Low-intensity transcranial ultrasound and Mucuna Pruriens for treatment of Parkinson’s disease in mice

다국어 초록 (Multilingual Abstract)

Parkinson’s disease (PD) is the second leading neurodegenerative disease after Alzheimer’s disease. It is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta of the brain and aggregation of the alpha synuclein, resulting in motor, non-motor, and olfactory impairment of the body. Mucuna Pruriens (MP) is a nutraceutical product which contains Levodopa (L-DOPA) and has proven effects in improving the symptoms of PD. Moreover, low-intensity transcranial ultrasound (LITUS) can activate neurons to release dopamine and help in alleviating the symptoms of PD.
In this study, we investigated the anti-parkinsonian potential of MP and LITUS against PD. We first developed an in vivo model of the PD in C57BL/6 male mice using rotenone. The number of mice used in this experiment was 32. There were 2 phases of mice experiment. The 1st phase of the experiment was performed to investigate MP effects on PD mice compared with the L-DOPA. These experiments were performed to validate the effectiveness of MP to treat PD in relation to standard treatment of PD. For that purpose, synthetic L-DOPA in a ratio of 1:20 with MP was used as it contains 5% L-DOPA by weight in it. 12 mice were used in this phase and were divided into 4 groups of 3 mice such as control group, PD group, PD treated with MP and PD treated with L-DOPA. The 2nd phase was performed to investigate the effect of MP and LITUS individually and collectively on PD mice. 20 mice were divided into 5 groups of 4 mice which were control, PD, PD with MP, PD treated with LITUS and PD treated with both MP and LITUS groups. The PD was induced in 28 days while PD treatment lasted for 19 and 10 days in the 1st and 2nd phases, respectively. Beam balance test, olfactory test, and serum enzyme-linked immunosorbent assay (ELISA) analysis test for IL-12, IL-6, TGF-β1 was performed to validate the PD inducement and treatment.
The levels of IL-12, IL-6 and TGF-β1 (p<0.0001) in PD mice model group were significantly higher than those in the control group. The PD mice also showed higher latencies in beam balance and olfactory tests (p<0.0001) as compared to the control group. In the 1st phase, both MP and L-DOPA treated groups showed alleviation in latencies in beam balance and olfactory tests and decreased neuroinflammation in ELISA analysis (p<0.001). The results treated by MP and L-DOPA showed insignificant difference in their values (p>0.05). This proved that the MP and L-DOPA have similar effects in improving the symptoms of PD when used in the ratio of 1:20. In the 2nd phase, after being treated with MP the level of IL-6 (p<0.05), TGF-β1 (p<0.001) and IL-12 (p<0.0001) were significantly reduced as compared to untreated PD mice. While treatment with LITUS also downregulated the levels of IL-6 (p<0.01), TGF- β1 (p<0.001) and IL-12 (p<0.01) as compared to untreated PD mice. The PD mice treated by the MP+LITUS showed less latency (p<0.05) and less time (p<0.001) as compared to untreated PD mice during beam balance and olfactory tests, respectively. These results proved the effectiveness of both MP and LITUS in reducing the level of neuroinflammation and improving the behavioral symptoms in PD mice model. Moreover, PD mice being treated with MP and LITUS collectively showed better results in olfactory test but not for other factors as compared to untreated PD mice. These findings suggest that each MP and LITUS may have a potential therapeutic effect on PD but not much synergically by modulating the inflammatory response of the body.
Furthermore, both MP and LITUS reduced the level of IL-6 and TGF-β1 in this study. It may be inferred that reduction in the level of IL-6 and TGF-β1 eventually leads to the reduction of the Th17 cells. The pathogenic Th17 is thought to be present in virtually all chronic inflammatory disorders. This can be an interesting area of research in further understanding the immunological effect of MP and LITUS in ameliorating the PD symptoms. 

목차 (Table of Contents)

• ABSTRACT １
• 1. INTRODUCTION ３
• 1.1 PARKINSON'S DISEASE ３
• 1.1.1 BASIS OF THE PARKINSON'S DISEASE ３
• 1.1.2 CURRENT TREATMENTS ５
• 1.1.3 LIMITATION OF THE CURRENT TREATMENTS ５
• 1.2 ROLE OF NEUROINFLAMMATION IN PARKINSON'S DISEASE ６
• 1.3 ULTRASOUND ８
• 1.3.1 THERAPEUTIC ULTRASOUND ８
• 1.3.2 LOW-INTENSITYT TRANSCRANIAL ULTRASOUND AND NEUROINFLAMMATION ９
• 1.4 ROLE OF MUCUNA PRURIENS IN CURING PARKINSON'S DISEASE １０
• 1.5 OBJECTIVE AND ORIGINALITY １１
• 2. METHODS １３
• 2.1. INDUCTION OF PARKINSON DISEASE IN MICE １３
• 2.2. SYNTHETIC DRUG L-DOPA TO USE AS A POSITIVE CONTROL DRUG １５
• 2.3. PARAMETERS TO VALIDATE PARKINSON'S DISEASE AND TREATMENT １５
• 2.4 EXPERIMENTAL PLAN １９
• 2.5 MUCUNA PRURIENS (MP) AND PREPARATION OF MP WATER EXTRACT ２１
• 2.6 ULTRASOUND NEUROSTIMULATION ２２
• 3. RESULTS ２４
• 3.1 PARKINSON'S DISEASE INDUCTION 2４
• 3.2 COMPARATIVE STUDY OF MP AND L-DOPA IN IMPROVING THE PHYSICAL AND INFLAMMATORY SYMPTOMS OF PARKINSON'S DISEASE ２７
• 3.2.1 EFFECTIVENESS OF MP IN IMPROVING THE SYMPTOMS OF PD IN COMPARISON TO L-DOPA ２７
• 3.3 EFFECTIVENESS OF MUCUNA PRURIENS AND LITUS TO TREAT PHYSICAL SYMPTOMS OF PARKINSON'S DISEASE ３０
• 3.4 EFFECTIVENESS OF MUCUNA PRURIENS AND LITUS TO ALLEVIATE SERUM BIOMARKER LEVELS OF PARKINSON'S DISEASE ３０
• 3.5 EFFECTIVENESS OF MP IN IMPROVING SYMPTOMS OF PARKINSON'S DISEASE ENHANCED BY LOWINTENSITY THERAPEUTIC ULTRASOUND ３１
• 4 DISCUSSION ３５
• 5 CONCLUSION ３９
• APPENDIX ４０
• EXPERIMENTAL PROTOCOL: ４４
• DESCRIPTION OF THE METHODOLOGIES/EXPERIMENTAL DESIGN:. ４５
• REFERENCES ４９