Kiran Kumar Akula

Potentials of Curcumin as an Antidepressant

Major depression, a debilitating psychiatric disorder, is predicted to be the second most prevalent human illness by the year 2020. Various antidepressants, ranging from monoamine oxidase inhibitors to recently developed dual reuptake inhibitors, are prescribed for alleviating the symptoms of depression. Despite the availability of these blockbuster molecules, approximately 30% of depressed patients do not respond to the existing drug therapies and the remaining 70% fails to achieve complete remission. Moreover, antidepressants are associated with a plethora of side effects and drug-drug/drug-food interactions. In this context, novel approaches are being tried to find more efficacious and safer drugs for the treatment of major depression. Curcumin is one such molecule that has shown promising efficacy in various animal models of major depression. Although the mechanism of the antidepressant effect of curcumin is not fully understood, it is hypothesized to act through inhibiting the monoamine oxidase enzyme and modulating the release of serotonin and dopamine. Moreover, evidences have shown that curcumin enhances neurogenesis, notably in the frontal cortex and hippocampal regions of the brain. The use of curcumin in clinics for the treatment of major depression is limited due to its poor gastrointestinal absorption. The present review attempts to discuss the pharmacological profile along with molecular mechanisms of the antidepressant effect of curcumin in animal models of depression. A need for clinical trials in order to explore the antidepressant efficacy and safety profile of curcumin is emphasized.

Rofecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor increases pentylenetetrazol seizure threshold in mice: possible involvement of adenosinergic mechanism.

Multiple lines of investigations have explored the role of cyclooxygenases (COX) in epilepsy and related neuropsychiatric disorders. Cyclooxygenase particularly, COX-2 expression was found to increase in brain during seizure paradigms. The present study was carried out to investigate the effect of rofecoxib, a selective COX-2 inhibitor against pentylenetetrazol (PTZ i.v.) seizure threshold in mice. The study was further extended to elucidate the possible involvement of adenosinergic mechanism in mediating its anticonvulsant action. Minimal dose of PTZ (i.v., mg/kg) needed to induce different phases (myoclonic jerks, generalized clonus and tonic extension) of PTZ convulsions were noted as an index of seizure threshold. Acute administration of rofecoxib (4mg/kg, i.p.) before PTZ infusion produced an elevation of seizure threshold for all the phases of convulsions. A lower dose of rofecoxib (2mg/kg, i.p.) showed an increase in PTZ seizure threshold for the onset of myoclonic jerks and tonic extension phases but not for generalized clonus. A still lower dose of rofecoxib (1mg/kg, i.p.) failed to increase the threshold in any of the convulsive phases induced by PTZ i.v. infusion. Pretreatment with sub-effective dose of rofecoxib (1mg/kg, i.p.) enhanced the action of sub-protective doses of either adenosine (25mg/kg, i.p.) or 2-chloroadenosine (1 or 2mg/kg, i.p.) in increasing the seizure threshold. On the contrary, treatment with caffeine (100 or 200mg/kg, i.p.) or theophylline (50 or 100mg/kg, i.p.), both non-selective A(1)/A(2) adenosine receptor antagonists reversed the anticonvulsant effect of rofecoxib (4mg/kg, i.p.). Further, dipyridamole (5mg/kg, i.p.), an adenosine uptake inhibitor displayed an anticonvulsant effect with rofecoxib (1mg/kg, i.p.). The study for the first time demonstrated the possible involvement of adenosinergic system in the anticonvulsant effects of rofecoxib against PTZ i.v. seizure threshold paradigm in mice.

Nitric oxide signaling pathway in the anti-convulsant effect of adenosine against pentylenetetrazol-induced seizure threshold in mice.

The present study was performed to examine the involvement of nitric oxide (NO) signaling pathway in the anti-convulsant effect of adenosine against pentylenetetrazol seizure threshold in mice. Minimal dose of pentylenetetrazol (i.v., mg/kg) needed to induce different phases (myoclonic jerks, generalized clonus and tonic extension) of convulsions was recorded as an index of seizure threshold. Adenosine (100 or 200 mg/kg i.p.) produced a significant increase in the seizure threshold for convulsions induced by pentylenetetrazol i.v. infusion. The anti-convulsant effect of adenosine (100 mg/kg i.p.) was prevented by either L-arginine (50 mg/kg i.p.) [substrate for nitric oxide synthase (NOS)] or sodium nitroprusside (3 mg/kg i.p.) [a NO donor]. On the other hand, N(G)-nitro-L-arginine methyl ester (L-NAME, 2.5 mg/kg i.p.) [a non-selective NOS inhibitor] or 7-nitroindazole (7-NI) (25 mg/kg i.p.) [a specific neuronal nitric oxide synthase (nNOS) inhibitor] potentiated the anti-convulsant action of sub-effective dose of adenosine (50 mg/kg i.p.). Aminoguanidine (100 mg/kg i.p.) [a specific inducible NOS (iNOS) inhibitor] pre-treatment was not effective in inducing anti-convulsant effect with sub-effective dose of adenosine (50 mg/kg i.p.). Furthermore, the increase in seizure threshold elicited by adenosine (100 mg/kg i.p.) was also inhibited by concomitant administration with sildenafil (5 mg/kg i.p.) [phosphodiesterase 5 inhibitor]. In contrast, treatment of mice with methylene blue (1 mg/kg i.p.) [a direct inhibitor of both nitric oxide synthase (NOS) and soluble guanylate cyclase (sGC)] failed to induce anti-convulsant action with adenosine (50 mg/kg i.p.) against pentylenetetrazol i.v. infusion. The results demonstrated that the anti-convulsant action of adenosine in the pentylenetetrazol i.v. seizure threshold paradigm may possibly involve an interaction with the L-arginine-NO-cGMP pathway which may be secondary to the activation of adenosine receptors.

Effect of various antiepileptic drugs in a pentylenetetrazol-induced seizure model in mice.

The present study was undertaken to compare the anticonvulsant effect of various antiepileptic drugs on the intravenous pentylenetetrazol (PTZ)-induced seizure threshold in mice. Minimal doses of PTZ needed to induce different phases (myoclonic jerks, generalized clonus and tonic extensor) of convulsions were recorded as an index of seizure threshold. Furthermore, TID50 (the dose of an anticonvulsant drug required to increase the PTZ seizure threshold for tonic extensor by 50%) was calculated for all drugs, and from these values the potency ratio was determined. Pentobarbital (10-40 mg/kg i.p.), phenobarbital (5-20 mg/kg i.p.), phenytoin (20-40 mg/kg i.p.), carbamazepine (5-20 mg/kg i.p.), diazepam (0.5-2 mg/kg i.p.), chlordiazepoxide (1-4 mg/kg i.p.), triazolam (0.02-0.08 mg/kg i.p.), clonazepam (0.03125-0.25 mg/kg i.p.), GABA (25-100 mg/kg i.p.), ethanol (1000-4000 mg/kg of 10% v/v p.o.), ashwagandha (50-200 mg/kg p.o.), tiagabine (20 and 40 mg/kg i.p.), gabapentin (50-200 mg/kg i.p.), pregabalin (10-40 mg/kg i.p.), progesterone (20-80 mg/kg s.c.), adenosine (25-200 mg/kg i.p.) and rofecoxib (1-4 mg/kg i.p.) exhibited dose-dependent anticonvulsant effects. The TID50 of triazolam was found to be the lowest among all the drugs tested, indicating higher potency. The relative potency of standard drugs to increase the PTZ seizure threshold for tonic extensor was found to be: triazolam > clonazepam > diazepam > rofecoxib > chlordiazepoxide > phenobarbital > carbamazepine > pentobarbital > pregabalin > phenytoin > progesterone > tiagabine > GABA > adenosine > gabapentin > ashwagandha > ethanol. The results of the present study indicate that the intravenous PTZ seizure threshold may be useful for assessing the anticonvulsant effect of drugs effective against different stages of convulsions.

Tiagabine, a GABA uptake inhibitor, attenuates 3-nitropropionic acid-induced alterations in various behavioral and biochemical parameters in rats

Huntingtons disease is an incurable, adult-onset, dominantly inherited neurodegenerative disease. The clinical symptoms of the disease are primarily related to the progressive death of medium spiny gamma-amino butyric acid (GABAergic) neurons in the striatum and the deep layers of the cortex. Further in the later stage of life, the degeneration extends to a variety of brain regions, including the hypothalamus and hippocampus. Various GABAergic agents are being attempted for the treatment of Huntingtons disease. Tiagabine [(R)-N-(4, 4-di-(3-methylthien-2-yl) but-3-enyl) nipecotic acid], a GABA uptake inhibitor, widely used in the treatment of seizures, is suggested to have neuroprotective properties. However, none of the study has elucidated its effect in the treatment of Huntingtons disease and related pathologies. We explored whether tiagabine may attenuate various behavioral and biochemical alterations induced by systemic administration of 3-nitropropionic acid (an inhibitor of complex II of the electron transport chain), an accepted experimental animal model of Huntingtons disease phenotype. Intraperitoneal administration of 3-nitropropionic acid (20 mg/kg., i.p.) for 4 days produced hypolocomotion, muscle incoordination and memory deficit. Daily treatment with tiagabine (5 and 10 mg/kg., i.p.) 30 min prior to 3-nitropropionic acid administration for a total of 4 days, significantly improved the 3-nitropropionic acid-induced motor and cognitive impairment. Biochemical analysis of the whole brain revealed that systemic 3-nitropropionic acid administration significantly increased lipid peroxidation, nitrite levels, total RNA levels and decreased reduced glutathione and succinate dehydrogenase activity which was reversed by daily treatment with tiagabine. Further, there was a decrease in adrenal ascorbic acid levels following daily administration of 3-nitropropionic acid, which was reversed by administration of tiagabine. The results of the present study indicate that tiagabine (5 and 10 mg/kg., i.p.) significantly reversed 3-nitropropionic acid-induced alterations in various behavioral and biochemical parameters and it could be a therapeutic agent for the treatment of Huntingtons disease.

Development and validation of an RP‐HPLC method for the estimation of adenosine and related purines in brain tissues of rats

A new, rapid and sensitive RP-HPLC method with UV spectrophotometric detection was developed and validated for the concomitant estimation of adenosine and related purines in rat brain tissue preparations. The HPLC system consisted of C-18 column with UV-photodiode-array detection ranging from 210 to 400 nm, facilitating the online confirmation of peak purity. The column temperature was maintained at 30 degrees C and the injection volume was 20 muL. Elution with an isocratic mobile phase consisting of water/methanol/acetonitrile (88:5:7 by volume) at a flow rate of 0.8 mL/min yielded sharp, utmost-resolved peaks of adenosine (Ade), inosine (Ino), hypoxanthine (Hypoxan) and adenine (Adn) within 10 min. The method was validated with respect to the linearity, accuracy, precision, sensitivity, selectivity and stability. The method was also employed to estimate the naturally occurring purines in discrete regions of rat brain. A new protocol developed for tissue preparation utilizing H(2)SO(4) and Tris buffer gave well-resolved peaks and high component recoveries (>96%) which eliminated the need of an internal standard. The results show that the method for the determination of Ade, Ino, Hypoxan and Adn by RP-HPLC described here has good linearity, accuracy, precision, sensitivity, selectivity and is simple and rapid to perform.

Effect of systemic administration of adenosine on brain adenosine levels in pentylenetetrazol-induced seizure threshold in mice

Adenosine is one of the inhibitory neuromodulators in the brain. The present study was carried out to elucidate the effect of adenosine on the pentylenetetrazol (PTZ)-induced seizure threshold in mice. Further, the study also correlated the brain adenosine levels in PTZ-induced seizure threshold. PTZ (0.5%, w/v) was infused through lateral tail vein of mouse at a constant rate of 0.3 ml/min until various stages of convulsions were observed. Minimal dose of PTZ (mg/kg) needed to induce different phases (myoclonic jerks, generalized clonus and tonic extensor) of PTZ convulsions were noted as an index of seizure threshold. Intravenous infusion of PTZ resulted in a significant decrease in brain adenosine levels. Systemic administration of adenosine (100 and 200 mg/kg, i.p.), 30 min before PTZ infusion, produced a dose-dependent elevation of PTZ-seizure threshold and also enhanced brain adenosine levels as compared to vehicle treated group. The behavioral and neurochemical observations demonstrated a relationship between adenosine levels in the brain and the PTZ seizure threshold in mice.

Evaluation of Antidepressant-Like Activity of Novel Water-Soluble Curcumin Formulations and St. John's Wort in Behavioral Paradigms of Despair

Background: Curcumin is the active principle of Curcuma longa, one of the widely used components in the traditional system of medicine in India. Despite its efficacy in experimental studies aiming at neuronal disorders like depression, curcu-mins poor water solubility challenges the production of therapeutic formulations. This study investigates the antidepressant-like activity of novel water-soluble curcumin formulations, dispensed in three different concentrations. Further, the study comparatively evaluates St. Johns wort (SJW), another herbal preparation. Methods: These compounds were evaluated in the forced swimming test in mice, and the corresponding changes in the neurotransmitter levels were measured. Results: Three water-soluble curcumin formulations, C-5, C-20 and C-50 (50-200 mg/kg p.o.) decreased the immobility period, and increased serotonin and dopamine levels in the brain tissues. A subeffective dose (50 mg/kg) of these formulations enhanced the antidepressant-like effect of classical antidepressants with varied mechanisms of action. In addition, an SJW dose of 25 mg/kg showed a significant antidepressant-like effect in all the behavioral studies and also significantly increased brain neurotransmitter levels, especially that of serotonin. The effects produced by C-5 were comparable with those of SJW and fluoxetine, respectively. Conclusion: In all these observations, the water-soluble formulations showed a significant antidepressant-like effect, including enhancement of neurotransmitter levels as compared to the similar dose of a conventional curcumin preparation. Thus, these formulations may be used as a novel treatment option in the management of mental depression.

Systemic administration of adenosine ameliorates pentylenetetrazol-induced chemical kindling and secondary behavioural and biochemical changes in mice

Adenosine is one of the inhibitory neuromodulators in the brain and is considered to be responsible for seizure arrest and postictal refractoriness. Adenosine, adenosine receptor agonists, and adenosine uptake blockers are known to reduce the severity and duration of amygdala‐kindled seizures. The present study was carried out to elucidate the anticonvulsant and neuromodulatory effect of systemic adenosine on the pentylenetetrazol (PTZ)‐induced chemical kindling in mice. Kindling was induced by chronic administration of a subconvulsive dose of PTZ (40 mg/kg, i.p.) on every other day for a total period of 9 days. Adenosine was administered daily, 30 min before PTZ or vehicle. The kindling score was recorded immediately following PTZ administration according to a prevalidated scoring scale. Various behavioral and biochemical estimations were performed on day 10 (i.e. 24 h after the last dose of PTZ). Chronic PTZ treatment progressively increased the seizure score with the maximum score reached on day 9. Behavioral analysis found hyperlocomotor activity, anxiogenic response, hyperalgesia and amnesia in kindled mice. Biochemical analysis revealed that chronic treatment with PTZ significantly increased lipid peroxidation (malondialdehyde levels), nitrite (NO2− levels), adenosine deaminase (ADA) and total RNA levels and decreased catalase, reduced glutathione (GSH) levels in brain homogenates, and a depletion of adrenal ascorbic acid. Daily treatment with adenosine (25 and 50 mg/kg, i.p.) for 9 days led to a significant decrease in PTZ‐induced kindling score and also reversed various behavioral and biochemical alterations produced by PTZ. The results of the present study suggested that systemic adenosine administration reversed the behavioral and biochemical alterations induced by chronic PTZ.

Estimation of adenosine and its major metabolites in brain tissues of rats using high-performance thin-layer chromatography-densitometry.

A new, rapid and sensitive high-performance thin-layer chromatographic (HPTLC) method with densitometry was developed and validated for the concomitant estimation of purines like adenosine (Ade) and its major metabolites, inosine (Ino) and hypoxanthine (Hypoxan) in rat brain tissue preparations. The HPTLC method was chosen in order to generate better resolution and evade the tedious and prolonged sample preparation methods necessarily performed with HPLC methods when analyzing biological samples. In this method the planar chromatography was executed on aluminum plates pre-coated with silica gel 60 F(254). Elution was performed with a two-step gradient mobile phase consisting of solvent A [n-butanol/water/acetonitrile/10% ammonia solution/glacial acetic acid (5:2:4:1:0.5, v/v)] and solvent B [n-butanol/chloroform/acetonitrile/10% ammonia solution/glacial acetic acid (5:4:2:1:0.5, v/v)]. The quantitative analysis of purines was performed based on the peak areas obtained from the reflectance scanning densitometry, performed at 258nm. The spectral scan was done from 200 to 300nm which facilitated the spectral analysis of peaks for purity and spectral matching. The method was validated in terms of linearity, accuracy, precision, sensitivity, selectivity and repeatability. The method was successfully employed to estimate the endogenous purines in discrete regions of rat brain. A novel protocol developed for the tissue preparation utilizing 0.1M HCl and 0.15M NaOH solutions made in 60% (v/v) methanol resulted in well-resolved peaks and high component recoveries. The results for the first time show that this method established for the flexible estimation of Ade, Ino and Hypoxan by planar chromatography has good linearity, accuracy, precision, sensitivity, selectivity and is simple, rapid and moreover, economical to produce maximum resolution in brain tissue preparations.