THE AMELIORATING EFFECT OF AQUEOUS EXTRACT BACOPA MONNIERI

ON THE PREFRONTAL CORTEX IN PROPIONIC ACID-INDUCED AUTISM IN

MALE ALBINO WISTAR RAT MODEL

TABLE OF CONTENTS

Title page
Certification page
Dedication
Acknowledgement
Abstract
Table of contents
List of figures
List of plates

CHAPTER ONE: INTRODUCTION

1.1 Background of study
1.2 Aim and Objective
1.3 Significance of study
1.4 Scope of study

CHAPTER TWO: LITERATURE REVIEW

2.1 Review of relevant literature
2.2 Herb of Study
2.2.1 Phytochemical constituent of Bacopa monnieri
2.2.2 Uses of Bacopa monnieri
2.3 Organ of study
2.3.1 Gross anatomy of prefrontal cortex
2.3.2 Histology of prefrontal cortex
2.3.3 Embryology of prefrontal cortex

CHAPTER THREE: MATERIALS AND METHOD

3.1 Ethical approval
3.2 Breeding of animals
3.3 Preparation of aqueous extract of Bacopa monnieri
3.4 Administration of extract
3.5 Experiment protocols
3.6 Termination of experiment
3.7 Tissue processing

CHAPTER FOUR: RESULTS

4.1 Histological observation
4.2 Photomicrographs
CHAPTER FIVE: DISCUSSION, SUMMARY, CONCLUSION AND
RECOMMENDATION
5.1 Discussion
5.2 Summary
5.3 Conclusion
5.4 Recommendation for further studies
5.5 Contrition to knowledge

REFERENCES
CHAPTER ONE

BACKGROUND OF STUDY

Autism spectrum disorder (ASD) is a complex developmental condition involving persistent

challenges with social communication, restricted interests and repetitive behaviour. While
autism is considered a lifelong disorder, the degree of impairment in functioning because of
these challenges varies between individuals with autism (American Psychiatric Association,
n.d.). The prevalence of ASD has increased dramatically over the past few decades, now
affecting approximately 1 in 36 children aged 8 years, according to the Morbidity and
Mortality Weekly Report (Maenner et al., 2023).

The aetiology of autism spectrum disorder (ASD) is complex and involves an interplay of
genetic, epigenetic, and environmental factors. Although the exact causes and mechanisms of
ASD remain unidentified, extensive research has identified various risk factors from both
genetic and environmental sources. Genetic and epigenetic elements influence the expression
of risk genes, leading to a highly variable disease presentation even among those with similar
genetic mutations. Genetic factors include copy number variations (CNVs) and mutations,
while epigenetic factors encompass DNA methylation, microRNAs (miRNAs), and
chromatin remodelling. Additionally, many environmental factors that induce epigenetic
changes in chromatin contribute to an increased risk of ASD. (Khogeer et al., 2022).
Despite significant advancements in understanding ASD, effective treatments remain limited,
highlighting the urgent need for novel therapeutic approaches.

While genetic factors significantly contribute to ASD, environmental influences, including

exposure to neurotoxic substances, have been implicated in its development (Lyall et al.,
2014). Propionic acid (PPA), a short-chain fatty acid produced by gut microbiota, has gained
attention for its role in neurodevelopmental disorders. Elevated levels of PPA in the
gastrointestinal tract have been observed in individuals with ASD, suggesting a potential link
between gut microbiota and autism. Although PPA may be beneficial at appropriate levels,
such as improving insulin sensitivity, lowering cholesterol, and reducing food intake,
excessive PPA may have many negative effects on health and behavior. (MacFabe, 2012).

Animal models, particularly rodent models, have been instrumental in studying ASD. The
administration of PPA intracerebroventricularly to rodents has been shown to induce
behaviors and biochemical changes reminiscent of human ASD, such as neuroinflammation,
oxidative stress, and disruptions in neurotransmitter systems (Shultz et al., 2008). These
models are crucial for understanding the pathophysiology of ASD and evaluating potential
therapeutic interventions.

Herbal medicine has been utilised for centuries across various cultures to treat numerous
ailments. Recently, there has been renewed interest in natural products as potential treatments
for neurodevelopmental and neuropsychiatric disorders (WHO, 2023). One such herb,
Bacopa monnieri, known as Brahmi, has gained attention for its cognitive-enhancing and
neuroprotective properties. Traditionally used in Ayurvedic medicine to improve memory
and cognitive function, Bacopa monnieri is being explored for its relevance in addressing
neurological conditions like ASD in which memory is also linked (Kongkeaw et al., 2014;
Singh & Dhawan, 1997). The active compounds in Bacopa monnieri, called bacosides, have
demonstrated antioxidant, anti-inflammatory, and neuroprotective effects (Russo & Borrelli,
2005). Research indicates that Bacopa monnieri can alleviate cognitive deficits and oxidative
stress in various neurological disorders, underscoring its potential as a therapeutic agent for
neurodevelopmental conditions such as ASD (Calabrese et al., 2008).

The prefrontal cortex (PFC) is a critical brain region constituting the highest level of cortical
hierarchy involved in representation of actions, executive functions, social behaviour, and
cognitive processes (Fuster, 2001). Impairments in the PFC have been linked to the core
symptoms of ASD, such as difficulties in social interaction and executive functioning
(Mohapatra & Wagner, 2023). Therefore, targeting the PFC with neuroprotective agents may
help alleviate some ASD symptoms.

Given the potential neuroprotective effects of Bacopa monnieri, it is hypothesised that its
aqueous extract could mitigate the neurotoxic effects of propionic acid (PPA) on the
prefrontal cortex (PFC) and subsequently improve behavioural outcomes in an autism
spectrum disorder (ASD) rat model. Due to the limited literature on the effects of Bacopa
monnieri on the PFC, this study aims to investigate the biochemical, histological, and
behavioural changes resulting from the administration of Bacopa monnieri aqueous extract in
male albino Wistar rats induced with PPA to model autism. This research seeks to provide
insights into the potential of Bacopa monnieri as a therapeutic agent for ASD.

AIM AND OBJECTIVE

This study aims to investigate the potential ameliorating effect of the aqueous extract of
Bacopa monnieri on the prefrontal cortex in propionic acid-induced autism using a male
albino Wistar rat model. Its objective is to assess the histological changes and evaluate the
impact of Bacopa monnieri aqueous extract on mitigating histological alterations in the
prefrontal cortex of propionic acid-induced autism in male albino Wistar rats.

SIGNIFICANCE OF THE STUDY

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with limited

treatment options. Thus, investigating the potential ameliorating effect of Bacopa monnieri
on propionic acid-induced autism has significant implications.

This study provides insight into a potential novel therapeutic approach for ASD. If the
aqueous extract of Bacopa monnieri demonstrates efficacy, it could offer a natural and
alternative treatment option for individuals with autism. Understanding the effect of Bacopa
monnieri on the prefrontal cortex histology can shed light on its neuroprotective potential.
This knowledge could contribute to the development of neuroprotective strategies against
ASD-related neuropathology.
Assessing histological changes in the prefrontal cortex provides valuable insights into the
neuropathological mechanisms underlying ASD and the potential mechanisms of action of
Bacopa monnieri. If the aqueous extract of Bacopa monnieri demonstrates effectiveness in
mitigating histological alterations, it could pave the way for further clinical studies and the
development of Bacopa monnieri-based therapies for ASD. Bacopa monnieri is a widely used
herb with neuroprotective properties. Demonstrating its efficacy in ameliorating ASD-related
changes could support its use in natural medicine approaches for neurological disorders.

Also, if efficacy is demonstrated there will be reduced dependence on expensive medications

and affordable alternatives or complementary therapies will be offered to autistic individuals.
Many pharmaceutical treatments for ASD come with significant side effects, which can lead
to additional healthcare costs. Bacopa monnieri is generally considered to have a favourable
safety profile, potentially reducing the costs related to managing side effects.

In summary, this study has the potential to contribute significantly to the understanding of
ASD pathophysiology and may lead to the development of new therapeutic and affordable
strategies using natural compounds.

SCOPE OF STUDY

This study is focused on the histological examination of prefrontal cortex of the male albino
wistar rat in propionic acid-induced autism using Bacopa monnieri for a specific period of
administration.
REFERENCES

1. American Psychiatric Association. (n.d.). What is autism spectrum disorder?

https://www.psychiatry.org/patients-families/autism/what-is-autism-spectrum-
disorder

2. Maenner, M. J., Warren, Z., Williams, A. R., Amoakohene, E., Bakian, A. V., Bilder,
D. A., Durkin, M. S., Fitzgerald, R. T., Furnier, S. M., Hughes, M. M., Ladd-Acosta,
C. M., McArthur, D., Pas, E. T., Salinas, A., Vehorn, A., Williams, S., Esler, A.,
Grzybowski, A., Hall-Lande, J., Nguyen, R. H. N., … Shaw, K. A. (2023). Prevalence
and Characteristics of Autism Spectrum Disorder Among Children Aged 8 Years -
Autism and Developmental Disabilities Monitoring Network, 11 Sites, United States,
2020. Morbidity and mortality weekly report. Surveillance summaries (Washington,
D.C. : 2002), 72(2), 1–14. https://doi.org/10.15585/mmwr.ss7202a1

3. Khogeer, A. A., AboMansour, I. S., & Mohammed, D. A. (2022). The Role of

Genetics, Epigenetics, and the Environment in ASD: A Mini Review. Epigenomes,
6(2), 15. https://doi.org/10.3390/epigenomes6020015

4. Lyall, K., Schmidt, R. J., & Hertz-Picciotto, I. (2014). Maternal lifestyle and
environmental factors in autism. International Journal of Epidemiology, 43(2), 443-
464.

5. MacFabe, D. F. (2012). Short-chain fatty acid fermentation products of the gut

microbiome: implications in autism spectrum disorders. Microbial Ecology in Health
and Disease, 23(1), 19260.

6. Shultz, S. R., MacFabe, D. F., Martin, S., Jackson, J., Taylor, R., & Boon, F. (2008).
Intracerebroventricular injection of propionic acid, an enteric bacterial metabolic end-
product, impairs social behavior in the rat: implications for an animal model of
autism. Neuropharmacology, 54(6), 901-911.

7. World Health Organization. (2023). Traditional medicine has a long history of

contributing to conventional medicine and continues to hold promise. Retrieved from
https://www.who.int/news-room/feature-stories/detail/traditional-medicine-has-a-
long-history-of-contributing-to-conventional-medicine-and-continues-to-hold-promise

8. Kongkeaw, C., Dilokthornsakul, P., Thanarangsarit, P., Limpeanchob, N., & Norman
Scholfield, C. (2014). Meta-analysis of randomized controlled trials on cognitive
effects of Bacopa monnieri extract. Journal of Ethnopharmacology, 151(1), 528-535.

9. Singh, H. K., & Dhawan, B. N. (1997). Neuropsychopharmacological effects of the

Ayurvedic nootropic Bacopa monniera Linn. (Brahmi). Indian Journal of
Pharmacology, 29(5), S359-S365.
10. Russo, A., & Borrelli, F. (2005). Bacopa monniera, a reputed nootropic plant: A
review. Phytomedicine, 12(4), 305-317.

11. Calabrese, C., Gregory, W. L., Leo, M., Kraemer, D., Bone, K., & Oken, B. (2008).
Effects of a standardized Bacopa monnieri extract on cognitive performance, anxiety,
and depression in the elderly: A randomized, double-blind, placebo-controlled trial.
Journal of Alternative and Complementary Medicine, 14(6), 707-713.

12. Fuster, J. M. (2001). The prefrontal cortex—An update. Neuron, 30(2), 319-333.
https://doi.org/10.1016/S0896-6273(01)00285-9

13. Mohapatra, A. N., & Wagner, S. (2023). The role of the prefrontal cortex in social
interactions of animal models and the implications for autism spectrum disorder.
Frontiers in Psychiatry, 14, 1205199. https://doi.org/10.3389/fpsyt.2023.1205199