Pharmacological Evaluation of C-Phycocyanin Against Cuprizone Induced Multiple Sclerosis Mice Model

Abstract

Background Multiple sclerosis is an autoimmune disorder. It is a chronic inflammatory demyelinating disorder of the central nervous system depicted by changes in motor and sensory functions and disruption in neuronal transmission causing loss in neuronal functions and cognitive functions leading to inflammation and axonal degeneration. The conventional therapies for MS are mainly symptomatic and focus on reducing the severity of the disease or slowing the progression of the disease which primarily include disease modifying therapies. This data depicts the urgent need for development of alternative therapy for the therapy of multiple sclerosis. The drug C- phycocyanin is obtained from blue-green microalgae Spirulina. It is reported to possess antioxidant, anti-inflammatory and remyelinating activity and also cause regeneration of white matter. Objectives: In the light of existing reports, this study was undertaken to evaluate the pharmacological action and to investigate the mechanism of action of C-phycocyanin in Cuprizone intoxicated mice model of multiple sclerosis. Materials & Methods: The Balb/c mice were separated into 6 groups on basis of equal weight and sex. All the animal groups except Normal Control were given cuprizone toxin at dose of 400 mg/kg orally for 6 weeks. The treatment was started from 3rd week; standard treatment group was administered with Methylprednisolone in the dose of 10 mg/kg for two weeks and 5mg/kg for the two last week; US FDA states that by giving higher dose and then reducing the dose has proven more effective in reducing acute lesions in MS. The first treatment group [D+T1] received test drug C-phycocyanin in low dose of 100 mg/kg, second treatment group [D+T2] received medium dose of C-Phycocyanin i.e., 250 mg/kg and the third treatment group received high dose C-Phycocyanin of 500 mg/kg. The neurobehavioral parameters like Rota rod, Tail flick, Morris water maze and Y-maze were performed. Biochemical parameters like Oxidative stress [SOD & Catalase activity estimation], Nitrosative stress parameters [NO level estimation] and neuroinflammatory [Il-6, TNF-α & NF-kappa B estimation] and neuroprotective parameters [β-NGF & Neprilysin] were performed. Also, histological and immunohistochemistry assessment was carried out. Result In this study, multiple sclerosis is induced by cuprizone toxin, which causes demyelination, loss of neuronal functions and also loss of locomotor coordination and balance, together with memory loss, muscle impairment etc. The administration of C-Phycocyanin has shown improvement in motor coordination as seen by the results of Rota rod. The results of Morris water maze and Y maze shows that C-Phycocyanin improves the working memory and spatial learning ability in mice. Acute nociceptive response is also improved by the drug which shows remyelination and improved nerve response. Super oxide dismutase activity is a marker for the oxidative stress. During inflammation, there is significant increase in oxidative stress thus the SOD levels are decreased. Higher SOD levels indicated the lower inflammation and thus this depicts that the treatment of C-phycocyanin is effective as anti-oxidant agent. In inflammatory disease there is a rise in nitric oxide levels. There is significant increase in disease control group when compared to normal control and treatment groups. Medium treatment receiving group and standard drug treatment group shows the significant decrease in nitric oxide levels. Also, neuroinflammatory parameters like IL-6 and NF-kappa B shows reduced inflammation compared to DC groups which shows anti-inflammatory property of C-Phycocyanin. Lastly, results of β-NGF and Neprilysin shows neuroprotective effect of the drug.

Conclusion Our data suggest that C-Phycocyanin has anti-inflammatory and anti-oxidant activity and also contributes to promotion of neuronal health by providing neuroprotective effect against cuprizone induced Multiple sclerosis in mice. It has appropriate clinical application given that demyelination and neurodegeneration are main indication in the patients of MS, connected with oxidative injury, meningeal inflammation and axonal loss.