ANTIOXIDANT DRUG USED IN THE TREATMENT OF RESERPIN INDUCED PARKINSON DISEASE IN RATS

ANTIOXIDANT DRUG USED IN THE TREATMENT OF RESERPIN INDUCED PARKINSON DISEASE IN RATS

N. Tyagi ^* and R. Shukla

School of Pharmaceutical Science, Shri Venkateshwara University Gajraula, Amroha - 244236, Uttar Pradesh, India.

ABSTRACT: Parkinson's infection is a neurodegenerative issue portrayed by the cardinal manifestations of solidness, resting tremor, gradualness (bradykinesia) and decrease of development (hypokinesia). It is a maturing populace and in spite of the fact that there have been a few significant leaps forward as far as the treatment of this crippling illness, for example, drugs levodopa, dopamine [DA] agonists, anticholinergic and medical procedure. Parkinson's ailment is an interminable, dynamic, neurodegenerative confusion with an expected pervasiveness of 31 to 328 per100, 000 individuals around the world. It is evaluated that more than 1 percent of the populace over age 65 are burdened with Parkinson's infection, occurrence and pervasiveness increment with age. So, the point of the investigation to assess the impact of daidzein on Parkinson sickness instigated by the reserpine model in rodents. Reserpine is the antihypertensive specialist, incites the consumption of focal catecholamines stores. Infusion of reserpine in rodents causes hypokinesia, unbending nature, tremors, and idleness. Cell reinforcements assume a fundamental job in the avoidance or treatment of Parkinson infection, Daidzein is a cancer prevention agent that extinguish the free radicals. Parkinson was assessed by social tests, for example, the rota street test. Estimation of psychological debilitation was finished by different biochemical estimations to be specific Lipid peroxides (in cerebrum), Protein estimation utilizing Folin's reagent and Brain decreased glutathione estimation. Every one of the outcomes was then contrasted with the standard medication carbidopa + Levodopa (30mg/kg).

Daidzein, Reserpine, Carbidopa + Levodopa, Parkinson

INTRODUCTION: Parkinson's Disease is a confusion of the focal sensory system, including fundamentally a degeneration of certain nerve cells in profound pieces of the mind called the basal ganglia, and specifically lost nerve cells (or neurons) in a piece of the brainstem called the substantia nigra.

These cells make the neurochemical ambassador dopamine, which is mostly in charge of beginning a circuit of messages that organize ordinary development ¹. PD is a gradually dynamic neurodegenerative ailment described by Pill moving tremors, Akathisia (failure to sit still), unbending nature, resting tremor, kinesis (akinesia, dyskinesia), "solidifying" and loss of postural reflexes, unstable (stooped) pose.

No arm is swinging in musicality with legs, sialorrhoea, oculogyric crisis (eyes are held fixed for a variable period) nervous wretchedness, involuntary tremors, seborrhea, and masked outward appearance.

The term Parkinsonism applies for an illness state having such normal attributes, while Parkinson's sickness (loss of motion agitans) is confined to the essential or idiopathic parkinsonism ². PD is a gradually dynamic parkinsonian disorder that starts guilefully and, for the most part, influences one side of the body before spreading to include the opposite side. Parkinson's infection is a ceaseless, dynamic, neurodegenerative turmoil with an expected pervasiveness of 31 to 328 for every 100,000 individuals around the world. It is evaluated that more than 1 percent of the populace over age 65 are distressed with Parkinson's ailment; rate, and pervasiveness increment with age ³. Parkinsonism is a clinical disorder including mixes of engine issues to be specific bradykinesia, resting tremor, unbending nature, flexed stance, "solidifying" and loss of postural reflexes.

Parkinson's sickness is a significant reason for parkinsonism. Parkinson's Disease is a gradually dynamic parkinsonian disorder that starts deceptively and, for the most part, influences one side of the body before spreading to include the opposite side ⁴. Oxidative pressure characterized as an irregularity between biochemical procedures prompting generation of receptive oxygen species and the cell cancer prevention agent course causes atomic harm that can prompt a basic disappointment of organic capacities and eventually cell passing ⁵. In Parkinson's illness, oxidative pressure actuated by free radicals harms neuronal film lipids, proteins, and different segments of mind tissue. There are a few potential wellsprings of the expanded free extreme generation in Parkinson ailment, including mitochondrial brokenness, expanded free iron levels, and expanded dopamine digestion ^6-7.

The Imbalance between cholinergic activity and dopaminergic activity in the striatum causes a neurological disorder such as Parkinson's disease. Dopamine is released in many areas in the brain. The mesolimbic pathway is involved in reward and reinforcement; mesocortical pathway helps to regulate emotional response and motivation. In the central nervous system, acetylcholine helps with attention, arousal, reward, and sensation; loss of dopamine tilts the balance towards too much acetylcholine contributes to motor systems ⁸. Backing for oxidative pressure components in dopaminergic degeneration in the substantia nigra in Parkinson's sickness, ⁹ originates from a developing assemblage of proof, showing that this area has a high inclination for oxidative pressure and is additionally insufficient in defensive components ¹⁰. Be that as it may, when it is in abundance in the cytosol, at that point it is effectively utilized by monoamine oxidase (MAO) to create hydrogen peroxide or via autoxidation to frame quinones ¹¹. Cell reinforcement security of the cerebrum is given by Superoxide dismutase, catalase, and glutathione peroxidase. Glutathione peroxidase is the most powerful compounds that secure against oxygen poisonous quality by searching H₂O₂ created by cell digestion, which is recognized solely in glial cells of the midbrain. The recuperation of diminished glutathione is performed by a decrease of oxidized glutathione (GS-SG) by glutathione reductase (GS-SGRd). The level of diminished glutathione in the substantia nigra is diminished ^12-13.

Antioxidants have a potential job in the counteractive action and treatment of Parkinson's illnesses, due to the expanded of the job of free radicals in adding to their pathogenesis. Antioxidants are generally regarded as being safe, three examples of antioxidants - vitamin E, beta-carotene and lipoic acid ¹⁴. The antioxidant helps cells to cope with oxidative stress by effectively quenching free radicals ¹⁵. Daidzein is an isoflavone aglycone and is created in the body from plant isoflavones. Isoflavones are contained in soybean or soy nourishments in two synthetic structures, i.e., aglycones (unconjugated structure) and glucosides (bound to a sugar atom). Maturation or absorption of soybeans or soy items results in the arrival of the sugar particle from the isoflavone glycoside, it might be consumed into the blood or it might be additionally used by intestinal microbes into the metabolites equol and O-desmethyl-angolensin (ODMA). Daidzein is an antioxidant that quenches the free radicals. Daidzein may help prevent breast cancer; Helps prevent osteoporosis, Relief in Menopause, Reduces Cholesterol ¹⁶.

Etiology: The disorder of Parkinsonism (clinical conditions which take after idiopathic Parkinson's illness) may have various causes, for example, arteriosclerosis, diffuse focal sensory system degenerative malady, rehashed head injury, tumor, metabolic imperfections, for example, Wilson's ailment, substantial metal, or carbon monoxide harming. Medication incited Parkinsonism results from dopamine receptor to hinder by medications, for example, phenothiazines, butyrophenones, and metoclopramide. The perception of parkinsonian side effects in heroin addicts who incidentally utilized 1-methyl-4-phenyl-1, 2, 5, 6-tetrahydro-pyridine (MPTP), a pethidine simple, has prompted the advancement of a helpful creature model for the disorder ¹⁷. A pandemic of encephalitis lethargica during the 1920s was in charge of an episode of early beginning parkinsonism with related serious unbending nature and respiratory confusion.

Even though the etiology of Parkinson's ailment is unknown, it has, for quite some time been theorized that neurodegeneration is instigated by hereditary, ecological, or irresistible issues. Age is the absolute most steady hazard factor and it has been evaluated that there is a combined lifetime danger of one out of 40 for building up Parkinson's illness ¹⁸. Loss of pigmented cells in the substantia nigra is the most steady in Parkinson's illness and ordinarily, the amount of nigral cells decreases from 425 000 to 200 000 at 80 yr. In Parkinson's illness, the substantia nigra shows checked consumption of cells (<100,000) with substitution gliosis. Likewise, tyrosine b-hydroxylase, the rate-restricting advance in dopamine combination, additionally lessens. The rest of the cells contain the profoundly trademark eosinophilic Lewy bodies. The example of nigral cell misfortune in Parkinson's infection contrasts from that because of ordinary maturing.

In Parkinson's malady, cell misfortune is dominatingly from the ventrolateral level of the substantia nigra, yet this locale is generally saved in ordinary subjects ¹⁹. On the off chance that maturing is the essential instrument that records for the time course of practical decay, it doesn't enough clarify the immense between individual changeability in rates of clinical movement of the illness ^{20, 21}. In the wake of maturing, a family ancestry is the most grounded indicator of an expanded danger of building up the ailment, in spite of the fact that the job of a typical situation should likewise be considered ²². Moderately high rates of concordance have been recognized among monozygotic twins when one twin had youthful beginning malady ²³.

The vast majority of the accessible proof backings an autosomal overwhelming legacy of the ailment, yet numerous patients don't demonstrate this example of legacy. The disclosure of two particular changes in the a-synuclein quality (SNCA) situated on chromosome 4q seemed, by all accounts, to be a noteworthy leap forward ^{24, 25}. Alpha-synuclein is a very rationed, bottomless 140 amino corrosive protein of obscure capacity that is communicated mostly in presynaptic nerve terminals in the cerebrum ²⁶. It gives off an impression of being a noteworthy part of Lewy bodies, the trademark intracytoplasmic eosinophilic consideration bodies, and its event in Parkinson's infection recommends that this sickness likewise has a place with those conditions, for example, Alzheimer's, owing to harmful protein conglomeration ²⁷.

Changes in a-synuclein were accounted for in various random families with a solid family ancestry of Parkinson's malady; however, a few investigations have neglected to find this hereditary deformity in different families or sporadic cases, recommending that Parkinson's ailment is just once in a while brought about by such imperfections ²⁸. Thus, endeavors to ensnare the detoxifying com-pounds, debrisoquine 4-hydroxylase (CYP2D6) or N-acetyltranferase 2, in the pathogenesis of Parkinson's infection have been uncertain ^{29, 30}. Parkinson's infection was portrayed during the mechanical insurgency proposing that natural poisons may assume a job in its pathogenesis. The disclosure of MPTP-instigated parkinsonism loans further weight to this hypothesis ³¹.

A provincial domain has been related to an expanded danger of building up Parkinson's illness, recommending that specialists, for example, herbicides or pesticides may have an aetiological job, in spite of the fact that this is restricted to around 10% of patients with Parkinson's sickness ³². Cigarette smoking has been shown consistently to reduce the risk of developing Parkinson's disease ^{33, 34}, even though this might be limited to those with a moderately young age at the beginning of the infection ³⁵. This impact has been credited to a restraint of monoamine oxidase type B by the results of tobacco ignition. Proof connecting dietary variables to Parkinson's malady is uncertain, albeit one investigation has demonstrated a lower nutrient E admission in patients with Parkinson's infection contrasted and controls ³⁶. In general, most instances of Parkinson's infection are probably going to result from a mix of hereditary and ecological elements, and these vary between people. Hereditary changes may incline patients to build up Parkinson's illness whenever joined with other quality transformations or environmental components ³⁷.

Levodopa/Carbidopa (Sinemet): Levodopa (additionally called L-dopa) is the most usually recommended and best medication for controlling the side effects of Parkinson's infection, Levodopa is moved to the nerve cells in the mind that produce dopamine. It is then changed over into dopamine for the nerve cells to use as a synapse. Levodopa enters the cerebrum and is changed over to dopamine while carbidopa expands its viability and averts or diminishes a significant number of the symptoms of levodopa, for example, sickness, spewing, and infrequent heartbeat unsettling influences. It is by, and large suggested that patients take Sinemet on an unfilled stomach, in any event, 30 min prior, or one hour after suppers. There are two types of Sinemet, controlled-discharge or quick discharge Sinemet. Controlled-discharge (CR) Sinemet and prompt discharge Sinemet are similarly compelling in treating the indications of Parkinson's infection, however, a few people lean toward the controlled discharge variant. While Sinemet is the best prescription and has the least momentary reactions, it is related to dangers of long haul symptoms, for example, automatic development's dyskinesia.

Levodopa may likewise cause anxiety, disarray, or unusual developments. Changes in the sum or timing of the portion will normally anticipate these reactions, yet most specialists currently prescribe beginning with options to Sinemet, for example, the dopamine agonists, and use Sinemet when the choices neglect to give adequate alleviation. Taken alone, L-dopa causes queasiness. It experiences quick catabolism by fringe decarboxylase, shaping dopamine, which can't cross the blood-cerebrum hindrance.

L-dopa is, this way, constantly given with a fringe decarboxylase inhibitor (PDI), which diminishes queasiness and breaking points fringe digestion of L-dopa, enabling a little rate to cross the blood-cerebrum hindrance in unblemished structure. In the cerebrum, L-dopa is changed over to dopamine by decarboxylase that is put away in the dopaminergic neurons of the substantia nigra ³⁸.

MATERIALS AND METHODS:

Experimental Design:

Oral Administration of Drugs: Medications were suspended to wanted focus in CMC in saline and managed orally. Equal volumes of CMC in saline was given to control gatherings. Every one of the medications was given in volumes of 10 ml/kg.

Dose: Daidzein was regulated at a portion of 50 and 100 mg/kg ³⁹. L-dopa + Carbidopa was controlled at a portion of 30 mg/kg ⁴⁰. The medication treatment was allowed for 5 days and perceptions were made on the fifth day after medication treatment. The perceptions were set aside a few minutes of the pinnacle impact of the medications (8-11 h).

Experimental Protocol: The test convention was separated into the following gatherings. In this trial, the accompanying gatherings of six rodents every controlled medication once day by day for the length of 5 days. Every one of the gatherings would experience every one of the parameters. Toward the finish of every treatment, the rodents were euthanized for the gathering of cerebrum tissue for biochemical estimation.

TABLE 1: EXPERIMENTAL PROTOCOL

Reserpine Model:

Reserpine-Induced Parkinson Disease: Reserpine the antihypertensive operator incites exhaustion of focal catecholamines stores. Infusion of reserpine in rodents causes hypokinesia, unbending nature, tremors, and idleness. The creatures were treated with reserpine (5 mg/kg, i.p., for 5 sequential days). After 24 h of last treatment creatures were tried for acceptance of seriousness of tremors by giving the scores as pursues: No tremors-0, incidental jerks 1, moderate or discontinuous jerks 2, proceeds with tremors-3. The quantity of tremors was meant 5 min.

On the off chance that creatures were not appearing, at that point 0 scores was be given, on the off chance that creatures indicated 1 or 2 tremors, at that point, 1 score was given, creatures demonstrated 3 or 5 tremors in 5 min then 2 scores was given and for at least 6 tremors, score was given 3. Akinesia was controlled by holding the tail of creature and putting the front paws on the stage and let the creature to walk while holding (number of steps taken with forelimbs of creature was meant 3 min) and strong unbending nature was dictated by suspending the creature with forelimbs on centerpiece of flat glass pole (0.5 cm width) at the stature of 25 cm over the tabletop and time to fall on the base surface was estimated. The cut-off was kept for 1 min. The creatures were treated with daidzein (100 mg/kg, p.o., individually), or L-dopa-carbidopa (30 mg/kg, p.o.) 60 min before the organization of reserpine for 5 back to back days ⁴¹.

Locomotor Activity: After the assessment of tremors, akinesia, and strong unbending nature, locomotor movement was assessed by utilizing actophoto-meter. The contraption comprisess of photoelectric cells, which are associated in the circuit with a counter. At the point when the light emission falling on the photocell is cut off by the creature, a check was recorded for 10 min ⁴².

Grip Strength: Thusly inactivity to hold quality was assessed by utilizing rota bar contraption. The pole is 75 cm long and 3 cm in measurement, partitioned into six areas by plastic plates balanced at the 50 cm over the tabletop. Every creature was put on turning pole (20 rpm), and dormancy to tumble down was recorded in various gatherings of creatures.

Biochemical Estimations:

Lipid Peroxides (in Brain): ⁴³

Principle: Lipid peroxidation is a free radical intervened occasion. The essential results of such harm are an unpredictable blend of peroxides which then breakdown to deliver carbonyl exacerbates The malondialdehyde (MDA) is one such carbonyl that structures a trademark chromo-genic adduct with two atoms of thiobarbituric corrosive (TBA). The calorimetric response of TBA with MDA, an optional result of lipid peroxidation has been broadly adjusted as technique for estimating lipid peroxidation.

Reagents:

0.8% Thiobarbituric Corrosive (TBA) Arrangement: 80 g of TBA was broken up in refined water and the volume was made up to 100 ml.

30% Trichloroacetic Corrosive (TCA) Solution: 30 g of TCA was broken up in refined water and the volume was made up to 100 ml.

KCl Arrangement: 2.42 g of KCl will be broken up in refined water and the volume was made up to 100 ml.

Method: One ml of suspension medium was taken from 10% of tissue homogenate. 1 ml of 30% TCA was added to it, trailed by 1 ml of 0.8% TBA reagent. The cylinders were secured with the aluminum foil and kept in a shaking water shower for 30 min at 80-degree centigrade. Following 30 min, tubes were taken out and kept in super cold water for 30 min. These were then centrifuged at 3000 rpm for 15 min.

The absorbance of the supernatant was perused at 535nm at room temperature against the fitting clear. Clear comprises 1ml refined water, 1ml of 30% TCA, and 1ml of 0.8% TBA.

Calculation: The substance of MDA communicated as n moles framed per mg of protein in the tissue was determined to utilize the recipe:

Focus = A × V / E × P

Where An is absorbance.

V is the vol. of arrangement.

E is elimination coefficient (1.56×10^-6 m^-1cm^-1).

P is the protein substance of tissue determined as mg protein/gm.

Protein Estimation Using Folin’s Reagent: ⁴⁴

Principal: Protein responds with the folin's ciocalteau phenol reagent to give shaded complex. The shading so shaped is because of the response of antacid copper with the protein as in the biurate test and the decrease of phosphomolybdate by tyrosine and tryptophan present in the protein.

Reagents Required:

Basic Sodium Carbonate Arrangement: 100 ml of 0.1 N NaOH arrangement was set up by dissolving 400 mg of NaOH in refined water and the volume was made up to 100 ml. At that point, 2 g of Na₂CO₃ was broken down in 100 ml of 0.1 ml NaOH.

Copper Sulfate Sodium Tartrate Arrangement: 500 mg of CuSO₄ was broken up in 100 ml of refined water and blended it with 1000mg of Na-K tartarate which is disintegrated in 100 ml of refined water.

Soluble arrangement arranged upon the arrival of utilization by blending 50 ml of the reagent 1 and 1 ml of reagent 2.

Folin's Ciocalteau Phenol Reagent: The business reagent will be weakened with 2 volumes of refined water upon the arrival of utilization.

Standard Protein: Bovine serum egg whites arrangement (2 mg/ml): - 10 ml of cow-like serum egg whites will be broken down in 5 ml refined water to get an answer of 2 mg/ml of protein.

Method: 5 ml of antacid arrangement was evaluated to 1 ml of suspension from the supernatant after centrifugation of the 10% tissue homogenate at 3000 rpm and permit representing 10 min. 0.5 ml weakened folin's reagent was included and the cylinder was shaken to blend the arrangement, following 30 min, the eradication against proper clear at 750 rpm will be recorded.

Preparation of Calibration Standard Curve of Protein: 5 ml of ox-like egg whites arrangement (2 mg/ml) was readied and various volumes were taken in 6 tubes. To all cylinders, refined water was added to make up the volume in each cylinder to 1 ml. The protein fixation in the over 6 cylinders was assessed similarly with respect to the example. A diagram was plotted between the grouping of protein and optical thickness.

Brain Reduced Glutathione Estimation: ⁴⁵ Glutathione in the tissue was evaluated by the strategy for Sedlac and Lindsay (1968) utilizing Ellman reagent.

Reagents:

EDTA (0.2 M): 22.3 gm of EDTA was broken up in 300 ml of warm twofold refined water.

EDTA (0.02 M): 20 ml of the above arrangement was weakened to 200 ml with twofold refined water.

Tris Cushion: 0.4 M (PH 8.9): 24.2 gms of tris support was broken down in 100 ml of twofold refined water. 50 ml of 0.2 M EDTA was added to it and the volume of the arrangement was made up to 500 ml with twofold refined water. The PH of the arrangement was acclimated to 8.9 with (6N HCl).

DTNB (0.01 M): 99 mg of DTNB was broken down in 25 ml of supreme methanol.

Trichloroacetic Corrosive (TCA Half): 50 gm of TCA was broken down in 100 ml of two-fold refined water.

Method: Rodents were yielded by moment beheading. The minds were immediately evacuated and washed with super cold saline. 2 ml of 10% homogenate, which was set up in the KCl arrangement, was taken and include 2.5 ml of 0.02 M EDTA. Shake it vivaciously. Take out 2 ml of the above blend and include 4 ml of virus refined water and 1 ml of half TCA and shake it for 10 min, after the fact the substance was moved to the centrifuged tube and centrifuged at 300 rpm for 15 min.

Following centrifugation, 2 ml of the supernatant will be blended with 4 ml of 0.4 M tris support (PH 8.9). The entire arrangement was blended well and 0.1 ml of 0.01 M DTNB was included, the absorbance was perused within 5 min of expansion of DTNB at 412 nm against reagent clear with no homogenate. For clear readings, rather than 2 ml of homogenate 2 ml of refined water was included.

Calculation: All out GSH (tissue) was determined to utilize the recipe portrayed by Ellman (1959). In this manner, the substance 'C0' of GSH is given by

Co = A × D / E

Where An is the absorbance at 412nm

D is a weakening element

E is the molar elimination coefficient (C = 13000 M^-1cm^-1), Co is the centralization of glutathione.

Statistical Analysis: Every one of the outcomes will be communicated as mean ± standard deviation (SD) trailed by investigation of fluctuation (ANOVA) alongside Turkey's numerous exami-nation test. The p<0.05 will be viewed as factually noteworthy.

Procedure of Brain Extract: The rodents have kicked the bucket with the system of survical separation and after that upper side of the neck was shaved and the skin was cleaned with ethanol. Every single careful instrument was disinfected before the medical procedure. The upper neck of the rodent is dismemberment is made to uncover the cerebrum inside the bone skull of the mind. Put the cerebrum from bone skull with the assistance of for shape and put into the formalin arrangement.

FIG. 1: SUBSTANTIA NIGRA PART IN BRAIN

RESULTS AND DISCUSSION: Parkinson's illness is a neurodegenerative issue portrayed by the cardinal side effects of firmness, resting tremor, gradualness (bradykinesia) and decrease of development (hypokinesia). It is the confusion of the focal sensory system, including fundamentally a degeneration of certain nerve cells in profound pieces of the cerebrum called the basal ganglia, and specifically lost nerve cells (or neurons) in a piece of the brainstem called the substantia nigra. These cells make the neurochemical messenger dopamine, which is partly responsible for starting a circuit of messages that coordinate normal movement.

TABLE 2: SUMMARY OF EFFECT OF DAIDZEIN ON AKINESIA AND TREMOR

TABLE 3: SUMMARY OF EFFECT OF DRUG ON ACTOPHOTOMETER AND ROTAROD

TABLE 4: SUMMARY OF LIPID PEROXIDE METHOD

Normally used antiparkinson drugs for decreasing neurodegeneration activity such as Carbidopa + Levodopa. Levodopa (additionally called L-dopa) is the most generally endorsed and best medication for controlling the indications of Parkinson's malady, Levodopa is moved to the nerve cells in the cerebrum that produce dopamine. It is then converted into dopamine for the nerve cells to use as a neurotransmitter.

Levodopa enters the brain and is converted to dopamine while carbidopa increases its effectiveness and prevents or lessens many of the side effects of levodopa, such as nausea, vomiting, and occasional heart rhythm disturbances. Besides, it is hard to enroll huge no of people for such sort of testing. In this manner, this examination led to assess and approve the impact of Daidzein in Parkinson and too wide information of instrument engaged with Parkinson.

In this examination, the Reserpine model was utilized to instigate Parkinson in wistar rodents. It is the most broadly utilized creature model of Parkinson. Reserpine the antihypertensive operator prompts exhaustion of focal catecholamines stores. Injection of reserpine in rats causes hypokinesia, rigidity, tremors, and immobility. Daidzein is an antioxidant that used to treat Parkinson's, It helps cells to cope with oxidative stress by effectively quenching free radicals.

TABLE 5: PROTEIN ESTIMATION METHOD

TABLE 6: BRAIN REDUCED GLUTATHIONE ESTIMATION

FIG. 2: GRAPH OF LIPID PEROXIDE METHOD. All values were expressed as mean ± S.E.M. (n=5), =P<0.0402 when compared with control group =<0.0003 when compared to Daidzein = P<0.0402 when compared to Carbidopa+ levodopa and P<0.0402 = significant when compared to control group (ANOVA followed by Dunnett’s test).

FIG. 3: GRAPH OF PROTEIN ESTIMATION METHOD. All values were expressed as mean ± S.E.M. (n=5), = P<0.0021 when compared with control group =<0.0021 when compared to Daidzein =P<0.0402 when compared to Carbidopa + levodopa and P<0.0021 = significant when compared to control group (ANOVA followed by Dunnett’s test).

FIG. 4: GRAPH OF BRAIN REDUCED GLUTATHIONE ESTIMATION. All values were expressed as mean ± S.E.M. (n=5), = P<0.4495, when compared with control group, =<0.4495 when compared to Daidzein = P<0.4495, when compared to Carbidopa + levodopa and P<0.4495 = significant when compared to control group (ANOVA followed by Dunnett’s test).

Treatment with Daidzein (50mg/kg) & (100mg/kg) and Combination with Carbidopa + Levodopa (50 mg/kg Daidzein + 30 mg/kg Carbidopa + Levodopa + 5 mg/kg Reserpine) 100 mg/kg Daidzein + 30 mg/kg Carbidopa + Levodopa + 5 mg/kg Reserpine) significantly daidzein and levodopa + c alterations in brain disorder in response to noxious as well as non noxious stimuli suggested this drug can be employed in treatment of Parkinson disease. Ameliorating the excruciating signs and symptoms in epilepsy. However, further research is still required to establish their effectiveness in the treatment of Parkinson.

FIG. 7, 8: DAIDZEIN (50 mg/kg) ALONE TREATED GROUP SHOWED NORMAL NUMBER OF FIBERS AND DECREASING NEURONAL DEGENERATION. DAIDZEIN (100 mg/kg) SHOWED DECREASING NEURONAL DEGENERATION AND NO SWELLING WAS SEEN. THIS WAS MORE RECORDED WITH THE LOWER DOSE OF DAIDZEIN (50 mg/kg)

FIG. 9, 10: (DAIDZEIN 50 mg/kg + 30 mg/kg CARBIDOPA + LEVODOPA + 5mg/kg RESERPINE) AND (DAIDZEIN 100 mg/kg + CARBIDOPA + LEVODOPA + 5mg/kg RESERPINE) SHOWED BETTER RESULT FOR DECREASING NEURONAL DEGENERATION

CONCLUSION: Parkinson Disease is a disorder of the central nervous system, involving primarily a degeneration of certain nerve cells in deep parts of the brain called the basal ganglia, and in particular a loss of nerve cells (or neurons) in a part of the brainstem called the substantia nigra. Parkinson disease is a chronic, progressive, neurodegenerative disorder with an estimated prevalene of 31 to 328 per 100,00 people worldwide. Though there are multiple pharmacological treatment option for parkinson but it is complicated to treat primarily because of involvement of numerous mediators in its Pathophysiology and its resistance to medications. The drugs employed clinically in treatment and management of Parkinson was associated with multiple other adverse effect which additionally make its treatment more difficult thus this research was aimed at examining Daidzein in Parkinson and open new vistas in treatment and management of this disease. Daidzein was found to have positive effect in parkinson induced by reserpine model test of wistar rats. The drug can therefore offer an alternative approach in parkinson state. Conclusively additional investigation is required on other animal models to obtain a dependable oversight of the outcome of Daidzein on Parkinson in actual clinical scenario. In rotarod activity normal control group was found significant effect when compared all groups except reserpine group.  It Show good muscles strength. Daidzein (50mg/kg), Daidzein (100mg/kg). Carbidopa + Levodopa (30mg/kg), DMSO and combination of Carbidopa + Levodopa and Daidzein significantly (P<0.1) increased behavioural attention on fifth day. In GSH activity normal control group (Carbidopa + Levodopa 30mg/kg + 50mg/kg Daidzein + 5mg/kg Reserpine) & (100mg/kg Daidzein + 30mg/kg Carbidopa + Levodopa + 5mg/kg reserpine) showed significantly effect on behavioural as well as biochemical estimation parameters. In histopathological study substantia nigra showed neuronal degeneration (Carbidopa + Levodopa 30mg/kg + 50mg/kg Daidzein + 5mg/kg Reserpine) & (100mg/kg Daidzein + 30mg/kg Carbidopa + Levodopa + 5mg/kg reserpine) showed better result no swalling was seen it was show significant effect.

Effect of Carbidopa + Levodopa and Daidzein on Histopathological Evaluation of Neuropathy: These results depict that the normal functionally of Substantia nigra was maintained in normal control rats resulted in neuron degeneration, whereas rats treated with Daidzein shows milder neuronal degeneration. Pharmacological treatment with Daidzein (50 mg/kg & 100 mg/kg), Carbidopa + Levodopa (30 mg/kg) and the Reserpine induce model pathological changes in brain (Substantia nigra) of rats.

Moreover treatment with combination of Carbidopa + Levodopa & Daidzein. (Carbidopa + Levodopa 30 mg/kg + Daidzein 50 mg/kg, Carbidopa + Levodopa 30 mg/kg + Daidzein 100 mg/kg), (Carbidopa + Levodopa 30 mg/kg + 50 mg/kg Daidzein + 5 mg/kg Reserpine) and (30 mg/kg + Daidzein 100 mg/kg + 5 mg/kg Reserpine) markedly protected changes in brain tissue.

ACKNOWLEDGEMENT: It is a great pleasure for me to acknowledge all those who have contributed toward the conception, origin and nurturing of this project. With a deep sense of gratitude and respect, I thank my esteemed research guide Dr. Rahul Shukla Director in School of Pharmaceutical Science (Shri Venkteshwar University) for her in estimable guidance, valuable suggestions and constant encouragement during the course of this study. At this moment, I thank with deep gratitude to my classmate and friends for their moral support, constant encouragement and patience absolutely needed to complete my entire study. It was the blessing of them that gave me the courage to face the challenges and made my path easier.

I am indebted infinitely to care, support and trust is shown by my parents without whom it would not be possible to complete this research work.

CONFLICTS OF INTEREST: Nil

REFERENCES:

1. Lawrence IG: Parkinson disease hand book, The American Parkinson Disease Association 2010.
2. Sharma HL and Sharma KK: Drug therapy for neurodegenerative disorders. Principles of pharmacology, Paras publishing, 544- 547.T.,
3. Pradeep P and Venkatraman S: International journal of pharmacology screening method 2012; 2: 24-37
4. Stanley F: Description of Parkinson’s disease as a Clinical Syndrome. Ann N Y Acad Sci 2003; 2: 1-14.
5. Sayre M, Smith A and Perry G: Chemistry and Biochemistry of oxidative Stress in Neurodegenerative Disease, Current Med Chem 2001; 8: 721-38.
6. Moore J, West B, Dawson L and Dawson M: Molecular pathophysiology of Parkinson’s disease, Annu Rev Neurosci 2005; 28: 57-8.
7. Gołembiowska K, Dziubina A, Kowalska M and Kamiñska K: Paradoxical effects of adenosine receptorligands on hydroxyl radical generation by L-DOPA in the rat striatum, Pharmacol 2008; 319-30.
8. Matthew and Fox, MD., Neuroscience Dale Purves, 4^th edition (2007).
9. Jenner P, Dexter T, Sian J, Schapira H, Marsden CD: Oxidative stress as acause of nigral cell death in Parkinson’s disease and incidental Lewy body disease. Ann Neurol 1992; 32: 82-87.
10. Dickson W: Linking selective vulnerability to cell death mechanisms in Parkinson’s disease. Am J Pathol 2007; 16-9.
11. Asanuma M, Miyazaki I and Ogawa N: Dopamine- or L-DOPA-induced neurotoxicity, the role of dopamine quinone formation and tyrosinase in a model of Parkinson’s disease. Neurotox 2003; 165-76.
12. Perry L, Godin V and Hansen S: Parkinson's disease, A disorder due to nigral glutathione deficiency. Neurosci Lett 1982; 305-10.
13. Sofic E, Lange W, Jellinger K and Riederer P: Reduced and oxidized glutathione in the substantia nigra of patients with Parkinson's disease. Neurosci Lett 1992; 128-30.
14. Rachel, Lewis and Brian, Lockwood. Current Topics in Nutraceutical Research 2007; 5: 7-18.
15. Sambasiva, Rao M, Lalwani, ND, Watanabe TK and Reddy JK: Cancer Research 1994; 1072-76.
16. Barlow J, RN and NP: Breast Cancer and the Environment Research Centers Community Outreach and Translation Cores, (2002).
17. Langston JW, Ballard P, Tetrud JW and Irwin I: Chronic parkinsonism in humans due to a product of meperidine analogue synthesis. Science 1983; 219: 979-88.
18. Quinn N: Parkinsonism Drecognition and differential diagnosis. Br Med J 1995; 310: 145-50.
19. Fearnley JM and Lees AJ: Ageing and Parkinson's disease: substantia nigra regional selectivity. Brain 1991; 114: 2283-301
20. Hoehn MM and Yahr MD: Parkinsonism: onset, progression and mortality. Neurology 1967; 17: 427-42.
21. Zetusky WJ, Jankovic J and Pirozzolo FJ: The heterogeneity of Parkinson's disease: clinical and prognostic implications. Neurology 1985; 35: 522-6.
22. Semchuk KM, Love EJ and Lee RG: Parkinson's disease: a test of the multifactorial etiologic hypothesis. Neurology 1993; 43: 1173-80.
23. Tanner CM, Ottman R and Goldman SM: Parkinson's disease in twins; an etiological study. JAMA 1999; 281: 341-6.
24. KruÈger R, Kuhn W and MuÈller T: Ala30Pro mutation in the gene encoding a-synuclein in Parkinson's disease. Nat Genet 1998; 18: 106-8.
25. Polymeropoulos MH, Lavedan C and Leroy E: Mutation in the a-synuclein gene identi®ed in families with Parkinson's disease. Science 1997; 276: 2045-7.
26. Burke RE: a-Synuclein and parkin: coming together of pieces in puzzle of Parkinson's disease. Lancet 2001; 358: 1567-8.
27. KruÈger R, Kuhn W and MuÈller T: Ala30Pro mutation in the gene encoding a-synuclein in Parkinson's disease. Nat Genet 1998; 18: 106-8.
28. Riedl AG, Watts PM, Jenner P and Marsden CD: P450 enzymes and Parkinson's disease: the story so far. Mov Disord 1998; 13: 212-20.
29. Langston JW, Ballard P, Tetrud JW and Irwin I: Chronic parkinsonism in humans due to a product of meperidine analogue synthesis. Science 1983; 219: 979-80.
30. Semchuk KM, Love EJ and Lee RG: Parkinson's disease and exposure to agricultural work and pesticide chemicals. Neurology 1992; 42: 1328-35.
31. Hellenbrand W, Seidler A and Robra BP: Smoking and Parkinson's disease: a case control study in Germany. Int J Epidemiol 1997; 26: 328-39.
32. Morens DM, Grandinetti A, Reed D, White LR and Ross GW: Cigarette smoking and protection from Parkinson's disease: false association or etiological clue? Neurology 1995; 45: 1041-51.
33. Tzourip C, Rocca WA and Bretteler MMB: Smoking and Parkinson's disease: an age-dependent risk effect? Neurology 1997; 49: 1267-72.
34. Derijk MC, Breteler MMB and den Breeijen JH: Dietary antioxidants and Parkinson's disease: the Rotterdam study. Arch Neurol 1997; 54: 762-5.
35. Marsden CD and Olanow CW: The causes of Parkinson's disease are being unravelled and rational neuroprotective therapy is close to reality. Ann Neurol 1998; 44: S189-96.
36. Fitzgerald MJT: Basal ganglia. In: Fitzgerald MJT, ed. Neuroanatomy Basic and Clinical, 3^rd London: WB Saunders Company Ltd, 1996: 247-55.
37. Lang AE and Lozano AM: Parkinson's disease: second of two parts. N Engl J Med 1998; 339: 1130-43.
38. Katzung BG: Pharmacologic management of Parkinsonism and other movement disorders. Basic and Clinical pharmacology 10^th edition, Mc Graw Hill Lange, 442-54.
39. Bayer T, Colnot T and Dekant W: Toxi Sci 2001; 205-11.
40. Colpaert FC: Pharmacological characteristics of tremor, rigidity and hypokinesia induced by reserpine in rat. Neuropharmacology 1987; 1431-40.
41. Kulkarni SK: 3^rd, New Delhi, Vallabh Prakashan, Experimental Pharmacology; 2005; 117-18.
42. Vogel HG: 2^nd edition, New York, Springer Publication, Drug Discovery and Evaluation. Pharmacological Assays, 2002; 57.
43. Ohkawah, Ohishin and Yagik: Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 351-58.
44. Lowry OH, Rosenbrough NJ, Farr A and Randall RJ: Biol Chem 1951; 265-75.
45. Sedlak J and Lindsay R: Hx Analyt Biochem 1968; 25.
    

    ---

    How to cite this article:

    Tyagi N and Shukla R: Antioxidant drug used in the treatment of reserpin induced parkinson disease in rats. Int J Pharm Sci & Res 2020; 11(7): 3517-28. doi: 10.13040/IJPSR.0975-8232.11(7).3517-28.

    ---

    All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

    ---