronic granulomatous colitis. J Pharmacol Exp Ther 1994;271:1114-21.
Patriarca P, Dri P, Snidero MJ. Interference of myeloperoxidase with the estimation of superoxide dismutase activity. J Lab Clin Med 1977;90:289-94.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
Tapia-Rojas C, Lindsay CB, Montecinos-Oliva C, Arrazola MS, Retamales RM, Bunout D, Hirsch Sand Inestrosa NC. Is L-methionine a trigger factor for Alzheimer’s-like neurodegeneration?: changes in Aβ oligomers, tau phosphorylation, synaptic proteins, Wnt signaling and behavioral impairment in wild-type mice. Mol Neurodegener 2015;10:62.
Lehotský J, Tothová B, Kovalská M, Dobrota D, Beňová A, Kalenská D, Kaplán P. Role of homocysteine in the ischemic stroke and development of ischemic tolerance. Front Neurosci 2016l;10:538.
Janaky R, Varga V, Hermann A, Saransaari P, Oja SS. Mechanisms of L-cysteine neurotoxicity. Neurochem Res 2000;25:1397-1405.
Lewerenz J, Maher P. Chronic glutamate toxicity in neurodegenerative diseases-what is the evidence? Front Neurosci 2015;9:469.
Sultana R, Perluigi M, Butterfield DA. Lipid peroxidation triggers neurodegeneration: a redox proteomics view into the Alzheimer disease brain. Free Radic Biol Med 2013;62:157-169.
Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state. Nutr J 2015;15:71.
Pérez-Hernández J, Zaldívar-Machorro VJ, Villanueva-Porras D, Vega-Ávila E, Chavarría A. A potential alternative against neurodegenerative diseases: Phytodrugs. Oxid Med Cell Longev 2016;2016.
Lashley T, Schott JM, Weston P, Murray CE, Wellington H, Keshavan A, Foti SC, Foiani M, Toombs J, Rohrer JD, Heslegrave A, Zetterberg H. Molecular biomarkers of Alzheimer’s disease: progress and prospects. Dis Model Mech 2018;11:031781.
Snowden S, Ebshiana A, Hye A, Yang A, Pletnikova O, Brien O.R, Troncoso J, Legido-Quigley C, Thambisetty M. Neurotransmitter imbalance in the brain and Alzheimer’s pathology. Biorxiv 2017.
Tavakkoli A, Ahmadi A, Razavi BM, Hosseinzadeh H. Black Seed (Nigella sativa
) and its constituent thymoquinone as an antidote or a protective agent against natural or chemical toxicities. Iran J Pharm Res 2017;16:2-23.
Demir E, Taysi S, Ulusal H, Kaplan DS, Cinar K, Tarakcioglu M. Nigella sativa
oil and thymoquinone reduce oxidative stress in the brain tissue of rats exposed to total head irradiation. Int J Radiat Biol 2020;96:228-35.
Lane RM, Potkin SG, Enz A. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int J Neuropsychopharmacol 2006;9:101-24.
Darakhshan A, Pour AB, Colagar AH, Sisakhtnezhad S. Thymoquinone and its therapeutic potentials. Pharmacol Res 2015;95:138-58.
Fattah LIA, Zickri MB, Aal LA, Heikal O, Osama E. The effect of thymoquinone, α7 receptor agonist and α7 receptor allosteric modulator on the cerebral cortex in experimentally induced Alzheimer’s disease in relation to MSCs activation. Int J Stem Cells 2016;9:230.
[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2]