Phytochemicals are considered as natural bio-active compounds with extraordinary bio-activities like free radical scavenging, enhancing mitochondrial integrity. preventing severe inflammation, regulating apoptosis and inhibiting toxic protein aggregations. This presentation deals with how phytochemicals are promoting brain health against various molecular assaults and wide range of diseases and disorders.

1. Natural Bioactive Compounds
Promotes Neurohealth
Dr. D. VIJAYRAJA
HEAD
DEPARTMENT OF BIOCHEMISTRY
Rev. JACOB MEMORIAL CHRISTIAN COLLEGE
AMBILIKKAI-624612
ODDANCHATRAM
ONE DAY VIRTUAL WEBINAR
23.03.21

2. The Brain

3. Protecting Brain Health
OVERALL WELLBEING MAY HELP TO
MAINTAIN GOOD BRAIN HEALTH.
Strive for:
Healthy
eating
Regular
exercise
Keeping
your brain
active
Social
connections
Getting
enough
sleep

4. PHYTOCHEMICALS FOR BRAIN
• Project Description
• Project Methodology
• Key Findings/Results
• Research drilldown
• Conclusion
A healthy diet may promote brain health
now, and in the years to come.

5. Possible Risks or Threats
to Brain Health
• Some medicines, or improper use of them
• Smoking
• Excessive use of alcohol
• Heart disease, diabetes, and other health problems
• Poor diet
• Insufficient sleep
• Lack of physical activity
• Little social activity and being alone most of the time
• Environmental toxins

6. Dietary phytochemicals and their action against
oxidative stress and human diseases
The multi-target effects of phytochemicals in the
brain

7. Myricetin and its Neuroprotective effect
Yellow crystals of myricetin 1HNMR Spectrum of isolated flavonol myricetin from
Turbinaria ornata

8. Parkinson’s Disease

9. Etiology of Parkinson’s disease
Dopaminergic pathways in Brain Region of brain showing dopaminergic
neuron density in healthy and PD subjects

10. Arup Kumar Bishoyi, NIPER
Pathophysiology in PD

11. Effect of myricetin on intracellular ROS production
Preventive effect of myricetin against rotenone induced ROS generation by DCFDA staining and
its Densitometry. (2,7-diacetyl dichlorofluorescin diacetate-DCF DHA)
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and
#p < 0.05 compared to rotenone group (DMRT).

12. The effect of myricetin on rotenone induced ROS in
SK-N-SH cells : TBARS and GSH Levels
(A and B): Levels of TBARS and GSH in experimental groups.
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).

13. The effect of myricetin on rotenone induced ROS in
SK-N-SH cells : SOD, CAT’ and GPx activities 13
Figures (C, D and E): Influences of myricetin on the activities of against rotenone toxicity enzymatic
antioxidants.
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group
(DMRT). AEnzyme concentration required for 50% inhibition of nitroblue tetrazolium reduction in 1 min. BMicromoles of hydrogen
peroxide consumed per minute. CMicrograms of glutathione consumed per minute.

14. Effect of myricetin on mitochondrial membrane potential
(ΔΨm)
Alterations in mitochondria membrane potential measured by rhodamine 123 staining. and
respective Densitometry. (Rhodamine 1,2,3)
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and
#p < 0.05 compared to rotenone group (DMRT).
Control Rotenone Myricetin
+Rotenone
Myricetin

15. Effect of myricetin on Apoptosis
Apoptotic morphological changes in control, rotenone, myricetin pre-treated and rotenone and
myricetin alone treated SK-N-SH cells and their respective Densitometry. (EtBr, Acridine orange)
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).
Control Rotenone Myricetin
+Rotenone
Myricetin

16. Effect of myricetin on rotenone induced cyt c and
caspases 3 and 9 expressions in SK-N-SH cells
Expressions of Bax, Bcl-2, in control and experimental groups. Protein expressions were
quantified by using β-actin as an internal control. The band density was quantified by scanning
densitometry.
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).

17. Effect of myricetin on rotenone induced cyt c and
caspases 3 and 9 expressions in SK-N-SH cells
Expressions of Caspases-3, -9 and cyt-c in control and experimental groups. Protein
expressions were quantified by using β-actin as an internal control. The band density was
quantified by scanning densitometry.
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).

19. Rotenone lethal dose study (LD 50) and myricetin
effective dose fixation
Effective dose assessment of rotenone (250, 500, 750 and 1000μM) and myricetin (0.025%,
0.05%, 0.1% and 0.2%) by mortality %. 50% of mortality (LD50) is obtained at 500 μM and
protective dose to reduce mortality was fixed as 0.1% in rotenone and myricetin respectively.
Values are presented as mean ± SD of four experiments in each group.

20. Behavioural assessment studies:
Open field test and Climbing assay
Openfield test:The time taken in Open field test (Number of squares crossed) for exploring,
acclimatization and basal locomotor activity. Climbing assay: The time taken to climb 25 cm long
glass tube in control and experimental groups in 60s of climbing assay.
Values are given as mean  S.D for 3 vials of drosophila 50 flies each group. Values not sharing a common superscript lesymbol
differs significantly at p<0.05 (DMRT).

21. Styrofoam bead holding, bead maintenance, bead rotation and bead slipped by experimental group
flies
Values not sharing a common superscript letter differed significantly at p<0.05 (DMRT).
Behavioural assessment studies:
Styrofoam bead test of experimental groups

22. Foot-print test for walking pattern and gait analysis
Foot print test for gait assessments
Control Rotenone
Myricetin
+Rotenone
Myricetin

23. T maze assay: Analysng the learning ability and short
term memory of experimental group flies
T-Maze apparatus and T-Maze aversive phototaxis suppression assay for learning ability and short
term memory assay. Values not sharing a common superscript letter differ significantly at p<0.05 (DMRT)
A

24. The effect of myricetin on rotenone induced ROS in
Drosophila melanogaster : TBARS and GSH Levels
Levels of TBARS and GSH in experimental groups.
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and
#p < 0.05 compared to rotenone group (DMRT).

25. The effect of myricetin on rotenone induced ROS in
experimental groups: SOD, CAT’ and GPx activities
Influences of myricetin on the activities of enzymatic antioxidants against rotenone toxicity .
Values are presented as mean ± SD of each group. *p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).
AEnzyme concentration required for 50% inhibition of nitroblue tetrazolium reduction in 1 min. BMicromoles of hydrogen peroxide
consumed per minute. CMicrograms of glutathione consumed per minute.

26. Effect of myricetin on MAO B activity in control and
experimental groups.
Effect of myricetin on MAO B activity in control and experimental groups.
Values are given as mean ± S.D for 3 vials of drosophila 50 flies each group. Values not sharing a
common superscript letter differ significantly at p<0.05 (DMRT).

27. Effect of Myricetin on rotenone induced
dopaminergic neuron clusters degeneration and
dopamine level in experimental groups.
Tyrosine hydroxylase immunolabeling showing dopaminergic neuron clusters in brains of experimental
groups. Dopamine levels in the control and experimental groups. Values are given as mean ± S.D for 3 vials of
drosophila 50 flies each group. Values not sharing a common superscript letter differ significantly at p<0.05 (DMRT).

28. Effect of Myricetin on rotenone induced inflammation
in experimental groups.
Effect of myricetin on rotenone induced pro-inflammatory and anti-inflammatory cytokines expression
in experimental group of flies and respective densitometry. Values are given as mean ± S.D for 3 vials of drosophila
50 flies each group. Values not sharing a common superscript letter differ significantly at p< 0.05 (DMRT).

29. Effect of Myricetin on rotenone induced apoptosis in
experimental groups.
Effect of myricetin on rotenone induced changes in the expressions of apoptotic protein markers (Bax, Bcl-2,
caspase 3, 9 and cyto-c) in control and experimental flies. Protein expressions by
using β-actin as an internal control. Values are given as mean ± S.D for 3 vials of drosophila 50 flies each group. Values
not sharing a common superscript letter differ significantly at p< 0.05 (DMRT).

30. Conclusion
Our study results show that myricetin prevents the onset of adverse changes in PD
progression in cell line and inhibits DA death, promotes the muscular coordination and
psychological well being of fruit flies.
Thus the amelioration of undesirable changes, apoptosis in dopaminergic neurons, their
degeneration and maintenance of dopamine level clearly elucidates the neuroprotective
activity of myricetin and its significant role in prevention of PD.
Recommendations
Myricetin may be used as a drug molecule in treating Parkinson’s disease either alone, or
through diet and along with the existing treatment methods.
Future prospecting of myricetin to treat Parkinson’s disease through investigating its
neuroprotective effect in higher animal models and in human trials.

31. Significant Phytochemicals
in our daily food

32. 32
Glimpse of our research activities in our Experimental Neuroscience Lab

33. THANKS!