INTRODUCTION:
Everyone is impacted by the ubiquitous phenomenon of stress. Excessive stress is harmful to the body and must be managed as such. Stress has an impact on the onset of many diseases, including diabetes mellitus, peptic ulcers, male impotence, immunosuppression, and psychiatric disorders like anxiety and depression.
Biogenic amines like adrenaline and nor-adrenaline have been demonstrated to cause a significant rise in brain levels while under stress.1 The WHO estimates that 450 million people worldwide suffer from mental or behavioral problems like stress, which account for 12.3% of the world's illness burden.2
Stress is the body's general reaction to any demands placed upon it. Numerous ailments, such as hypertension, peptic ulcers, immunosuppression, reproductive dysfunction, and behavioral disorders, are associated with stress.3 The body's natural response to unfavorable circumstances, such as trauma or mental upheavals, is stress. This response is made up of acts that typically endanger or disturb the organisms' homeostasis.4 Chronic stress is linked to altered levels of monoamine and 5-hydroxytryptamine, which are important stress-related chemicals. Hormone fluctuations result in abnormal behavior and disruption of the hypothalamus-pituitary-adrenal (HPA) axis. These disorders can include anxiety, depression, OCD, eating and sleeping disorders, hyperglycemia, and weakened immune response.5 Modern drug therapy for diseases related to stress does not offer a perfect cure. Increasing endurance through available methods, yoga, medication, nutraceutical supplementation, and adaptogen intervention. The last ten years have seen a significant amount of research on the effects of stress on biochemical, neurochemical, and molecular mechanisms.6,7,8,9,10 According to recent development have reported that the episode of depression is enhanced to 5 to 6 folds following of the occurance of stressful “life events”. The stress is are consider among the most prevalent psychiatric or mental disorder or syndrome, aaffects the 10% to 30% of general populations of the industrialized areas.11,12
The term "adaptogen" refers to an agent that increases an organism's ability for adaptation during stress, and the word "antistress" (a pharmaceutical name for the same thing) refers to an agent that both promotes adaptation and neutralizes or prevents the negative consequences of stress.13 The present study aims to investigate antistress potential of leaves Sida veronicafolia (Lam).
Traditional medicine is the body of knowledge and methods used to cure a variety of physical and mental ailments, as well as to maintain health, that are derived from the theories and experiences of many civilizations. In general, the most popular traditional treatment for a variety of illnesses is herbal medicine.14 The majority of the pharmaceutical industry sources its secondary metabolites from medicinal plants in order to create human health care products. In India, there is a growing demand for products created from plants because of their safety and medical benefits.15 Sida veronicafolia (Lam) is a well-known herb used in Ayurvedic and Siddha treatment. The plant is also referred to as Nagabala, Bhumibala, Shaktibala, etc. in Ayurveda.16 A little, thin, straggling plant that roots at nodes and frequently covers with the superficial soil is Sida veronicafolia (Lam.), a member of the Malvaceae family. Measurements: 2–5 cm long, up to 4cm broad, broadly oval, serrated. With unicellular and multicellular trichomes that irritate the skin, the stem and leaves are both highly hairy.17 Although, Antioxidant activity18, Hypoglycemic activity19, Antitumor activity20, Hepatoprotective activity21 properties of Sida veronicafolia (Lam.) have been reported. Therefore, the current study's objective was to find out whether ethanolic extract of Sida veronicafolia (Lam) leaves could reduce stress in rats and mice by protecting them from chemically induced stress, immobilization stress models, and anoxia stress tolerance.
MATERIALS AND METHODS:
Plant material:
Bulk Sida veronicafolia (Lam) leaves were gathered from our college campus and verified by the Y.C. Institute of Science's Department of Botany in Satara. Distilled water was used to wash the leaves of Sida veronicafolia (Lam) in order to get rid of garbage and dirt. It was then coarsely ground after more shade drying.
Preparation of Sida veronicafolia (Lam) leaves extract:
Sida veronicafolia (Lam) leaf powder was extracted using increasing polarity solvents in a series of soxhlet extractions. For this study, the ethanolic extract was chosen. A rotary flash evaporator was used to concentrate the extract, which was then kept at room temperature.
Route and Dose Administration:
An earlier report22 stated that the EELSVL was safe and that there was no animal mortality. Therefore, using the fixed dosage approach of CPCSEA, 2500 miligram per kilogram was determined to be the LD50 cut-off value. Therefore, for the Antistress action, the dosages of 250 miligram per kilogram per oral and 500miligram per kilogram per oral were chosen.
Animals:
For this investigation, healthy albino wister mice weight is 20–30g and albino wister rats weight is 150–225g were employed. The mice and rats were kept under conventional circumstances, with 12hours of light and dark cycles, 25±300C temperature, in polypropylene cages, with a humidity range of 35 to 60%. They have unlimited access to food and drink.
Evaluation of Anti-stress activities:
Anoxia stress tolerance test in mice:
Twenty to thirty grams of albino mice, of either sex, were chosen, and they were split into the 4 groups of 6. Group I was given purified water (1 milliliter per kilogram, p.o.), Group II was given an ethanolic extract of leaves of Sida veronicafolia (Lam) (EELSVL) (250 mg per kilogram, p.o.), Group III was given an ethanolic extract of leaves of Sida veronicafolia (Lam) (500 mg per kilogram, p.o.), and Group IV was given diazepam (2 milligrams per kilogram, i.p.). The animals received treatment for the three weeks that were mentioned above. In order to record the anoxia tolerance time, stress was induced at the end of the first, second, and third week by individually placing each animal in a hermetic vessel with a capacity of one liter on the seventh, fourteenth, and twenty-first day, one hour after the treatment. As soon as the animal began to convulse, the transit took it from the vessel and, if necessary, performed Treatment. The animal's anoxia tolerance was measured by timing the animal's entrance into the hermatic vessel and the onset of the first convulsion. The animal's apparent convulsion was a very sharp end point because if it is not removed from the vessel within a minute, it could die.23
Chemical induced stress in mice:
Male or female albino mice weighing 20–30 g were split into four groups of six. The group I was assigned to as a vehicle control was given either distilled water (0.1 ml/100g) or a vehicle. Different or special doses of EELSVL (250, 500mg/kg p.o.) were administered to the animals in Groups II and III. Group IV animals received dizepam treatment (2mg/kg). Each treatment was given nonstop for a period of fifteen days. Day 15 After an hour of medication administration, each animal given 0.1 ml of 6% glacial acetic acid (v/v) intraperitoneally. For twenty minutes, the number of writhes in each group was recorded.24,25
Immobilisation Stress test in rats:
Five sets of six adult albino rats weighing between 150 and 225 grams each were chosen. The group I was designated negative control (untreated, unstressed). Group II was designated as the positive control group (unstressed, got vehicle), EELSVL was administered to group III 250 miligram per kilogram by per oral, group IV received EELSVL 500 miligram per kilogram by per oral and 2mg/kg Diazepam received group V by intraperitonally. The above mentioned treatment created or was regularly given for ten days, starting one hour before stress exposure. Stress was created by immobilizing animals in a supine position with their heads down, their hands and forelimbs strapped to a hardwood board that was angled 600 degrees, twice a day for ten days. Stress was induced by placing the animals in a supine position, heads down, twice a day for 10 days, with their hands and forelimbs secured to a 60-degree-angled wooden board.26
Statistical analysis:
The data were evaluated using one-way analysis of variance (ANOVA), Tukey's, and mean ± standard error of mean (SEM) presentation. P<0.05 is considered statistically significant. For the statistical study, GraphPad Prism was utilized.
RESULTS AND DISCUSSION:
Anti-stress Activity:
Effect of EELSVL on the anoxia stress tolerance test:
The anoxia stress tolerance test duration was shown to be considerably longer in the groups of animals treated with EELSVL at doses of the 250 miligram per kilogram (P<0.01) and 500 miligram per kilogram (P<0.001), as well as the groups of animals treated with diazepam at 2 mg/kg (P<0.001) i.p. on the 7th, 14th, and 21st day of therapy. There was a dose-dependent relationship between the effects. The outcomes are displayed in Table I.
Effect of EELSVL on the Chemical Induced Stress test:
0.1 ml glacial acetic acid of 6% (v/v) concentration either significantly enhanced quantity of writhing in albino mice. Prior the treatment 250mg/kg EELSVL (P<0.05) was shown to significantly inhibits the numbers of writhes in twenty minutes when the compared to the control group. In 20 minutes, there were significantly inhibits writhes in the EELSVL group treated with a huge dose of 500 miligram per kilogram (P<0.01) than group V diazepam treated with a dose of 2mg/kg (P<0.01). That is to say, EELSVL had a dose- or proportionate-related impact. The outcomes are displayed in Table II.
Effect of EELSVL on the immobilisation stress test :
Effect of EELSVL on the biochemicals parameters:
Changes of physiological (biochemical) parameters such as blood urea nitrogen, glucose, and cholesterol rise in rats under immobilization stress, causing changes in these parameters. Nevertheless, blood urea nitrogen (P<0.001) and glucose and Cholesterol (P<0.05) dramatically decreased when EELSVL at a dose of 250 mg/kg p.o. was administered. On the other hand, there was a significant (P<0.001) decrease in blood urea nitrogen , glucose, and cholesterol levels after treatment with a large dose of EELSVL 500mg/kg p.o. indicating that the impact was dose related. The outcomes are displayed in Table III.
Effect of EELSVL on the organs weight:
Weight increases in the liver and adrenal glands and significant decreases in the spleen were observed in rats under immobilization stress. Conversely, the spleen's weight grew significantly (P<0.001), while the adrenal glands' (P<0.01) and liver's (P<0.001) and weights dramatically reduced at a dose of 250 mg/kg P.O. of EELSVL. However, the weight of the spleen increased considerably (P<0.001) when EELSVL was given at a high dose of 500 mg/kg p.o., while the weight of the adrenal glands (P<0.01) & liver (P<0.001) and decreased, suggesting that the impact was dose related. The outcomes are displayed in Table IV.
Table I : Effect of EELSVL on the anoxia stress tolerance test.
|
Treatment group |
Anoxia stress tolerance time (minutes) |
||
|
1st week (7th day) |
2nd week (14th day) |
3rd week (21st day) |
|
|
Vehicle control |
133.66 ± 3.30 |
134.43±1.56 |
138.74±0.87 |
|
EELSVL (250 mg/kg) p.o. |
143.59±1.684** |
152.17±1.63** |
159.28±1.73** |
|
EELSVL (500 mg/kg) p.o. |
157.52±3.27*** |
162.10±4.34*** |
169.77±3.60*** |
|
Diazepam (2 mg/kg) i.p. |
180.76±2.372*** |
185.25±2.09*** |
188.23±1.651*** |
*P<0.05, **P<0.01, and ***P<0.001 when compared to the control group (one-way ANOVA followed by Tukey test). The data, with n = 6, are shown as mean ± SEM.
Table II : Effect of EELSVL on the Chemical induced stress test.
|
Treatment group |
The No. of writhe (minutes.) |
|
Vehicle control |
62.16±1.35 |
|
EELSVL (250 mg/kg) p.o |
33.01±1.46* |
|
EELSVL (500 mg/kg) p.o. |
27.33±1.52** |
|
Diazepam (2 mg/kg) i.p. |
14.33±1.22** |
*P<0.05, **P<0.01, and ***P<0.001 when compared to the control group (one-way ANOVA followed by Tukey test). The data, with n = 6, are shown as mean ± SEM.
Table III : Effect of EELSVL on the biochemical parameters in immobilisation stress test.
|
Treatment group |
The biochemical analysis (miligrams per deciliter) |
||
|
Glucose parameter |
Cholesterol parameter |
Blood Urea Nitrogen parameter |
|
|
Un-stressed/Untreated (negative control group) |
86.77± 2.721 |
53.28± 1.85 |
25.07± 1.47 |
|
stressed and vehicle (positive control group) |
151.27± 4.599 |
70.75± 1.21 |
43.22± 1.37 |
|
EELSVL (250 mg/kg) p.o. |
145.09± 2.94* |
65.92± 1.82* |
39.58± 1.33*** |
|
EELSVL (500 mg/kg) p.o.. |
138.2± 1.99*** |
59.91± 1.40*** |
36.008± 1.32*** |
|
Diazepam (2 mg/kg) i.p. |
133.1± 1.45*** |
55.61± 1.19*** |
30.66± 1.24*** |
*P<0.05, **P<0.01, and ***P<0.001 when compared to positive group (one-way ANOVA followed by Tukey test). The data, with n = 6, are shown as mean±SEM.
Table IV: Effect of EELSVL on the organ weight in immobilisation stress in rats
|
Treatment groups |
The organ weight (gm /100 gm b.w) |
||
|
Liver |
adrenal gland |
Spleen |
|
|
Un-stressed/ Untreated (negative control group) |
4.40 ± 0.471 |
0.014 ± 0.001 |
0.39 ± 0.014 |
|
stressed, and vehicle (positive Control group) |
6.30 ± 0.334 |
0.031 ± 0.002 |
0.27 ± 0.024 |
|
EELSVL (250mg/kg) p.o. |
5.65 ± 0.270*** |
0.018 ± 0.001** |
0.35 ± 0.017*** |
|
EELSVL (500mg/kg) p.o. |
5.59 ± 0.181*** |
0.013 ± 0.001** |
0.37 ± 0.019*** |
|
diazepam (2mg/kg) i.p. |
3.86 ± 0.305*** |
0.016 ± 0.001*** |
0.38 ± 0.009*** |
*P<0.05, **P<0.01, and ***P<0.001 when compared to the control group (one-way ANOVA followed by Tukey test). The data, with n = 6, are shown as mean ± SEM.
DISCUSSION:
The antistress activity of an ethanolic extract of Sida veronicafolia (Lam) leaves against immobilization stress models, chemically induced stress, and anoxia stress tolerance has been investigated in the current work. Diazepam is a commonly given medicine due to its non-specific antistress effect that involves the mesocortical dopamine system, norepinephrine, and 5HT levels of the entire brain and hypothalamus. It has been proposed that increased GABAergic neurotransmission could be the source of this effect. 27
Reduced or inhibited oxygen in the hermatic vessel in anoxia stress tolerance models causes convulsions in the animals; however, the duration of stress tolerance was raised or prolonged prior to treatment with an ethanolic extract of Sida veronicafolia (Lam) leaves, showing anti-stress activity. One plausible explanation could be that, in times of stress, the ethanolic extract of leaves of Sida veronicafolia (Lam) were able to raise the level of succinate dehydrogenase in the brain. This enzyme is in responsible of energy utilization and conservation in the cellular system of the organism, helping in the adaptive process. Ethanolic leaf extract from Sida veronicafolia (Lam) demonstrated strong antistress action in a chemically induced stress condition, as demonstrated by a considerable decrease in the number of writhing in the animals. Biochemical indicators such blood urea nitrogen, glucose, and cholesterol rise significantly in the positive control group in the immobilization stress model compared to the other groups.
Significant hyperglycemia was seen in the current study, and reports indicate that the adrenal cortex of humans and rats will release cortisol and corticosterone in response to stressful situations. The gluconeogenesis and lipogenesis processes are two ways in which this hypersecretion of cortisol contributes to maintaining of internal balance.28
The ethanolic extract of Sida veronicafolia (Lam) leaves greatly reduces or inhibits the level of hyperglycemia. This may be because it maintains the homeostasis mechanism in the immobilization stress model and inhibits the hyperactivity of the adrenal cortex. Stress increases blood cholesterol by a process that most likely has to do with the hypothalamo-hypophyseal axis (HPA) blocking activity, which causes catecholamines and corticosteroids to be released. This may result in an enhancement in the scientific understanding of blood cholesterol levels. It is known that adrenaline mobilizes fat (lipids) from adipose tissues. Raised blood urea nitrogen and glucose levels are caused by an increase in catecholamine release.29
The ethanolic extract of Sida veronicafolia (Lam) leaves dramatically reduces elevated levels of blood urea nitrogen, cholesterol, and glucose in an immobilization stress paradigm. This effect is dose-dependent, and one potential mechanism may be the normalization of catecholamine levels. In the immobilization stress methodology, weight of the adrenal glands and liver was found to be dramatically raised, whereas weight of the spleen was shown to be greatly decreased. According to theory, when humans are under stress, they go through an adreno-medullary response in which the pituitary gland's β receptors are activated by adrenaline, causing an increase in ACTH secretion. This has the potential to activate the medulla and adrenal cortex, increasing the weight of the adrenal gland. Conversely, cortisol raises the liver's mRNA levels, which causes the liver to enlarge in weight. Furthermore, there are indications that the spleen contracts during stress in order to produce more red blood cells (RBCs), which causes the spleen's weight is reduced.30,31
The animals' organ weight was greatly restored to its original state after the stress-induced modification, and this result was dose-dependently achieved by the ethanolic extract of Sida veronicafolia (Lam) leaves. The control group had a considerable increase in the number of writhes in the chemically induced stress scenario. There have been reports that the stress test with glacial acid made the pain system hyperalgesic.32
All of these findings confirmed that the ethanolic extract of Sida veronicafolia (Lam) leaves had a significant and dose-dependent antistress activity; however, the previous treatment with the extract may have been caused by the suppression of pain and inflammatory processes, as it was found to significantly inhibit the number of writhes.
CONCLUSION:
According to the data collected, treating mice with an ethanolic extract of Sida veronicafolia (Lam) leaves greatly increased or increased their anoxia tolerance period, and this effect was dose-dependent. It also significantly reduced the frequency of writhes in the animals. Additionally, the immobilization stress-related alterations in metabolic parameters and organ weight were reversed in the groups treated with an ethanolic extract of Sida veronicafolia (Lam) leaves. The normalization of catecholamine levels is considered to be the mediating mechanism underlying this action. The result obtained from this study is suggest that the extract is a potential for the Antistress activity.
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Received on 16.07.2024 Revised on 07.09.2024 Accepted on 27.11.2024 Published on 10.12.2024 Available online on December 17, 2024 Asian J. Res. Pharm. Sci. 2024; 14(4):353-358. DOI: 10.52711/2231-5659.2024.00056 ©Asian Pharma Press All Right Reserved
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