STUDY OF THE ACTIVITY OF BREADFRUIT (ARTOCARPUS ALTILIS [PARK.] FOSBERG.) EXTRACTS ON MULTIPLE SCLEROSIS RAT MODEL

• Main Author: Hadi Setya Palupi, Dwi
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/54726
• Institution: Institut Teknologi Bandung

The pathology and symptoms of multiple sclerosis (MS) are very complex and vary from person to person. Currently incurable, and there is still no appropriate and effective therapy for MS to inhibit the neurodegenerative rate. Anti-inflammatory, antioxidant, immunosuppressant, and neuroprotective are required to reduce symptoms and prevent the rate of neurodegeneration. Symptomatic treatment approaches of MS can use herbal medicine as an alternative because herbal medicine has the advantage of multi-targeted therapy. The leaves, fruit, bark, and roots of the breadfruit (Artocarpus altilis [Park.] Fosberg) contain many bioactive compounds with activities thought to have the potential to reduce MS symptoms. Experimental autoimmune encephalomyelitis (EAE) was an animal model of MS frequently used in research. MS with repeated attacks (relapse-remission) is common in humans, but the induction of EAE in animals more often results in a chronic progressive model. The inductor is one factor that can be modified to form an animal model as close as possible to MS in humans. This study aimed to examine the pharmacological activity of the ethanol extract of several parts of breadfruit plants that have the potential for MS therapy; to optimize the EAE induction method by using several inductor combinations that produce clinical and demyelinating patterns as closely as possible with relapse-remission MS; to study the effectiveness of selected extracts on the symptom pattern and remyelination of EAE rats. The study was divided into four stages: preparation of the test material, screening of the extract's pharmacological activity, optimising the EAE induction method and assessing the effectiveness of breadfruit plant extracts in EAE rats. The extract of leaves (EDS), stem bark (EKBS), and whole fruit (EBS) of breadfruit were obtained by maceration using 96% ethanol as a solvent. The yields of EDS, EKBS, and EBS were 8.28%, 4.62% and 13.87%, respectively. In phytochemical screening showed the presence of flavonoids and steroids/triterpenoids in all three extracts. Screening for pharmacological activity includes in vitro and in vivo antiinflammatory, antioxidant, immunosuppressant and neuroprotective assay. The results of the anti-inflammatory test showed that EDS 200 mg/kg bw could significantly (p <0.05) reduce Complete Freund's Adjuvant (CFA)-induced chronic inflammation with the highest percentage of inflammation inhibition (43.23%) was achieved on day 15. In vitro antioxidant activity was determined through a scavenging activity assay against 2,2-Diphenyl-1- Picrylhydrazyl (DPPH) and nitric oxide (NO) radicals. EDS and EBS showed strong DPPH radical scavenging activity with IC50 values of 72.17±0.84 and 66.34±1.70 µg/mL. The most potent scavenging activity of NO radicals was shown by EDS with an IC50 value of 45.40 ± 6.49 µg/mL. The results of the ex vivo antioxidant activity test in CFA-induced chronic inflammation rats showed that EDS 200 mg/kg bw significantly reduced (p <0.01) H2O2 levels (36.21%) and increased SOD activity (75.72%). The results of the immunosuppressant activity test showed that the EDS 200 mg/kg BW significantly reduced (p <0.01) the phagocytic index of the reticuloendothelial system, suppressed the cellular and humoral immune response, which was marked by a significant decrease in inflammation in the delayed-type hypersensitivity reaction at 24 and 48 hours, and lower primary antibody titres than controls. The neuroprotective activity test in vitro showed that the three extracts had moderate potential as acetylcholinesterase inhibitors. The results of in vivo testing with the water maze method (Morris water maze) showed that administering EDS mg/kg bw for seven days improved the learning ability of mice and significantly increased (p <0.05) the spatial memory of mice scopolamine hydrobromide-induced amnesia. The part of the breadfruit plant with four activities, anti-inflammatory, antioxidant, immunosuppressant and neuroprotective, was the leaves. EDS at a dose of 200 mg/kg bw was the selected extract to be tested on animal models of MS because it was more potent than other extracts. Optimization of the EAE induction method was carried out by comparing several inductor combinations: (1) Cuprizone, encephalitogen, and Bordetella pertussis suspension; (2) Cuprizone, ovalbumin, and encephalitogen; (3) encephalitogen and B. pertussis suspension; (4) Ovalbumin and encephalitogen. The results showed that the combination of inductor 1 produces EAE that resembles MS relapseremission in humans. The work indicated the characteristics of the neurological score: incidence of 100%, highest relapse frequency (three times), a highest neurological score of 2.00, 16.67% mortality. CNS damage was characterized by focal demyelination in white and grey matter, perivascular cuff expansion, inflammation, and gliosis. Microglia reactivity in the cerebrum and serum TNF-? levels increased significantly (p<0.01) compared to the normal group. Combination 1 was used as an inductor for the EAE induction method at the stage of assessing the effectiveness of breadfruit plant extracts in EAE rats. The administration of EDS 200 mg/kg bw for 14 days significantly reduced (p <0.05) the cumulative score and frequency of relapses and the severity of neurological symptoms was 17.18%; improve inflammation characterized by improvement in brain histology and decreased expression of the gfap gene; significantly (p<0.05) reduced NO radical levels and increased catalase activity compared to the EAE group. EDS 200 mg/kg bw also increases remyelination which is indicated by an increase in olig2 gene expression and myelin thickness. In summary, breadfruit leaf extracts at a dose of 200 mg/kg bw had potent antiinflammatory, antioxidant, immunosuppressant and neuroprotective activities. A combination of cuprizone, encephalitogen and B. pertussis suspension produced an EAE rat model that resembled MS relapse remission in humans. Breadfruit leaf extract at 200 mg/kg bw reduced the severity of clinical/neurological symptoms, inflammation, oxidative stress, and increased remyelination in EAE rats.