TO DETERMINE THE ROLE OF NARIGENIN IN ANXITY MODELS OF SWISS ALBINO MICE

Abstract

ABSTRACT The present invention relates to evaluating the pharmacological potential of Naringenin, a flavonoid compound, in the treatment of anxiety disorders. The study was conducted using Swiss albino mice models to assess both in-vitro antioxidant activity and in-vivo sedative and anxiolytic effects. The antioxidant activity was measured using the Nitric Oxide Radical Scavenging method, showing significant free radical inhibition by the methanol and petroleum ether extracts of Naringenin. The in-vivo sedative and anxiolytic activities were tested using the Elevated Plus Maze and Astrophotometer methods. Naringenin at doses of 50mg/kg and 100mg/kg demonstrated significant anxiolytic effects by increasing open arm entries and reducing locomotor activity. Results indicate that Naringenin has a potential therapeutic role in managing anxiety disorders by enhancing GABA receptor activity. This study opens avenues for developing Naringenin-based anxiolytic therapies with fewer side effects than conventional benzodiazepines.

Specification

Description:FIELD OF INVENTION
The present invention relates to field of pharmacology, particularly to the use of natural compounds in managing mental health disorders. Specifically, this invention focuses on the pharmacological effects of Naringenin, a flavonoid, in reducing anxiety and promoting sedation as demonstrated in Swiss Albino mice.

BACKGROUND OF THE INVENTION
Anxiety disorders are among the most prevalent mental health conditions globally, significantly affecting individuals' daily functioning and quality of life. Current treatments, including benzodiazepines, selective serotonin reuptake inhibitors (SSRIs), and serotonin-norepinephrine reuptake inhibitors (SNRIs), have proven effective but are often accompanied by severe drawbacks such as dependency, tolerance, delayed onset of action, and adverse side effects, including drowsiness, cognitive impairment, and gastrointestinal disturbances. Furthermore, these medications fail to address oxidative stress, a critical contributor to the pathophysiology of anxiety disorders, characterized by an imbalance between reactive oxygen species and the body's antioxidant defense mechanisms. The high cost and limited accessibility of conventional anti-anxiety drugs, especially in resource-constrained settings, further exacerbate the challenge of managing anxiety disorders effectively. This invention addresses these shortcomings by utilizing Naringenin, a natural flavonoid with demonstrated antioxidant properties and the ability to modulate GABA receptor activity. By targeting both oxidative stress and GABAergic pathways, Naringenin offers a safe, effective, and affordable alternative to conventional anxiety treatments, with reduced risks of dependency and side effects.
OBJECTS OF THE INVENTION
Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows.
It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative
An object of the present disclosure is to reduce reliance on synthetic anxiolytic drugs. Synthetic drugs like diazepam, while effective, often lead to adverse effects such as dependency, drowsiness, and cognitive impairment. Naringenin offers a natural, non-addictive alternative with comparable efficacy, reducing the risk of long-term side effects.
An object of the present disclosure is to exploit Naringenin's dual antioxidant and anxiolytic properties. Oxidative stress is a major contributor to mental health disorders, and Naringenin's strong antioxidant activity addresses this directly by neutralizing free radicals, thereby protecting neural tissues and promoting mental well-being.
An object of the present disclosure is to establish a cost-effective therapeutic alternative. Naringenin is derived from readily available sources like citrus fruits, making its production economical. This can significantly reduce the cost of treatment for anxiety-related disorders, making it accessible for wider populations.
An object of the present disclosure is to validate the use of traditional plant-based remedies through scientific methods. This research bridges traditional medicine and modern pharmacology by scientifically validating the anxiolytic potential of Naringenin, promoting its integration into mainstream therapeutic practices.
An object of the present disclosure is to minimize side effects associated with anxiety treatments. The study's findings show that Naringenin, even at higher doses, exhibits minimal side effects compared to synthetic alternatives. This enhances patient compliance and improves overall treatment outcomes.
An object of the present disclosure is to offer a natural, non-addictive alternative to benzodiazepines for the treatment of anxiety disorders. Naringenin, derived from natural sources, significantly reduces anxiety-like behaviors in animal models without inducing dependency or withdrawal symptoms, thereby addressing a critical limitation of conventional anxiolytics.
An object of the present disclosure is to provide a safer therapeutic option for long-term management of anxiety. The use of Naringenin ensures a favorable safety profile, with minimal adverse effects on cognitive functions. This makes it suitable for chronic use, unlike benzodiazepines that are often limited to short-term treatment due to their side effects.
An object of the present disclosure is to provide formulation that can easily integrated into existing therapeutic regimens. It can serve as an adjunct therapy to reduce the dosage of conventional anxiolytics, thereby minimizing their side effects while enhancing overall treatment efficacy.
Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.
SUMMARY OF THE INVENTION
The following presents a simplified summary of the invention in order to provide a basic understanding of some aspects of the invention. This summary is not an extensive overview of the present invention. It is not intended to identify the key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concept of the invention in a simplified form as a prelude to a more detailed description of the invention presented later.
The present invention focuses on the therapeutic use of Naringenin, a naturally occurring flavonoid, for managing anxiety disorders. This study leverages the antioxidant and anxiolytic properties of Naringenin, tested in Swiss albino mice. The in-vitro antioxidant activity was assessed using the Nitric Oxide Radical Scavenging method, demonstrating significant free radical inhibition. In-vivo studies involved evaluating the sedative and anxiolytic effects using the Elevated Plus Maze and Astrophotometer models. Naringenin administered at 50mg/kg and 100mg/kg doses showed a notable reduction in anxiety-like behaviors, comparable to standard diazepam treatment, without inducing sedation. The invention offers a natural, non-addictive alternative to conventional anxiolytics by targeting oxidative stress pathways and enhancing GABA receptor activity. This innovation presents a promising therapeutic option for long-term anxiety management, with a favorable safety profile and minimal side effects.


DETAILED DESCRIPTION OF THE INVENTION
The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the elements described without departing from the scope of the invention.
Example 1 : PHARMACOLOGICAL INVESTIGATION
1) In-vitro antioxidant activity
2) In-vivo sedative and antianxiety activity

ANTIOXIDANT
Antioxidant is any substance that, when present at low concentration compared with those of an oxidizing substrate, significantly prevents or delays the oxidation of that substrate. Neutralizes free radicals and prevents cell damage that may lead to cancer. Any nutrient or chemicals that react with and neutralize free radicals to prevent oxidative damage to cells (e.g., oxidation of lipid membranes, DNA damage). A good biological antioxidant is able to accept an unpaired electron to form a free radical intermediate with a relatively long half-life in the normal biological environment. There is a complex intracellular enzymatic antioxidant system, including superoxide dismutase, catalase and enzyme of the glutathione peroxidase family. Nonenzymatic antioxidant includes arginine, vitamins A, C, E, B carotene, glutathione, polyphenols and minerals (selenium and zinc).69
IN-VITRO ANTIOXIDANT ACTIVITY
Oxidative stress is an imbalance between cellular production of reactive oxygen species and the counteracting antioxidant mechanisms. The brain with its high oxygen consumption and a lipid-rich environment is considered highly susceptible to oxidative stress or redox imbalances. Therefore, the fact that oxidative stress is implicated in several mental disorders including depression, anxiety disorders, schizophrenia and bipolar disorder, is not surprising.70 Some of the studies suggest that oxidative stress causes anxiety-related behaviors but do not explain the neurobiological pathways underlying the effect of oxidative stress on anxiety symptoms. Some articles showed the use of antioxidant in the prevention or reduction of high anxiety.71 This may be due to the reason that GABA receptors activities are enhanced by antioxidants.72-75 the benzodiazepines act by binding to GABA receptor which is used as both sedative and anti anxiety drug predominantly by patients. Anti-oxidants like poly phenols and flavonoids are therefore very helpful in reduction of stress factors and free radical formation which inhibits GABA binding activity. It has become evident that flavonoids are able to exert enhancement ofGABA binding activity even at low concentration.68 Based on this assumption between the anti-oxidant and the anxiety disorder, the antioxidant activity was conducted for all the extracts using the Nitric oxide radical scavenging activity. In vitro antioxidant activity was done for the two extracts (methanol and petroleum ether) using nitric oxide scavenging activity.
NITRIC OXIDE RADICAL SCAVENGING ACTIVITY:
Sodium nitroprusside (SNP) was measured according to the method of Marcocci et al. (1994). Briefly, the reaction mixture (5.0 ml) containing SNP (5 mM) in phosphate buffered saline (pH 7.3), with or without the plant extract at different concentrations, was incubated at 25°C for two hours.The NO. radical thus generated interacted with oxygen to produce the nitrite ion (NO. ) which was assayed at 30 min intervals by mixing 1.0 ml of incubation mixture with an equal amount of Griess reagent (1% sulfanilamide in 5% phosphoric acid and 0.1% naphthylethylenediaminedihydrochloride). The absorbance of the chromophore (purple azo dye) formed during the diazotisation of nitrite ions with sulphanilamide and subsequent coupling with naphthylethylenediamine dihydrochloride wasmeasured at 546 nm. A control was taken without test compound or standard. Ascorbic acid was taken as a reference antioxidant. All the tests were performed in triplicate and the results averaged. The percentage inhibition of nitric oxide generated was measured by comparing the absorbance values of control and test samples using following formula.
% Radical scavenging activity = [(Abs) control - (Abs) sample / (Abs) control)] ×100
Where, (Abs) control is the absorbance of the control, and (Abs) sample is the absorbance of the test compound. The IC50 values (concentration of sample required to scavenge 50% of free radicals) were calculated from the regression equation, prepared from the concentration of the samples and percentage inhibition of free radicals.77-79
IN-VIVO SEDATIVE AND ANTIANXIETY ACTIVITY
Example 2 : Elevated plus maze in mice were used to evaluate antianxiety activity
II Actophotometer/open field was used to evaluate spontaneous locomotor activity
I EVALUATION OF ANTIANXIETY ACTIVITY IN MICE
1) Elevated plus-maze in mice: 21, 62, 80, 81, 83
Swiss albino mice divided into 4 groups each group containing 6 animals. The entire animals were freely accessible to water and food.
Drug administration:
Normal saline as vehicle, diazepam as standard and the test compound were administered orally using orogastric tube, 60 minutes prior to introduction of mice into elevated plus-maze.Group
I: control group received normal saline, 0.5% per orally
Group II: standard group received diazepam, in the dose of 1mg/kg per orally
Group III: Extracts of Naringenin, in the dose of 50mg/kg per orally
Group IV: Extracts of Naringenin, in the dose of 100mg/kg per orally
The antianxiety activity of plant extracts was evaluated using the Elevated plus-maze test. The apparatus consists of two open arms of 18×4cms and two closed arms of 18.4×4×16cms having an open roof, with the plus-maze elevated to 25cms from the floor. Sixty minutes after oral administration of the test and standard drugs, each animal was placed at the center of the maze facing the open arm of the maze. During the 5 min test period, the number of entries and the time spent in open and enclosed arms were recorded. An entry into an arm was defined as the point when the animal places all four paws onto the arm. Following each trial using 70% ethanol thoroughly clean the apparatus. The behavior of the animal scored by using hand operated counters and stop watches. The procedure was carried out in a sound attenuated room; observations were made from an adjacent corner. The percentage of time spent and entries in the open arm were considered as index.
1. Percentage of open arm entries= (number of open arm entries /number of open arm entries+ number of closed arm entries) ×100
2. Percentage time spent in open arms= (time spent in open arm/time spent in open arm+ time spent in closed arm) ×100
II EVALUATION OF LOCOMOTOR ACTIVITY IN MICE
Two well established models were used to evaluate the locomotor activity
1. Actophotometer
2. Open field test in mice
1) Actophotometer: 21, 62
Albino mice were divided into four groups each containing 6 animals. All the animals were freely accessible to food and water.
Drug administration:
Normal saline as vehicle, diazepam as standard and the test compound were administered orally using orogastric tubes
Group I: control group received normal saline, 0.55% per oral
Group II: standard group received diazepam, in the dose of 1mg/kg per oralGroup
III: Extracts of Naringenin, in the dose of 50mg/kg per oral
Group IV: Extracts of Naringenin, in the dose of 100mg/kg per oral
To evaluate locomotor activity, each mice was placed into an actophotometer and its score of locomotor were measured for a period of 10 minutes. The locomotor activity was measured using a digital actophotometer which operates on photoelectric cells which are connected in circuit with a counter. When the beam of light falling on the photocell is cut off by the animal, a count is recorded and displayed digitally. Mice were administered with control, standard and test compounds 1 hour prior to the experiment and the response taken for 30 minutes once for 1 hour and were tested for activity score for 10 minutes. Decreased activity score was taken an index of CNS depression.
2) Open field method 81, 82
Albino mice were divided into 4 groups each containing 6 animals. All the animals were freely accessible to food and water.
Drug administration
Normal saline as vehicle, diazepam as standard and the test compound were administered orally using orogastric tubes.
Group I: control group received normal saline, 0.5% per oral
Group II: standard group received diazepam, in the dose of 1mg/kg per oral
Group III: Extracts of Naringenin, in the dose of 50mg/kg per oral
Group IV: Extracts of Naringenin, in the dose of 100mg/kg per oral
The experiment was carried out to determine depressive action of the test drugs on CNS in mice. Open field apparatus is taken for the experiment which consists of 16 squares, each side of square having the length of 24 cm and height 17 cm. the total length is 97 cm on each side of the apparatus. The number of squares visited by the animals was counted for 10 min at 30 and 60 min after oral administration of the test and standard drugs.
STATISTICAL ANALYSIS84
All the values were expressed as mean ± SEM. The data was statistically analyzed by one way ANOVA followed by Dunnet's test. One way analysis of variance (ANOVA) was used to correlate the statistical difference between the variables. P<0.05 was considered to be significant. Statistical analysis was done using graphpad prism 7 software.
Example 3 : RESULTS AND DISCUSSION
PHYTOCHEMICAL ANALYSIS
The phytochemical analysis of various extracts was performed and presence of flavonoids, phytosterol, carbohydrate, terpenoids, Proteins and amino acids were significant.
Table No. 1: Qualitative Phytochemical analysis
S.No Tests Methanolic extracts Petroleum ether Extracts
1 Alkaloids
+
_
2 Carbohydrates
+
+
3 Glycosides + _
4 Saponins _ _
5 Phytosterols
+ +
6 Tannins and phenolic compound
+ _
7 Proteins & amino acids
+ +
8 Flavonoids
+ +
9 Terpenoids
+ +
10 Mucilage _ +
Note: + indicates presence, - indicates absence

INVITRO ANTIOXIDANT ACTIVITY
Nitric oxide radical scavenging activity

Antioxidant activity was evaluated by using nitric oxide radical scavenging activity method. In this study two extracts are used namely methanol and petroleum ether the % inhibition release of nitric oxide is the parameter considered as their antioxidant activity.
The % inhibition of methanol, petroleum ether and standard ascorbic acid values are mentioned in table 3 and Fig. 7&8. the extracts was able to neutralize nitric oxide in a concentration dependent manner at a concentration range of 200- 1000µg/ml and IC50 (µg/ml) value has been calculated and was found to be 213µg/ml for ascorbic acid, whereas methanol and petroleum ether extracts showed the IC50 - value (µg/ml) of 238.9 and 237 respectively. From the studies the methanol and petroleum ether extracts have significant antioxidant activity in comparison to the standard (ascorbic acid).
Recent studies show that antioxidants are responsible for antianxiety and sedative activity. Hence the extracts are subjected for antioxidant activity and it is found out that all the extracts show antioxidant activity to a certain extent compared to the standard. There forth all the extracts can be further used for in vivo studies.
Table 2: Nitric oxide radical scavenging activity of ascorbic acid, methanol and petroleum ether extracts
S. No.
Concentration(µg/ml) % of scavenging of NO
Ascorbic acid Methanol extract Petroleum ether Extract
1.
200
49.528
48.58
46.16

2.
400
69.974
57.33
65.41

3.
600
77.857
58.14
70.12

4.
800
93.361
62.58
85.46

5.
1000
96.266
72.27
91.11

IC50 (µg/ml)
213
238.9
237

Example 4: RESULTS AND DISCUSSION
INVIVO STUDIES
I ANTIANXIETY ACTIVITY IN MICE USING ELEVATED PLUS MAZE METHOD
The elevated plus maze test is most popular test for evaluation of anxiolytic compounds. The elevated plus maze is highly sensitive to the influence of both anxiolytic and anxiogenic drug acting at the GABAA benzodiazepine complex. The EPM test is used to evaluate the psychomotor performance and emotional aspects of mice. EPM is considered as one of the well-established models for unconditioned anxiety to detect anxiolytic/anxiogenic like activity by investigating aspects of physiological and pharmacological behavior. In EPM, mice will normally prefer to spend much of their allotted time in enclosed arms. This preference appears to reflect an aversion towards open arms that is generated by the fears of the open spaces. In the EPM test increased number of entries and time spent into the open arm are taken as the index/reliable indicators of decreased anxiety or indicating the anxiolytic-like activity of a compound.85-87
The percentage open arms entry with control group was 12.43±1.14 and the percentage time spent in open arms was 7.6±0.67 seconds.
With diazepam 1mg/kg in group II, the percentage open arms entries was 66.81±0.58 and the percentage time spent in open arm was 69.93±0.41 seconds when compared to control groups.
With test group III i.e. Extracts of Naringenin, 50mg/kg, the percentage of open arm entry was 40.55±0.52 and percentage time spent in open arm was 46.73±0.63 second

With test group IV i.e. Extracts of Naringenin, 100mg/kg, the percentage of open arm entry was 50.28±0.9 and percentage time spent in open arm was 56.50±0.39 seconds.
The result extracts of Naringenin seeds in the dose of 100mg/kg was highly significant when compared to control as shown in table 4 and Fig.9&10
Table 3: Effect of Extract of Naringenin in elevated plus maze


S.
No

Treatment group

% open arm entries
% time spent in open arm

1
Control
12.43±1.14
7.6±0.67

2
Diazepam 1mg/kg 66.81±0.58*** 69.93±0.41***

3
Naringenin50mg/kg
41.55±0.52*** 47.73±0.63***

4
Naringenin 100mg/kg 50.28±0.9*** 56.50±0.39***
All values are expressed as Mean±SEM(n=6). One way ANOVA followed by Dunnet's test. *** P?0.001 when compared to control.
In the present study, mice treated with methanol and petroleum ether extracts of Naringenin at the doses of 100mg/kg produced significant (P?0.001) anxiolytic effects. In the EPM test when compared to control as evidenced by increased percentages of both open arm entries and time spent in open arms when compared to control group of animals.
II SPONTANEOUS LOCOMOTOR ACTIVITY
Spontaneous locomotor activity is considered as an index of alertness and can be helpful to confirm the general depressive activity of any drug. The decrease in motor activity gives an indication of the level of depression of CNS.88
Since, an anxiolytic also produces sedation and hypnosis; these activities were evaluated with spontaneous locomotor activity in Actophotometer and open field test.
1) Actophotometer
Actophotometer registers the number of times photo beams of light was broken as the mice moved around inside the cage. Each mice was placed in the centre of the metal cage of actophotometer and its ambulatory activity was measured for 10 minutes.89
Locomotor activity is evaluated by using Actophotometer. The spontaneous locomotor activity made by a mouse was noted in control, standard and test group before and 30 and 60 min after the administration of control, standard and test drugs.
The average number of counts at before and 30 and 60 min after the administration of control group of mice was 196.17±2.6, 190.67±2.38 and 189.83±2.5 respectively.
The average number of counts at before and 30 and 60 min after the administration of standard group of mice was 199.17±3.0, 117.5±2.2 and 80.5±2.6 respectively.
The average number of counts at before and 30 and 60 min after the administration of test group (methanol extract) of mice was 192.16±3.0, 162.17±1.6 and 135.83±2.4 respectively.
The average number of counts at before and 30 and 60 min after the administration of test group (pet. Ether extract) of mice was 181.67±2.49, 153±2.9 and 119.3±1.8 respectively.
In actophotometer test, number of cut off (crossing) decreases in test groups compared to control group .The results were shown in table 5.
Table 4: Locomotor activity of mice in actophotometer


S.No
Treatment group Before drug
Administratio n
after 30 min
after 60 min

1
Control
196.17±2.6
190.67±2.38
189.83±2.5

2
Diazepam 1mg/kg
199.17±3.03
117.5±2.24***
80.5±2.6***

3
Naringenin 50mg/kg
192.16±3.07
162.17±1.58*
135.83±2.4***

4
Naringenin100mg/kg
181.67±2.49
153±2.9**
119.3±1.8***

All values are expressed as Mean±SEM (n=6). One way ANOVA followed by Dunnet's test. *P<0.05, **P<0.01 and *** P?0.001 when compared to control.
In the present study, mice treated with methanol and petroleum ether extracts of Naringenin at the dose of 100mg/kg showed A statistically significant (P<0.001) reduction in spontaneous locomotor activity in astrophotometer after 60 minutes of administration of standard and test compounds were noted in comparison with control group of animals that signifies sedative activity.
CONCLUSION
Preliminary phytochemical investigation was done for the Naringenin It was found to contain flavonoids.
The present study shows that the Naringenin have higher antioxidant activity which was determined using the method (Nitric oxide radical scavenging activity).
The study shows that extracts of Naringenin at the dose of 100mg/kg has significant antianxiety activity.
The study also shows that the antianxiety of various extracts of Naringenin at the dose of 100mg/kg is less efficacious when compared to the standard drug diazepam at the dose of 1mg/kg.
The results from the experiments confirmed that the extract of Naringenin posses a strong anxiolytic potential. However, the further studies in other models and extensive phytochemical analysis are necessary to identify the exact chemical compound and its possible mechanism of action underlying the anxiolytic and effect of Naringenin
Anxiety related disorders are the most common mental illness and a major cause of disability in the world. Mental disorders have been found to be common, with over a one third of people in most countries reporting them with sufficient criteria to be diagnosed at one point in their life.
Anxiety is a frequent problem that affects people of all the ages around the world. It is a prevalent and potentially serious condition that adversely affects the diurnal functioning, health status and life quality of people of all of age. Stress, anxiety and depression could trigger insomnia.
Despite a phenomenal development of modern drug industry, medicinal plants still constitute an important part of pharmacopoeias in both the developed and developing countries. These plants are important elements of traditional medicine and can be developed as potential drug after scientific validation. However, many of these traditionally used plants have not yet been studied scientifically.91
Naringenin the results of Preliminary phytochemical investigation shown the presence of various phytochemical constituents like flavonoids.

While considerable emphasis has been placed herein on the specific features of the preferred embodiment, it will be appreciated that many additional features can be added and that many changes can be made in the preferred embodiment without departing from the principles of the disclosure. These and other changes in the preferred embodiment of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.

, Claims:We Claim,
1. A method for treating anxiety and related disorders in mammals, comprising: administering a therapeutically effective amount of Naringenin or its pharmaceutically acceptable extract, wherein the administration results in:
a) modulation of GABA receptor activity to reduce anxiety symptoms;
b) scavenging of nitric oxide radicals to mitigate oxidative stress; and
c) a dose-dependent sedative effect, as determined by reduced locomotor activity in behavioral models.
2. A pharmaceutical composition for treating anxiety and inducing sedation, comprising:
a) Naringenin extract derived from natural sources in an effective concentration; and
b) a pharmaceutically acceptable carrier or excipient, wherein the composition exhibits significant antioxidant activity as measured by nitric oxide radical scavenging assays with IC50 values ranging between 230-250 µg/mL.
3. The method claimed in claim 1, wherein the method of evaluating the anxiolytic and sedative properties of Naringenin, comprising:
a) administering Naringenin at doses of 50-100 mg/kg body weight to subjects;
b) assessing anxiety reduction using an Elevated Plus Maze (EPM) by measuring increased time spent and entries into open arms; and
c) evaluating sedation through decreased locomotor activity in an Actophotometer test, wherein the results demonstrate a statistically significant improvement in anxiolytic and sedative effects compared to control groups.
4. The method claimed in claim 1, wherein the use of Naringenin in the manufacture of a medicament for the treatment of anxiety-related disorders and oxidative stress-induced conditions, wherein the medicament is formulated to deliver doses effective for modulating oxidative stress and enhancing GABA receptor activity.
5. The method claimed in claim 1, wherein the method of reducing anxiety and oxidative stress simultaneously, comprising: administering Naringenin extract obtained through methanolic or petroleum ether extraction, wherein the extract is effective in both:
a) increasing open-arm entries in an Elevated Plus Maze test; and
b) scavenging nitric oxide radicals with antioxidant efficacy comparable to standard ascorbic acid.

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