1Hygia Institute of Pharmaceutical Education & Research, Lucknow.
2Ambekeshwar Group of Institutions, Lucknow.
3Deen Dayal University, Gorakhpur.
4Hygia Institute of Pharmaceutical Education & Research, Lucknow.
Introduction: Despite being a chemotherapeutic agent for increasing cancer survival and improving quality of life, Cyclophosphamide (CYP) is unfortunately has left with many side effects such as hepatotoxicity, nephrotoxicity, cardiotoxicity, and cognitive impairment by slow processing speed, recollection impairment, incapability to concentrate, and language problems associated with cognitive impairment in cancer survivors whose exact mechanism is indefinite. Objective: To investigate the protective effect of fluvoxamine, an anti-depressant drug, possessing protection against oxidative stress was used against cyclophosphamide induced cognitive impairment. Thus, to understand this, a clinically relevant dose to cancer treatment was used in mice to establish the chemo brain models, and the spatial memory using multiple behavior tests. Method: 25 mice, weighted 18-25 g were divided 5 animals in each group: Control, disease control (CYP 100mg/kg; i.p; alternate days), standard (donepezil 1mg/kg; orally) and test (fluvoxamine; 100 & 200mg/kg) for 15 days. The mice’s memory was tested using Morris’s water maze, elevated plus maze and modified passive avoidance tests for 3 days, 24 hr after the last dose of CYP. Biochemical parameters like LPO, GSH and SOD were also assessed. Result: The mice treated with fluvoxamine exhibited improvement in memory function in all the behavioral test studies, in Morris’s water maze the test drug at 200 mg / kg showed reduction in transfer latency and increase step down latency level and showed increase in GSH and SOD levels but significantly decreased LPO as compared to cyclophosphamide. Thus, this CICI was significantly ameliorated by the prior administration of fluvoxamine at high dose (200mg/kg). Conclusion: The results suggested that fluvoxamine at a dose of 100; 200mg/kg improves cognitive impairment and brain oxidative damage induced by CYP. Therefore, fluvoxamine could be used as adjuvant chemotherapy to reduce the memory impairments and oxidative stress.
Acquisition, consolidation, and retrieval are the three essential phases of memory, which is a crucial brain function. Acquiring a task is called acquisition; consolidating information stabilizes memory is called retrieval; and bringing the learnt task back is called retrieval.[1] There cannot be recollection in the future without learning.[2] Cognition, which includes a wide range of processes like attention, perception, memory coding, retention, recall, and decision-making along with reasoning, problem-solving, imaging, planning, and action execution, is a part of memory that is known through perception, reasoning, intuition, and knowledge.[1] Farahmandfar M, [2] Johansen JP, [3] Pawlik P, Neuronal networks in the brain use electrochemical signalling for learning and memory functions. A link at the neuronal synapse that may change during learning and memory disruption is known as synaptic plasticity. [1] Chemotherapy is still a useful treatment for a number of different malignancies. Despite the efficacy of medication on improving survival in patients. Chemotherapy is a drug treatment that uses active chemicals in the body to destroy fast-growing cells. Chemotherapy is most broadly used to treat cancer, because cancer cells produce and multiply much faster than most of the body's cells. [4] Skeel R T, Khleuf Chemo fog or cancer treatment brain refers to chemotherapy-induced cognitive impairment or dysfunction, also known as chemotherapy-related cognitive impairment. [5] Mayo Clinic Chemotherapy causes brain damage healthy brain cells to die after long therapy has ended and is one of the biological root causes of chemo brain side effects. Cancer patients have long been complaining about neurobiological dysfunction, such as short-term memory loss, long-term memory loss, epilepsy, lost vision and dementia, depression, stress, anxiety, fear, memory issue, not well understood, trouble finding the right word uncertainty, difficulty concentrating, etc. Related to the diagnosis and treatment of cancer such as parietal zone, temporal frontal and pre-frontal regions affected by cognitive dysfunction caused by chemotherapy. CICI may have a negative effect on long- and short-term quality of life. [6] Meyers C, Cyclophosphamide is the most broadly used anticancer drugs and alkylating agent, with its metabolites causing alkyl cross links between DNA strands of dividing cell causing them to apoptosis. Cyclophosphamide a synthetic broad spectrum anticancer medication and alkylating agent, is an effective therapy for a wide range of cancers including breast, myeloma, ovarian, neuroblastoma and leukemia. It is used to treat chronic and severe leukemia, rheumatoid, lymphomas, and multiple myeloma as well as to prepare for bone marrow transplant. [7] Colvin OM One of the most active sigma-1 receptor agonists is the selective serotonin reuptake fluvoxamine inhibitor. Fluvoxamine is an antidepressant medication, a class of selective serotonin reuptake inhibitor (SSRI) used to treat obsessive compulsive disorder (OCD) and is also used to treat anxiety disorder and depression, such as social anxiety disorder, panic disorder and post traumatic stress disorder. Fluvoxamine worked in long term therapy. It has some analgesic properties with other SSRI and Tricyclic antidepressant (TCA). The exact mechanism of action of fluvoxamine has not been fully determined, but appears to be linked to its inhibition of CNS neuronal uptake of serotonin. Fluvoxamine blocks serotonin reuptake at the neuronal membrane's serotonin reuptake pump, further enhancing serotonin action on 5HT auto receptors. [8] Spina E The purpose of this study is to identify the function of fluvoxamine in mitigating the memory impairment that CYP therapy causes in mice. The purpose of this work is to create a chemo brain model in mice by treating them with CYP, which will lead to cognitive impairment. Then, the study will look at whether fluvoxamine may reverse the memory impairment that the CYP therapy caused. Behavioral tests, such as the Morris water maze, the passive avoidance test, and the elevated plus maze (EPM) tests, are used to assess the cognitive impairment of the model.
2.MATERIAL AND METHODS
2.1Experimental animals
The study utilized Swiss albino mice, of both genders, measuring 20-25g, with consent from the Institutional Animal Ethics Committee. Each mouse was kept in an own housing unit with free access to food and water, and they were kept on a 12-hour light/dark cycle. Every day, the animals were observed and their body weights were recorded. Every behavioral test was run during the cycle's light phase. Prior to behavioral testing, the animals were allowed to adjust to the laboratory environment for a minimum of seven days.
2.2 Drugs and Chemicals
The fluvoxamine tablet was purchased from Solvay pharmaceutical, India. Donepezil (as a gift sample from Vasudha Pharma, Hyderabad), cyclophosphamide (Biochem pharmaceutical industries), Thio barbituric acid, Trichloroacetic acid, Tris HCL buffer, Pyrogallol, Phosphate buffer, Hydrogen peroxide, Ellman reagent. All other solvents were of analytical grade and distilled water was used throughout the study.
2.3 Grouping and treatment protocol
After the acclimatization period, the mice were grouped into 5 groups (n = 5) namely, group 1: vehicle control, group 2: CYP (NC), group 3: donepezil (standard drug), group 4: CYP + fluvoxamine (low dose), and group 5: CYP + fluvoxamine (high dose). The mice of group 2, 4 and 5 were intraperitoneally (i.p.) injected with CYP (1mg/kg). Group 1 was provided purified drinking water throughout the study. Donepezil (1 mg/ml) was admixed in the drinking water and given from a day before CYP treatment during the entire study period. After the animals received CYP doses, they were subjected to behavioral tests. All the behavioral tests were performed during the light phase of the cycle with uniform lighting conditions (30 ± 2 lux). The animal wellbeing and body weight were monitored daily. [9] Jyoti G,
Table No. 1 Grouping and treatment protocol
|
Group |
Treatment plan |
Route |
No. of animals |
|
Group 1 |
Positive control |
Normal saline |
5 |
|
Group 2 |
Negative control (cyclophosphamide 1mg/kg) |
i.p |
5 |
|
Group 3 |
Standard drug (donepezil 1mg/kg) +CYP |
i.p |
5 |
|
Group 4 |
Low dose of fluvoxamine (100mg/kg) +CYP |
p.o |
5 |
|
Group 5 |
High dose of fluvoxamine (200mg/kg) +CYP |
p.o |
5 |
2.4 Acute toxicity study
Prior to administering the test chemical, mice were allowed to fast for the entire night and were given only 3-5 ounces of water. The oral dosage of fluvoxamine was increased to 50, 100, 500, 1000, 2000, and 4000 mg/kg of body weight. The mice were given a dose, and within four hours they were monitored for signs of toxicity, including tremors, convulsions, muscle spasm, tonic extension, loss of the righting reflex, ataxia, drowsiness, hypnosis, lacrimation, diarrhea, salivation, and writhing during motor activity. After then, the mice were monitored for up to 72 hours to check for death.
3. Memory models
3.1 Induction of cognitive impairment
Cognitive impairment was induced in mice by using donepezil at a dose of 1 mg/kg, i.p only once during the model. It was given 30 min before retention trial in MWM, and 30 min before acquisition trial in EPM. In modified passive avoidance, it was injected to animals 30 min before acquisition trial
3.2 Morris water maze
A white circular pool of 45 cm in diameter and 26 cm in height, with water at 20°C ± 1°C to a depth of 30 cm, served as the maze. The animals received the therapy for a period of 21 days. On different days, cyclophosphamide injections were given. Starting on day 13, the animals were trained four times a day, with a five-minute interval between trials, for the maze task.
Mice that were unable to locate the platform in the 120-second window (the first learning period is 180 seconds) were positioned to the side and given a 30-second window to remain there. The animals were examined three times, with each trial taking place between thirty and forty-five minutes after the injection of cyclophosphamide in water that had been rendered opaque by the addition of milk. The number of crossings, residence time, and escape latency were assessed as testing factors for spatial learning. [10,11,12 ] (Kiran et al.,2012; Rajasekharet al.,2013; Alikatteet al.,2012)
3.3 Elevated Plus Maze
The EPM test is a behavioral test that is commonly used to measure the learning and memory processes. It consisted of two opposing arms. The open arm’s length was 30 cm and width was 5 cm, while the closed arm’s length was 30 cm and width was 5 cm. The height of the sidewalls was 15 cm. An open central area measured 5 cm2, and the maze was elevated 30 cm above the floor. During the acquisition trial, each mouse was individually placed at the end of the open arm, facing opposite the central platform. The mice were allowed to explore the apparatus for 5 min. After 3 h, each mouse was placed facing opposite the central platform. The latency time (LT) was recorded. The LT is the time it took a mouse to move from the end of the open arm and place all four of its paws inside either of the closed arms. [13, 14] Nigam A, Komada M,
3.4 Modified passive avoidance
The passive avoidance apparatus uses negative reinforcement to evaluate long-term memory. The apparatus was made out of a 20 cm × 20 cm × 20 cm box with three wooden walls and one Plexiglas wall. The grid floor was made of 3 mm stainless steel rods, and in the center of the grid floor was a wooden platform measuring 10 cm × 7 cm × 1.7 cm. Throughout the trial, a 15 W bulb was used to light the box. The animal received each treatment for a period of 21 days. Following a 30-minute injection of cyclophosphamide, each mouse was positioned on a wooden platform situated in the middle of the grid floor. Step-down latency (SDL) was recorded when the mouse stepped down and placed its paw on the grid floor. Foot shock (50 Hz; 1.5 mA; 1 s) was then applied. The amount of time it takes the mice to step down and plant all four paws on the grid floor is known as the SDL. For the acquisition and retention tasks, mice with SDL in the 2–15 s range were used. [ 15,16,17]
3.5 Biochemical estimation of markers of oxidative stress
Mice were slaughtered by cervical dislocation on the fifteenth day following the conclusion of the behavioral trials. Each rat's brain was removed and stored on an ice-cold platter.
3.6 Preparation of brain homogenate
To evaluate MDA, GSH, and AChE activity, the brains were homogenized ten times using ice-cold 0.1 M phosphate buffer (pH 7.4). [18]. Tota S,
3.7 Estimation of acetylcholinesterase’s activity
The AChE activity was measured by the method of ellman et al. (1961) with slight modification. Change in absorbance per minute of the sample was read spectrophotometrically at 420nm.
3.8 Measurement of glutathione level
By detecting the yellow chromophore that emerged from GSH's interaction with DTNB (Ellman reagent), whose absorbance was ascertained via spectrophotometry, the amount of GSH was ascertained. The brain homogenate was mixed with a 10% trichloroacetic acid (TCA) solution, and the combination was centrifuged for 10 minutes at 4°C and 200 rpm. The supernatant was used to quantify GSH. Add 2 ml of phosphate buffer (pH 8.4), 0.4 ml of double-distilled water, 0.5 ml of DTNB, and 0.1 ml of processed tissue sample. Gently tremble in a vortex. To read the absorbance at 412 nm, it took 15 minutes. The GSH level was expressed as nmol/mg protein. [19] Jawaid T,
3.9 Measurement of malondialdehyde level
Following a thorough mixing process, the mixture was centrifuged at 3,000 g for 10 minutes after adding 0.5 ml of tissue homogenate, 0.5 ml of distilled water, and 1.0 ml of 10% TCA. To 0.2 ml of supernatant, 0.1 ml TBA was added. The entire combination was allowed to cool to room temperature after 40 minutes at 80°C in a water bath. The absorbance of the clear supernatant was measured at 532 nm using a spectrophotometer. The MDA protein level was expressed as nmol/mg.[19,20]
3.10 Estimation of Superoxidase dismutase (SOD)
The elucidation of riboflavin solution in the existence of EDTA reduces the Flavin. It then adds oxygen and lowers oxygen to O-2, a fragment of the detector which allows the NBT to react, lowers the NBT to formazan blue. The sample SOD impedes the manufacture of formazan. For a total volume of 0.2ml of 0.1 M EDTA and 0.1ml of 1.5 mM NBT and a phosphate buffer of 2.6 ml, the absorbance of 0.01ml of homogeneous mixed at 560 nm was measured. Both tubes held 15 minutes in the incubator, and blue color absorption has been tested again. The percentage of the reserve had been considered after comparing sample absorbance and control absorbance.[21] DURAK I,
|
Treatment groups mg/kg |
Acquisition latency (sec) Before Cyclophosphamide |
Retention latency (sec) After Cyclophosphamide |
|
|
Day 13 |
Day 14 |
|
|
|
Positive Control |
38.40 ± 2.66 |
24.80 ± 2.69 |
Positive Control
|
|
Negative Control |
40.40 ± 1.91 |
35.60 ± 2.37 |
Negative Control
|
|
Standard |
36.60 ± 1.42 |
30.40 ± 1.02 |
Standard |
|
Test 1 |
28.80 ± 3.41 |
25.40 ± 1.47 |
Test 1 |
|
Test 2 |
33.40 ± 1.94 |
23.80 ± 1.68 |
Test 2 |
3.11 Statistical Analysis
Statistical analysis was performed by one-way analysis of variance (ANOVA) with Tukey’s post-hoctest. Values are expressed as mean ± standard error of the mean (SEM).
4. RESULTS AND DISCUSSION
4.1 Acute Toxicity Studies
fluvoxamine did not cause any mouse death in the acute toxicity trial. While 200 and 400 mg/kg doses were used to assess various activities, even at this higher dose of 4000 mg/kg, no gross behavioral changes were observed, including skin, fur condition, eyes color, motor activity, convulsions, tonic extension, loss of righting reflex, ataxia, sedation, hypnosis, and writhing.
4.2 Effect of drugs on cyclophosphamide ?induce memory impairment in Morris’s water maze test
In Morris water maze test cyclophosphamide (100 mg/kg) increased the latency to recognize the hidden platform and the cyclophosphamide induced impairment was signi?cantly ameliorated by the prior administration of fluvoxamine at a dose of 100 mg/kg. Donepezil (1mg/kg i.p.) showed the maximum reduction (Table 3.4 and fig. 3.5) indicating an e?ect on progress in learning and memory in retention trial. Though, cyclophosphamide increased the latency to find hidden platform, but at the end of experiment (15th day), in fluvoxamine treated groups (100 & 200 mg/kg), escape latency was (24.80 ± 1.11***) s and (21.20 ± 1.96***) s respectively. Donepezil (1 mg/kg) showed maximum reduction (20.2 ± 0.94***) in escape latency.
Table No.2 Effect of fluvoxamine on Escape Latency in cyclophosphamide induced memory impairment in mice.
Results expressed as Mean ± SEM (n=5) and ***P<0.0001 as Compared with negative control group by One Way ANOVA followed by Tukey Test.
There was a difference in the mean EL of cyclophosphamide treated groups compared to test groups. The test drug fluvoxamine at a dose of 200 mg showed decrease in TL when compared to the negative control group.
Fig. No. 1 Effect of fluvoxamine on Escape Latency in cyclophosphamide induced memory impairment on mice Day 15.
Results as P value are Mean ± SEM (n=5) and P<0.0001*** as Compared with negative control group by One Way ANOVA by Tukey Test
4.3 Number of crossing and Residence Time
Two other parameters i.e numbers of crossings and residence time in target quadrants in animals after 14 days treatment with fluvoxamine and donepezil shows considerable increase when compared to negative control (p < 0.0001) and control group. At the end of experiment (15th day), the number of crossings of the fluvoxamine dosed groups at the dose of (100 and 200 mg/kg) were (5.80 ± 0.40**) and (6.00 ± 0.33**) and residence time in target quadrant was (5.60 ± 0.40) and (8.00 ± 0.49**) respectively. Donepezil (1 mg/kg) showed maximum increase in number of crossings (7.00 ± 0.46***). As well as increases in residence time (9.60 ± 0.60***).
Table No. 3 Effects of Fluvoxamine on No. of crossings and Residence time in cyclophosphamide induced memory impairment in mice
|
S. No. |
Treatment |
No. of crossings |
Residence Time |
|
1 |
Positive Control |
8.60 ± 0.53 |
10.60 ± 0.51
|
|
2 |
Negative Control |
3.40 ± 0.19 |
4.80 ± 0.40 |
|
3 |
Standard |
7.00 ± 0.46*** |
9.60 ± 0.60***
|
|
4 |
Test 1 |
5.80 ± 0.40** |
5.60 ± 0.40 |
|
5 |
Test 2 |
6.00 ± 0.33** |
8.00 ± 0.49**
|
Results expressed as Mean ± SEM (n=6) by One way ANOVA followed by Tucky test
Fig No. 2 Effects of Fluvoxamine on Number of crossings by cyclophosphamide induced memory impairment in mice
Fig. No. 3 Effects of fluvoxamine on time spent in target quadrant by cyclophosphamide induced memory impairment in mice.
4.4 Effect of drugs on cyclophosphamide ?induce memory impairment in Elevated plus maze test
The effect of treatment on negative control, test group and standard group (1 mg/kg) were evaluated at end of the day 15. The cyclophosphamide (100 mg/kg) treated rodents showed increases in TL times ??on days of acquisition as well as retention, learning and memory process. On day 15, fluvoxamine at a dose level of 200 mg / kg established a significant reduced in transfer latency compared to the cyclophosphamide received animal. Donepezil (1 mg / kg) demonstrated a significant reduction in TL compared to the cyclophosphamide animal.
Table No. 4 Effect of Fluvoxamine on transfer latency in cyclophosphamide induced memory impairment in mice
|
Treatment Group |
Transfer Latency |
|
|
|
Acquisition Day 14 |
Retention day 15 |
|
Positive Control |
22.60 ± 1.15 |
14.00 ± 0.83 |
|
Negative Control |
21.40 ± 0.83 |
24.60 ± 1.65 |
|
Standard |
19.20 ± 0.78 |
17.00 ± 0.58*** |
|
Test 1 |
23.40 ± 0.88 |
18.80 ± 0.59** |
|
Test 2 |
20.80 ± 1.35 |
15.80 ± 0.78*** |
Results as P value are Mean ± SEM (n=5) and P< 0.001**, P< 0.0001*** as Compared with negative control group by One Way ANOVA by Tukey Test.
In the present experiment, mean TL on day 14 for each rat was relatively stable and showed no significant variation among different groups. following training, the test drug treated (100, 200mg/kg) rats entered closed arm quickly as compared negative control group rats. Mean transfer latency on 15 days were shorter as compared to TL on 14 days in standard & test drugs respectively.
Fig No 4 Effects of fluvoxamine on Transfer latency in cyclophosphamide induced memory impairment mice on Day 14
Fig No. 5 Effects of fluvoxamine on Transfer latency in cyclophosphamide induced memory impairment mice on Day 15
4.5 Effect of drugs on cyclophosphamide ?induce memory impairment in Modified Passive Avoidance
Cyclophosphamide treatment (100 mg / kg i.p) caused a decrease in step down latency reduces indicating memory impairment. An increase in fluvoxamine latency level at a dose of (100, 200 mg / kg) and reversed cyclophosphamide-induced memory impairment. The group of rodents dose with donepezil (1 mg / kg, i.p) showed significant memory improvement and reversed the memory impairment induced by cyclophosphamide.
Table No. 5 Effect of drugs on cyclophosphamide ?induce memory impairment in Modified Passive Avoidance
|
Treatment Group |
Step Down Latency |
|
|
|
Step Down Latency (Day 1) |
Step Down Latency (Day 2) |
|
Positive Control |
16.40 ± 0.81 |
23.40 ± 1.55 |
|
Negative Control
|
14.40± 1.13 |
9.60 ± 0.51 |
|
Standard
|
11.80 ± 1.02 |
15.40 ± 0.92** |
|
Test 1
|
10.60 ± 0.64* |
12.20 ± 0.81 |
|
Test 2
|
8.80 ± 0.43** |
10.80 ± 0.94 |
Results as P value are Mean ± SEM (n=5) and P<0.001*, P<0.001** as Compared with negative control group by One Way ANOVA by Tukey test
Fig. No.6 Effects of Fluvoxamine on Step down latency in cyclophosphamide induced memory impairment in mice on Day 1
Fig. No.7 Effects of Fluvoxamine on Step down latency in cyclophosphamide induced memory impairment in mice on Day 2
4.6 Biochemical parameter:
4.6.1 Effect of drugs on glutathione level
Glutathione was estimated in brain after the completion of behavioral studies. A significant decrease in the level of GSH was observed in the negative control group as compared to the positive control group. There was a significant rise level of GSH in brains of treated with high dose of fluvoxamine in comparison to cyclophosphamide group. Standard group showed increase in GSH level in cyclophosphamide?treated animals.
4.6.2 Effect of drugs on MDA level
The MDA level was estimated in rat brain after the completion of behavioral studies. MDA level increased significantly (P < 0.001) in the brain of cyclophosphamide?treated rats as compared to the positive control group. On the other hand, donepezil significantly decreased (P < 0.001) the MDA level in comparison to cyclophosphamide group. Fluvoxamine dose at of 200 mg/kg, p. o. showed significant decline in brain MDA level when compared to negative control group.
4.6.3 Effect of drugs on SOD level
Cyclophosphamide treatment increased brain SOD activity in compare to vehicle control group. When compared to the cyclophosphamide treated group, donepezil and fluvoxamine (100 and 200 mg/kg, p.o) treatment decline in the SOD activity. Fluvoxamine at a dose of 100 mg/ kg and 200mg/kg p.o. showed decrease in brain SOD level in a dose dependent manner when compared to standard drug treatment.
Table No. 6 Effect of drugs on glutathione, MDA and SOD level
|
Treatment groups |
MDA |
GSH |
SOD |
|
Positive Control |
7.43 ± 0.09
|
0.97 ± 0.05
|
1.86 ± 0.37
|
|
Negative Control
|
10.18± 0.13 |
0.08 ± 0.01 |
0.57 ± 0.04
|
|
Standard
|
8.23 ± 0.09***
|
0.62 ± 0.04 |
1.45 ± 0.31*** |
|
Test 1
|
9.83± 0.04 |
0.12 ± 0.01 |
0.63 ± 0.02 |
|
Test 2
|
9.66 ± 0.05** |
0.37 ± 0.04 |
1.24 ± 0.07*** |
Result expressed as Mean ± SEM (n=6) and **P<0.001, ***P<0.0001 as Compared with negative control group by One Way ANOVA by Tukey Test.
Fig.No. 8 Effect of fluvoxamine on SOD activity of cyclophosphamide induced memory impairment
Fig. No. 9 Effect of fluvoxamine on MDA activity of cyclophosphamide induced memory impairment
Fig. No. 10 Effect of fluvoxamine on GSH activity of cyclophosphamide induced memory impairment
5 DISCUSSIONS
Many chemotherapeutic agents could directly or indirectly affect brain function [26,27]. Cyclophosphamide is one of the most widely used chemotherapeutic agents for breast cancer patients [5]. The active mechanism of the cyclophosphamide binds to the DNA and prevents replication of DNA, inducing apoptosis. However, chronic CYP administration can cause toxicity to other non-target tissues, leading to adverse effects such as alopecia, nausea, fatigue and cognitive impairment.
This present study tested the protective effects of fluvoxamine on cyclophosphamide-induced memory impairment by chemobrain mouse model, i.e. the Morris water maze (MWM), elevated plus maze (EPM), passive avoidance test.
The results of this study showed that the memory impairment occurred due to CYP treatment, and these impairments were rescued through the administration of fluvoxamine assessed by behavioral tests and biochemical tests.
The studies had showed that intra-peritoneal administered cyclophosphamide caused substantial oxidative stress in the central nervous system. It occurs as a result of increased levels of malondialdehyde in the brain and lipid peroxidation. Cyclophosphamide also inhibits brain, heart and lung catalase and superoxide dismutase antioxidant capacity. Cyclophosphamide also decreases levels of glutathione.
In multiple experimental models, Sigma-1 receptor agonists show important anti-amnestic and neuro-protective effects and fluvoxamine has been reported to be clinically successful in some cognitive impairment-related psychopathological conditions.
In the present dissertation, the results of the MWMz test in all groups throughout the training stage show a normal learning profile, as indicated by a decrease in migration conditions. In the Morris water maze trial, cyclophosphamide treatment increased the hidden platform to and cyclophosphamide-induced impairment by prior use of fluvoxamine at doses of 100 to 200 mg / kg. Donepezil (1mg / kg p.o) showed a maximum decrease in retention testing indicating improvement in learning and memory.
Elevated plus-maze is used to assess the animal’s learned behavior. However, transfer latency was markedly reduced if the rodents had previously learned in entering to the arms of EPM and shorter transfer latency has been related to the memory process. In the present study fluvoxamine orally enhanced the learning and memory of mice.
The effect of treatment on negative control, test group and standard group (1 mg/kg) were checked at the end of day 14. The cyclophosphamide (100 mg/kg) treated group showed a significant increase in TL values on the acquisition as well as on the retention days, indicating impairment in learning and memory. On the day 15, the Fluvoxamine at the dose level of 100 and 200 mg/kg and donepezil showed a significant decrease in the TL as difference to the cyclophosphamide control group.
Passive avoidance activities are based on negative support and is used to examine long-term memory. Acquisition of modified passive avoidance cyclophosphamide (100 mg / kg i.P) decreased step by step in the retention test indicating latency and memory impairment. At a dose of (100, 200 mg / kg) fluvoxamine increases step-down latency and reverses cyclophosphamide-induced memory impairment. The group of mice that were treated with donepezil (1 mg / kg, p.o.) showed significant improvement in memory and reversed memory impairment induced by cyclophosphamide. Oxidative stress and antioxidant systems play important roles in pathological neurological changes.
Antioxidants are essential for good health. Antioxidants can protect the body from these potentially harmful substances. Antioxidants are agents that fight the harmful process of oxidation, which afflicts the integrity of the cell. Cells are damaged or oxidized because they attacked with free radicals. Free radicals are molecules with a free electron in their outer shell that makes them highly reactive with other cells. When a free radical comes into contact with another cell, it will attack, oxidize and damage it. Oxidative stress is one of the earliest events in the pathogenesis of memory loss. Lipid peroxidation is a well-known element that plays an essential role in oxidative stress balance. Oxidative stress affects both short-term memory and long-term memory.
In the present study, cyclophosphamide treatment reduced brain glutathione activity compared to the positive control group. Donepezil and fluvoxamine (100 and 200mg / kg, p. O) treatment increased glutathione activity compared to the cyclophosphamide treated group. Fluvoxamine at a dose of 200mg / kg, p.o showed a significant increase in brain glutathione levels compared to the negative control group. In this study cyclophosphamide induced increased the MDA level when compared to positive control mice. Donepezil appreciably reduced (p<0.001) MDA amount compared to mice provided with cyclophosphamide. The administration of fluvoxamine (100mg / kg) did not demonstrate any significant increase in the amount of MDA compared to cyclophosphamide-administered mice. Administration of fluvoxamine at a dose of 200 mg / kg, MDA decreased dramatically as compared to the negative control group.
The action of SOD is a susceptible indicator in oxidative disrupt because it scavenges superoxide ions to produce hydrogen peroxides to reduce noxious effects. Cyclophosphamide treatment increased brain SOD activity compared to the positive control group.
Donepezil & fluvoxamine treatment decreased the SOD activity when compared to the cyclophosphamide treated group. Fluvoxamine at a dose of 100mg/ kg and 200mg/kg p.o. showed decrease in brain SOD level in a dose dependent manner when compared to standard drug treatment.
6. CONCLUSION-
Despite efforts to improve chemotherapeutics cancer treatment over the years the survival rate and quality of life in advanced cancer have increased however cancer survivor are left with many side effects such as hepatotoxicity, nephrotoxicity, cardio toxicity, and cognitive impairment.
Cancer survivor has slow processing speed, recollection impairment, incapability to concentrate, and language problems, is called chemotherapy induced cognitive impairment.
The results from the present study suggested fluvoxamine at a dose of 100,200mg/kg improves cognitive impairment and brain oxidative damage induced by cyclophosphamide using chemo brain behavioral models treated with cyclophosphamide. The result of this study revealed that memory impairment occurred due to cyclophosphamide treatments and these impairments were rescued through the administration of fluvoxamine. Therefore, fluvoxamine could be used as a adjuvant chemotherapy to reduce the memory impairments and oxidative stress.
Oxidative stress is one recognized mechanism underlying chemotherapy induced memory impairment connected with cyclophosphamide (CYP) treatment. An enhance in oxidative stress or decline in the antioxidant ability of the brain is input factor concerned in neuronal degeneration in the elderly.
ACKNOWLEDGEMENTS
The authors are grateful to the hygia Institute of Pharmaceutical Education and Research, Lucknow, India, for providing necessary facilities to carry out this research. The authors would also like to thank the Central Drug Research Institute, Lucknow, for providing animals. The study was supported by a ?nancial grant to T. J. from the hygia Institute of Pharmaceutical Education and Research. The funding agency had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
REFERENCES
Chandani Nigam*, Dr. Dharamveer Panjwani, Ashirvad Jaiswal, Sakshi Srivastava, Investigation of the Protective Effect of Fluvoxamine on Cyclophosphamide Induced Cognitive Impairment in Laboratory Animals, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 12, 939-953 https://doi.org/10.5281/zenodo.17830602
10.5281/zenodo.17830602
