ICMR STS 2018 RESEARCH PROPOSAL
1. Reference ID: 2018-00354
2.Title : Modulation of dopaminergic neurotransmission and motor function by aqueous extract of Curcumin longa in haloperidol induced catatonia in mice.
The Parkinson’s disease (PD) is one of the common neurodegenerative disorders which can also be induced by administration of antipsychotic drugs while treating positive symptoms of schizophrenia.1 The motor disorders like dyskinesia,rigidity and tremors can also result from chronic neuroleptic treatment.2 The neuroleptic induced neurological disorders may not be improved by the supplement of dopamine due to chronic blockade of dopamine D2 receptors by the neuroleptic drugs like phenothiazine compounds. The hypothesis of dopamine receptor supersensitivity proposes that antipsychotic drug treatment causes hypersensitization of dopamine D2 receptors.3 There is no single laboratory model to evaluate parkinsonism and in which a proper evaluation of antiparkinsonian activity can be carried out. However, there is a positive correlation between catatonia in the laboratory animals and the extrapyramidal symptoms produced by neuroleptics in humans.4So, Morpurgo described a direct method to screen the drugs affecting dopamine receptors. He induced catatonia which is a state of neurogenic motor immobility and behavioural abnormality manifestations with stupor.5 The occurrence and irreversibility of this neurological disorder with motor immobility in drug induced parkinsonism has been considered as a major clinical issue in the treatment of schizophrenic patients.6The search for new drugs is needed to avoid such side effects caused while treating schizophrenia.
Bioactive derivatives of plants such as flavonoids, stilbenoids and alkaloids possess potent anti-oxidative and anti-inflammatory properties that improve the mitochondrial function and serve as cognitive enhancers.7 The aqueous extract of Curcumin longa being a herbal product has flavonoid as a bioactive derivative and it also has a tendency of passing through blood brain barrier (BBB), which may prove effective in treating drug induced PD and related neuropsychiatry disorders.
The objective of this study will be-
i) to induce catatonia in mice with haloperidol.
ii) to evaluate the effects of the aqueous extract of Curcumin longa on dopaminergic neurotransmission and motor function in haloperidol induced motor dysfunction.
Type of the study: Experimental animal study
Sample size : 30 mice, 5 groups with 6 mice in each group.
Site of study : Depart of pharmacology, Pondicherry institute of
Study period : June to July, 2018(2 months)
The IAEC approval will be obtained to conduct the study. It is an animal study using Swiss albino male mice. The mice from Central Animal House, PIMS will be used for this study.The aqueous extract of Curcumin longa will be procured from a competent herbal drug manufacturer with complete phytochemical constituents furnished by the manufacturer. Swiss albino male mice weighing 25 to 30 gm and of 60 to 90 days old will be used in this study. This will be maintained in plastic cages under contolled lighting conditions (12h light/12h dark regime), relative humidity(50±5%) and temperature (37±2ºC). They will be fed with pellets and ad libitum access to water. Actophotometer and other instruments necessary in Morpurgo method can be used in the Dept of Pharmacology for this study.
Haloperidol will be administered in the dosage of 1mg/kg of body weight/day/animal4 intraperitoneally for 15 days. The assessment will be done as per Morpurgo scoring method weekly after 24 hours of the last dosage (on 8th and 15th day). The 1st group of 6 mice will be treated with vehicle and rest of the groups will be treated with haloperidol. The 3rd, 4th and 5th groups of mice will be pretreated with the aqueous extract of Curcumin longa 50mg, 100mg and 200mg/kg of body weight respectively as per the protocol listed below:
Control group with 6 mice will be treated only with vehicle (sterile water) for 15 days.
Control group with 6mice will be treated with haloperidol for 15 days.
Experimental group with 6 mice will be pretreated with the aqueous extract of Curcumin longa 50mg/kg of body weight /day/animal intraperitoneally for 15days. Haloperidol will be administered 50 mins after Curcumin longa administration for 15 days.
Experimental group with 6 mice will be pretreated with aqueous extract of Curcumin longa 100 mg/kg of body weight/day/animal intraperitoneally for 15 days. Haloperidol will be administered 50mins after Curcumin longa administration for 15 days.
Experimental group with 6 mice will be pretreated with aqueous extract of Curcumin longa 200mg/kg of body weight/day/animal intraperitoneally for 15 days. Haloperidol will be administered 50 mins after Curcumin longa administration for 15 days.
The locomotor activity and Morpurgo catatonia scoring of control and experimental animals will be measured by using actophotometer and Morpurgo methods respectively on 8th and 15th day before drug administration. The data observed as per the table given above and this will be tabulated for statistical analysis.
Methods of statistical analysis & test applied:
All the results will be expressed as mean ± SEM. Data will be analyzed by one way analysis of variance (ANOVA) followed by Turkey’s multiple comparison test using SPSS 11 software. p<0.05 will be considered as statistically significant. 6.Implications: The Curcumin longa can be used as adjuvant therapy in the treatment of neuropsychiatry disorders such as schizophrenia. This method is emphasised in this study to evaluate antiparkinsonian activity in drug induced parkinsonism by using aqueous extract of Curcumin longa in haloperidol induced catatonia. This can open avenue for adjuvant therapy which can be used with neuroleptic drugs that will avoid untoward effects while treating the schizophrenic patients. Thus the aqueous extract of Curcumin longa can reduce the socioeconomic burdens and side effects caused by existing neuroleptic drugs. This can reduce mortality and morbidity to a greater extent. STAGE BEHAVIOUR SCORE STAGE-0 Mouse moves normally, when placed on the table. 0 STAGE-1 Mouse moves only, when touched or pushed. 0.5 STAGE-2 No movements can be seen in mouse ,even upon eliciting. 0.5 STAGE-3 Mouse placed on the table with one of the front paws raised on a 3cm block, fails to correct the posture in 10 seconds. 0.5 for one paw, therefore 0.5×2=1. STAGE-4 Mouse placed on the table with one of the front paws raised on a 9cm block fails to correct the posture. 1 for one paw = 1×2=2 Table 1: The severity of catatonia induced by haloperidol in mice is scored as follows: By morpurgo modified by Jesudoss et all: The severity of the catatonia will be scored in the mice and it will be checked if the aqueous extract of curcumin longa can reduce the severity and helps in reversing it back. 7.References: 1.Caroff SN, Hurford I, Lybrand J, Campbell EC. Movement Disorders Induced by Antipsychotic Drugs: Implications of the CATIE Schizophrenia Trial. Neurologic clinics. 2011;29(1):127-viii. doi:10.1016/j.ncl.2010.10.002. 2.Mathews M, Gratz S, Adetunji B, George V, Mathews M, Basil B. Antipsychotic-Induced Movement Disorders: Evaluation and Treatment. Psychiatry (Edgmont). 2005;2(3):36-41. . 3.Dhingra, Dinesh,Gahalain, Nidhi. Amelioration of Haloperidol-Induced Orofacial Dyskinesia And Catalepsy By Ellagic Acid In Rats. International Journal of Research in Ayurveda & Pharmacy. Mar-Apr 2016; 7(Suppl 2): 222-227 4. KULKARNI, S.K. & ARZI, A & KAUL, P.N.. (1980). Modification of drug-induced catatonia and tremors by quipazine in rats and mice. Japanese Journal of Pharmacology - JPN J PHARMACOL. 30. 129-135. 10.1254/jjp.30.129. 5.Jesudoss Prabhakar et al. Methods used in screening antiparkinson activity: Errors in published literature. International Journal of Pharmacological Research. 2017; 7(05): 98. 6.Bishnoi et al. (2008). Protective effect of Curcumin, the active principle of turmeric (Curcuma longa) in haloperidol- induced orofacial dyskinesia and associated behavioural, biochemical and neurochemical changes in rat brain. Pharmacology, biochemistry, and behavior. 88. 511-22. 0.1016/j.pbb.2007.10.009. 7.Essa MM, Vijayan RK, Castellano-Gonzalez G, Memon MA, Braidy N, Guillemin GJ.Neuroprotective effect of natural products against Alzheimer's disease. Neurochem Res 2012;37:1829-42.