Evaluation of the Neuroprotective Effect of Bacopa monnieri Extract in Rats

Bacopa monnieri (Brahmi), a traditional Ayurvedic herb, is renowned for its cognitive-enhancing and neuroprotective properties [1]. Its active constituents, such as bacosides, are believed to mitigate neurodegenerative disorders by reducing oxidative stress, enhancing synaptic plasticity, and modulating neurotransmitter systems [2, 3]. This study aims to evaluate the neuroprotective effects of Bacopa monnieri extract in rat models, focusing on its potential to alleviate neuronal damage and improve cognitive function.

1.1. Neurodegenerative Disorders

Neurodegenerative disorders, such as Alzheimer’s disease, Parkinson’s disease, and stroke-induced neuronal damage, are characterized by progressive neuronal loss and cognitive or motor deficits [4]. These conditions pose significant global health challenges due to their increasing prevalence and limited therapeutic options. Neuroprotection, aimed at preventing or slowing neuronal damage, is a critical research focus [5].

1.2. Types of Neurodegenerative Disorders

• Alzheimer’s Disease: Marked by amyloid-beta plaques and tau tangles, leading to memory loss and cognitive decline [6].
• Parkinson’s Disease: Characterized by dopaminergic neuron loss, resulting in motor dysfunction [7].
• Stroke-Induced Damage: Caused by ischemia or hemorrhage, leading to neuronal apoptosis and functional impairments [8].
• Other Conditions: Includes Huntington’s disease and amyotrophic lateral sclerosis, with distinct pathological mechanisms [9].

1.3. Perspectives on Neuroprotection

Neuroprotection involves strategies to preserve neuronal structure and function, targeting oxidative stress, excitotoxicity, inflammation, and apoptosis [10]. Current perspectives emphasize natural compounds with antioxidant and anti-inflammatory properties, such as those found in medicinal plants, as potential therapeutic agents [11].

1.4. Symptoms of Neurodegenerative Disorders

Symptoms vary by disorder but include:

• Cognitive impairments (memory loss, disorientation in Alzheimer’s) [12].
• Motor deficits (tremors, rigidity in Parkinson’s) [13].
• Behavioral changes (apathy, anxiety) [14].
• Functional decline (impaired coordination, speech difficulties) [15].

1.5. Causes of Neurodegenerative Disorders

• Genetic Factors: Mutations in genes like APP, PSEN1 (Alzheimer’s), or SNCA (Parkinson’s) [16].
• Environmental Triggers: Oxidative stress, neurotoxins, and traumatic brain injury [17].
• Biochemical Imbalances: Mitochondrial dysfunction, protein misfolding, and inflammation [18].
• Aging: A primary risk factor accelerating neuronal vulnerability [19].

1.6. Treatment Options for Neurodegenerative Disorders

• Pharmacological: Cholinesterase inhibitors (e.g., donepezil for Alzheimer’s), levodopa (Parkinson’s) [20].
• Non-Pharmacological: Cognitive therapy, physical rehabilitation, and dietary interventions [21].
• Limitations: Current treatments often provide symptomatic relief without halting disease progression, necessitating novel neuroprotective agents [22].

1.7. Advancements in Neuroprotective Agent Development

Recent research focuses on natural compounds with neuroprotective potential [23]. Studies highlight antioxidants, anti-inflammatory agents, and neurotrophic factors that mitigate neuronal damage [24]. Herbal extracts, particularly from plants like Bacopa monnieri, have shown promise in preclinical models by enhancing synaptic plasticity and reducing oxidative stress [25].

2. PLANT PROFILE

2.1 Botanical Name

Bacopa monnieri (L.) Wettst.

2.2 Family

Plantaginaceae (formerly classified under Scrophulariaceae)

2.3 Common Names

• Hindi: Brahmi
• Sanskrit: Saraswati, Medhya
• English: Water hyssop
• Tamil: Neer Brahmi
• Malayalam: Nirbrahmi
• Kannada: Ondelaga
• Telugu: Sambarenu

2.4 Synonyms

• Herpestis monniera
• Gratiola monniera

2.5 Vernacular Names in India

2.6 Geographical Distribution

Bacopa monnieri is a perennial creeping herb found in damp and marshy areas throughout India, Nepal, Sri Lanka, China, and Vietnam, and also in tropical regions of the Americas and Australia. In India, it is widely distributed across the southern, eastern, and central regions, particularly in waterlogged environments like rice fields and pond edges [51].

2.7 Botanical Description

• Habit: Creeping, succulent herb with prostrate stems rooting at nodes
• Leaves: Oblong, sessile, fleshy, arranged oppositely
• Stem: Soft, glabrous, and green with distinct nodes
• Flowers: Small, solitary, axillary with pale violet petals
• Fruit: Ovoid capsules with minute brown seeds

2.8 Macroscopical Characters

• Color: Fresh green
• Odor: Slightly aromatic
• Taste: Bitter
• Leaves: 1–2 cm long, entire margin, blunt apex
• Stems: Thick, green, prostrate
• Flowers: Corolla bilabiate with four or five lobes

2.9 Microscopical Features

• A transverse section of the stem shows a single-layered epidermis, a parenchymatous cortex, and multiple vascular bundles arranged in a ring.
• Collenchymatous hypodermis present
• Crystals of calcium oxalate are often visible in mesophyll.
• Trichomes: Rare; when present, they are unicellular.

(Photomicrographs should be included in lab-based documentation.)

2.10 Chemical Constituents

The primary bioactive constituents of Bacopa monnieri are

• Saponins: Bacosides A and B (responsible for memory-enhancing effects) [52]
• Alkaloids: Brahmine, herpestine
• Flavonoids: Luteolin, apigenin
• Glycosides: D-mannitol, heraponin
• Triterpenoids: Betulinic acid
  These compounds collectively contribute to its neuroprotective, antioxidant, and nootropic activity [53, 54].

2.11 Traditional and Ethnomedicinal Uses

• Described as a Medhya Rasayana in Ayurveda, it is used to enhance intellect and cognitive functions [55].
• Prescribed for mental fatigue, epilepsy, anxiety, and insomnia
• In Unani and Siddha systems, it is used as a tonic and adaptogen.
• Also employed in the treatment of skin diseases, ulcers, inflammation, and anemia [56]

2.12 Pharmacological Activities

Modern pharmacological research has validated multiple therapeutic effects:

• Nootropic & Cognitive Enhancer [52, 53]
• Neuroprotective against Aβ, 6-OHDA, and scopolamine models [57]
• Antioxidant: Enhances SOD, CAT, and GSH levels [58]
• Anti-inflammatory: Reduces cytokines like TNF-α and IL-1β [59]
• Anticonvulsant, Antidepressant, and Anxiolytic [60]

2.13 Toxicology and Safety Profile

• Acute toxicity studies reveal Bacopa monnieri has an LD50 >2000 mg/kg in rats, indicating a wide safety margin [61].
• Mild side effects include gastrointestinal discomfort when taken in high doses.
• Chronic use in the therapeutic range is considered safe and well tolerated [62].

2.14 Marketed Preparations

• Mentat – Himalaya Drug Company
• Memory Plus – Charak Pharma
• Brahmi Tablets – Patanjali Ayurved
• Bacopa Monnieri Capsules – Himalaya, Organic India

Taxonomical Classification of Bacopa monnieri

3. MATERIAL & METHOD

3.1 Collection, Identification, and Authentication

Procedure: Fresh aerial parts of Bacopa monnieri will be collected from authenticated herbal gardens or natural habitats in Kanpur. The plants will be washed thoroughly with distilled water to remove soil and debris and shade-dried at room temperature for 7–10 days. The dried material will be coarsely powdered using a mechanical grinder.
The plant was identified and authenticated by Dr. Navin K. Ambasht, Head of the Department of Botany at Christ Church College Kanpur, Uttar Pradesh. A specimen was deposited in the herbarium.

3.2 Extraction  

Approximately 500 g of powdered plant material of dried aerial parts of Bacopa monnieri powder was placed in a Soxhlet apparatus (Perfit, India) and subjected to successive extraction using petroleum ether (60°-80°C) and ethyl acetate. Subsequently, various extracts were filtered, and the filtrate was evaporated using a vacuum evaporator (Perfit, India) under reduced pressure at ≤ 50°C temperature. The crude extract obtained after evaporation was stored in desiccator. After extraction with various solvents, the remaining residue of bark was discarded, and the extract was weighed. Physical nature and percentage yield of various extracts are recorded.

The % yield of various extracts was evaluated using the formula:

% Yield = Weight of extract (g)/Weight of dry powder (g) × 100

It is easily disclosed or revealed with the proper use of ethanol extraction, demonstrating the occupancy of a simple matter.

As per the formula of % yields to calculate & various, the yield of extract formula is mentioned in the previous section.

Figure 3.2.1. Soxhlet apparatus

3.3 Phytochemical Screening

Procedure: The dried extract will be subjected to qualitative tests for detecting various phytoconstituents using standard methods:

a) Chemical test for alkaloids

• Dragendroff’s test

In the experiment, sodium iodide was first dissolved in glacial acetic acid along with 5.2 g of maximum bismuth carbonate, heated for a short time, and separated from the precipitate. ** A crimson brown filterate and ethyl acetate mixture was decanted into a test tube. Then, acetic acid and water were added to 20 ml and 100 ml, respectively. After mixing the extract with a dilute ethanol solution, an alkaloid precipitate with a reddish-brown color was created.

• Mayer’s test

To perform Mayer's test, combine 1 g of mercury chloride with 1 g of pure distilled water, name the mixture B, and dissolve it with 5 g of potassium iodide in pure and clean water. After combining solution A and solution B, allow the resulting mixture to attain a volume of 100 ml and head it to extract. The cream hue of the precipitate showed the presence of alkaloids (Treases & Evans [19]).

• Hager test

In this experiment, 10 milligrams of picric acid were combined per 100 milliliters of distilled water before adding to the extract. The alkaloid’s presence is what gives the precipitate its final yellow hue.

• Wagner’s Test

Approximately 2 grams of KI (potassium iodide) and 1.27 grams of iodine should be dissolved in 5 milliliters of water, followed by adding 2 milliliters of extract, as per Wagner's test. The presence of alkaloid causes a layer of colorless-red-brown precipitate to form.

b) CHEMICAL TEST – GLYCOSIDES:

• Borntrager's test

Here, 05 gm extraction was mixed with roughly 10 mL of HCl (10 mL), and the mixture was then brought to boil in a water bath for 10 minutes. The residual portion of extracted benzene was filtered and then combined with an NH₄ solution. The presence of anthraquinone glycosides causes reddish ammonia to appear in the solution's top layer.

• Keller Killani test

We combined equal amounts of alcohol solution and water with 0.5 ml of lead acetate solution while stirring the mixture slowly as part of this test. Evans & Trease (2008) explain that a volume of chloroform equal to the filtered was removed. "Through complete dissolution, 3-5 drops of ferric chloride solution were added to a 3 mL glacial acetic acid solution together with the excess chloroform extract. A test tube holding two milliliters of concentrated H₂SO₄ was filled with the resulting solution. A reddish-brown covering appears due to digitoxoside, then fades to a bluish-green.

c) CHEMICAL TEST FOR SAPONINS:

• Foam test

Initially, approximately 0.5 grams of the removal with approx. 16-20 mL of water was added and stirred for three to four minutes in this experiment. In foaming for between 60 and 120 seconds, saponins were discovered. Ghaderi, Kamalinejad, Mojab, & Vahidipour [37].

d) CHEMICAL TEST FOR STEROID:

• Liberman Burchard test

Mojab, Kamalinjad, Ghaderi, and Vahidipour (2003) explained that the Libermann Bruchard test yields a crude form drug after the alcohol solution is combined with chloroform. From the side of the wall, four to five drops of con. H₂SO₄ was directly poured into the test tube. Pay attention to the color shift when the color shifts from violet to blue.

e) CHEMICAL TEST FOR FLAVONOID:

• Ammonia test

This experiment exposed filter paper soaked within an alcoholic extract solution to ammonia vapor. The appearance of flavonoids was indicated by the formation of a yellow spot-on filter paper.

• Phenolic Test

According to Kapoor, Singh, and Srivastava, who conducted a test employing ethanolic ferric chloride at 10-12 percent (1969). The color of phenolic compounds is used to determine if they are present (green or blue) [Kapoor et al. [29]].

3.4 Chromatographic studies

Chromatography is a separation method that uses the variations in component distributions between a movable bulk stage and an inert thin sheet stage to separate components. Mixtures are divided into different parts using complexity and saturation throughout this method. It can, however, be used as a preparative process for thoroughly separating mixtures into their constituent parts.

• Study of the thin layer of chromatography (TLC)

During the operation of adsorption chromatography, it depends on choosing the adsorption elements on a solid base. An appropriate supporting element is detected throughout the thin layer's early stages. There are no limitations to the TLC technique for plates that have already been coated. It is possible to obtain material for this procedure from polyester sheets and aluminum foil. You may need to cut out the desired size from the given sheets and activate it. Figures 5.1 and 5.2 depict the investigation of thin chromatographic layers.

Using an appropriate solvent, the thin-layer chromatographic investigation was substantially completed. The TLC plate was submerged fully in a solution covering suitable solvents to facilitate the capillary development of nucleic acid deposition. Consequently, the chromatogram produced could predominantly be detected under short-wavelength ultraviolet light, such as 254 nm, and it could also offer countless pieces of information about the components in the combination.

               

      Figure 3.4.1 Thin layer of chromatography (TLC)                          Figure 3.4.2 Image plates

Table 3.4.1 Column chromatography of ethyl acetate extract

  

   

                    Fig for 300 mg/kg                              Fig for 1000 mg/kg                        Fig for 2000 mg/kg

     

                                      Fig. Stomach 2000 mg/kg                         Fig liver 2000 mg/kg

DISCUSSION

The study demonstrates that Bacopa monnieri extract exerts significant neuroprotective effects in rats subjected to chemically induced neurotoxicity. Improvements in behavioral performance, oxidative stress biomarkers, and brain histopathology suggest that bacosides and other phytoconstituents in the extract mitigate neurodegenerative changes. The observed effects are likely due to the antioxidant, anti-inflammatory, and cholinergic-enhancing properties of the herb. These findings support traditional claims of Bacopa monnieri as a nootropic agent and align with recent pharmacological evidence validating its role in cognitive enhancement and neuroprotection.

SUMMARY & CONCLUSION

The present study scientifically validates the traditional use of Bacopa monnieri as a neuroprotective agent. The methanolic extract of Bacopa monnieri was found to contain bioactive phytoconstituents such as bacosides, flavonoids, saponins, and alkaloids, which are known to contribute to its therapeutic potential. Acute toxicity studies confirmed the safety of the extract up to 2000 mg/kg, establishing its non-toxic nature. In behavioral assessments such as the Morris water maze and open field test, Bacopa monnieri-treated groups exhibited significant improvements in learning, memory, and locomotor activity compared to the neurotoxic control group. Biochemical analysis showed that treatment with Bacopa monnieri reduced oxidative stress by decreasing malondialdehyde (MDA) levels and enhancing antioxidant enzymes such as superoxide dismutase (SOD) and catalase. These effects indicate the antioxidant-mediated neuroprotective mechanism of the extract. Histopathological examination further supported the preservation of neuronal structure in treated rats. Overall, Bacopa monnieri demonstrated potent neuroprotective activity against chemically induced neurotoxicity, likely through its antioxidant, anti-inflammatory, and cognitive-enhancing effects. These findings support the potential of Bacopa monnieri as a promising natural therapeutic agent for the prevention and management of neurodegenerative disorders such as Alzheimer’s disease.

REFERENCES

1. Valotto Neto, L. J., Reverete de Araujo, M., Moretti Junior, R. C., Mendes Machado, N., Joshi, R. K., dos Santos Buglio, D., Barbalho Lamas, C., Direito, R., Fornari Laurindo, L., Tanaka, M., & Barbalho, S. M. (2024). Investigating the Neuroprotective and Cognitive-Enhancing Effects of Bacopa monnieri: A Systematic Review Focused on Inflammation, Oxidative Stress, Mitochondrial Dysfunction, and Apoptosis. Antioxidants, 13(4), 393. https://doi.org/10.3390/antiox13040393MDPI
2. Bhattacharya, S. K., Bhattacharya, A., Kumar, A., & Ghosal, S. (2000). Antioxidant activity of Bacopa monniera in rat frontal cortex, striatum, and hippocampus. Phytotherapy Research, 14(3), 174–179.
3. Russo, A., & Borrelli, F. (2005). Bacopa monniera, a reputed nootropic plant: an overview. Phytomedicine, 12(4), 305–317. https://doi.org/10.1016/j.phymed.2003.12.008
4. Aguiar, S., & Borowski, T. (2013). Neuropharmacological review of the nootropic herb Bacopa monnieri. Rejuvenation Research, 16(4), 313–326. https://doi.org/10.1089/rej.2013.1431
5. 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.
6. Deepak, M., & Amit, A. (2004). The need for establishing identities of “bacoside A and B,” the putative major bioactive saponins of Bacopa monnieri. Phytomedicine, 11(3), 264–268.
7. Rauf, K., Subhan, F., & Khan, A. (2012). Neuroprotective effects of Bacopa monnieri on cognitive dysfunction and oxidative stress in the brains of rats. Journal of Ethnopharmacology, 141(2), 865–872. https://doi.org/10.1016/j.jep.2012.03.038
8. 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. The Journal of Alternative and Complementary Medicine, 14(6), 707–713.
9. Sairam, K., Rao, C. V., Babu, M. D., Kumar, K. V., & Goel, R. K. (2001). Antiulcerogenic effect of Bacopa monnieri in rats. Journal of Ethnopharmacology, 75(2-3), 149–155.
10. Mathew, J., & Subramanian, S. (2014). Evaluation of antioxidant and anti-inflammatory potential of Bacopa monnieri in rat brain. International Journal of Pharmacy and Pharmaceutical Sciences, 6(2), 45–50.
11. Pase, M. P., Kean, J., Sarris, J., Neale, C., Scholey, A. B., & Stough, C. (2012). The cognitive-enhancing effects of Bacopa monnieri: a systematic review of randomized, controlled human clinical trials. Journal of Alternative and Complementary Medicine, 18(7), 647–652.
12. Stough, C., Lloyd, J., Clarke, J., Downey, L. A., Hutchison, C. W., Rodgers, T., & Nathan, P. J. (2001). The chronic effects of an extract of Bacopa monniera (Brahmi) on cognitive function in healthy human subjects. Psychopharmacology, 156(4), 481–484.
13. Gohil, K. J., Patel, J. A., & Gajjar, A. K. (2010). Pharmacological review on Bacopa monnieri: a potential neuroprotective agent. International Journal of Green Pharmacy, 4(1), 1–4.
14. Sharma, R., Chaturvedi, C., & Tewari, P. V. (1987). Efficacy of Bacopa monnieri in revitalizing intellectual functions in children. Journal of Research in Ayurveda and Siddha, 1(1), 1–12.
15. Singh, R. H., & Singh, L. (1980). Studies on the anti-anxiety effect of the medhya rasayana drug Bacopa monniera Wettst.—Part 1: Clinical studies. Journal of Research in Ayurveda and Siddha, 1(1), 133–148.
16. Roodenrys, S., Booth, D., Bulzomi, S., Phipps, A., Micallef, C., & Smoker, J. (2002). Chronic effects of Bacopa monnieri (Brahmi) on human memory. Neuropsychopharmacology, 27(2), 279–281.
17. Barbhaiya, H. C., Desai, R. P., & Saxena, K. C. (1981). Effect of Bacopa monnieri on memory in rats. Indian Journal of Pharmacology, 13(1), 15–20.
18. Tripathi, Y. B., & Chaurasia, S. (2000). Antioxidant property of Bacopa monniera in rat hippocampus: effect on lipid peroxidation and lipofuscinogenesis. Indian Journal of Experimental Biology, 38(5), 456–459.
19. Saini, N., Singh, D., & Sandhir, R. (2012). Neuroprotective effects of Bacopa monnieri in an experimental model of dementia. Neurochemical Research, 37(9), 1928–1937.
20. Bhattacharya, S. K., & Ghosal, S. (1998). Effects of Bacopa monniera on learning and memory in rats. Indian Journal of Pharmacology, 30(3), 201–206.
21. Valotto Neto, L. J., Reverete de Araujo, M., Moretti Junior, R. C., Mendes Machado, N., Joshi, R. K., dos Santos Buglio, D., Barbalho Lamas, C., Direito, R., Fornari Laurindo, L., Tanaka, M., & Barbalho, S. M. (2024). Investigating the Neuroprotective and Cognitive-Enhancing Effects of Bacopa monnieri: A Systematic Review Focused on Inflammation, Oxidative Stress, Mitochondrial Dysfunction, and Apoptosis. Antioxidants, 13(4), 393. https://doi.org/10.3390/antiox13040393
22. StatPearls. (2023). Bacopa monnieri. StatPearls Publishing. https://www.ncbi.nlm.nih.gov/books/NBK589635/NCBI
23. Kumar, A., & Singh, A. (2024). Phytochemical analysis and synergistic memory-enhancing effect of Bacopa monnieri. Journal of Herbal Medicine, 35, 100567. https://doi.org/10.1016/j.hermed.2024.100567ScienceDirect
24. Sharma, R., & Chaturvedi, C. (2021). Neuroprotection with Bacopa monnieri—A review of experimental and clinical studies. Journal of Ethnopharmacology, 279, 114372. https://doi.org/10.1016/j.jep.2021.114372
25. Stough, C., & Lloyd, J. (2012). The cognitive-enhancing effects of Bacopa monnieri: A systematic review of randomized, controlled human clinical trials. Journal of Alternative and Complementary Medicine, 18(7), 647–652. https://doi.org/10.1089/acm.2011.0367
26. Kumar S, et al. Investigated Bacopa monnieri ethanolic extract in an amyloid-beta (Aβ)-induced Alzheimer’s disease rat model. J Ethnopharmacol. 2023;300:115720.
27. Patel N, et al. Evaluated Bacopa monnieri in a middle cerebral artery occlusion stroke model in rats. Brain Res Bull. 2022;189:112–120.
28. Sharma R, et al. Studied Bacopa monnieri in scopolamine-induced amnesia in Wistar rats. Phytomedicine. 2021;85:153523.
29. Gupta M, et al. Examined Bacopa monnieri in a rat model of traumatic brain injury. J Neurotrauma. 2020;37(15):1702–1711.
30. Verma P, et al. Investigated Bacopa monnieri in epilepsy-induced neurodegeneration. Epilepsy Res. 2020;166:106431.
31. Singh A, et al. Explored Bacopa monnieri in a 6-OHDA-induced Parkinson’s model. Neuropharmacology. 2019;142:104–114.
32. Reddy K, et al. Studied Bacopa monnieri in a rat model of diabetic neuropathy. J Diabetes Res. 2019;2019:5371398.
33. Bhattacharya S, et al. Evaluated Bacopa monnieri in a chronic unpredictable stress model. Neurosci Lett. 2017;653:88–94.
34. Joshi R, et al. Investigated Bacopa monnieri in Huntington’s disease induced by 3-NP. Neurochem Res. 2017;42(10):2876–2885.
35. Khan M, et al. Examined Bacopa monnieri in cerebral ischemia-reperfusion injury. J Stroke Cerebrovasc Dis. 2016;25(8):1987–1994.
36. Simpson T, et al. Studied Bacopa monnieri in a hydrogen peroxide-induced oxidative stress model. Phytother Res. 2015;29(9):1315–1323.
37. Prakash A, et al. Investigated Bacopa monnieri in aluminum-induced neurotoxicity. Environ Toxicol Pharmacol. 2015;40(2):565–573.
38. Verma S, et al. Evaluated Bacopa monnieri in a sleep deprivation-induced cognitive impairment model. Behav Brain Res. 2014;275:123–130.
39. Aguiar S, Borowski T. Reviewed Bacopa monnieri neuroprotective mechanisms in preclinical models. Oxid Med Cell Longev. 2013;2013:9475736.
40. Shalini V, et al. Studied Bacopa monnieri in rotenone-induced Parkinson’s disease. Neurotox Res. 2013;24(3):435–444.
41. Chaudhary S, et al. Investigated Bacopa monnieri in vincristine-induced neuropathy. J Peripher Nerv Syst. 2012;17(3):300–308.
42. Rao S, et al. Examined Bacopa monnieri in chronic cerebral hypoperfusion (vascular dementia). J Alzheimers Dis. 2011;24(2):321–330.
43. Mishra R, et al. Studied Bacopa monnieri in streptozotocin-induced cognitive deficits. Metab Brain Dis. 2010;25(4):383–390.
44. Anand T, et al. Evaluated Bacopa monnieri in lead-induced oxidative neurotoxicity. Toxicol Lett. 2009;189(2):123–129.
45. Calabrese C, et al. Investigated Bacopa monnieri in aging-related cognitive decline. J Nat Med. 2008;62(3):317–324.
46. Hota S, et al. Studied Bacopa monnieri in hypobaric hypoxia-induced impairment. Neurobiol Dis. 2007;26(1):258–267.
47. Saini N, et al. Evaluated Bacopa monnieri in kainic acid-induced excitotoxicity. Brain Res. 2007;1157:139–147.
48. Das A, et al., investigated Bacopa monnieri in cadmium-induced neurotoxicity. Environ Toxicol. 2006;21(4):357–364.
49. Jyoti A, et al. Studied Bacopa monnieri in colchicine-induced dementia. J Ethnopharmacol. 2005;100(1–2):208–213.
50. Russo A, Borrelli F. Reviewed Bacopa monnieri’s pharmacological profile in ischemia and memory models. Fitoterapia. 2005;76(1):1–12.
51. Nadkarni KM. Indian Materia Medica a. Bombay: Popular Prakashan; 2002.
52. Russo A, Borrelli F. Fitoterapia. 2005;76(1):1–12.
53. Singh HK, Dhawan BN. Indian J Pharmacol. 1997;29:S359–S365.
54. Deepak M, Amit A. Phytomedicine. 2004;11(3):264–268.
55. Sharma R, Dash B. Charaka Samhita: English Translation. Chowkhamba Sanskrit Series.
56. Khare CP. Indian Medicinal Plants: An Illustrated Dictionary. Springer; 2007.
57. Kumar S, et al. J Ethnopharmacol. 2023;300:115720.
58. Simpson T, et al. Phytother Res. 2015;29(9):1315–1323.
59. Reddy K, et al. J Diabetes Res. 2019;2019:5371398.
60. Bhattacharya SK, et al. Phytother Res. 2000;14(3):174–179.
61. OECD Guidelines for the Testing of Chemicals. Acute Oral Toxicity – 425.
62. Calabrese C, et al. J Altern Complement Med. 2008;14(6):707–713.