This result was attributed to the Aegle marmelos, a commonly known antidiarrheal in India, although the two botanicals were not given separately, so individual effects cannot be confirmed. Ayurvedic therapy was superior to placebo in all parameters examined, but no benefit could be linked specifically to the Bacopa portion of the Ayurvedic preparation.³⁴ Animal and in vitro studies suggest Bacopa may have a protective and curative effect for gastric ulcers. In rats a Bacopa extract standardized for bacoside A was evaluated for its prophylactic and healing effects in five models of gastric ulcers. At a dose of 20 mg/kg for 10 days, Bacopa extract significantly healed penetrating ulcers induced by acetic acid, significantly strengthened the mucosal barrier, and decreased mucosal exfoliation. The extract also alleviated stress-induced ulcers as observed by significant reduction in lipid peroxidation in rat gastric mucosa. Bacopa’s antioxidant properties and its ability to balance SOD and catalase levels may account for this effect.³⁵ A recent in vitro study also demonstrated Bacopa extract’s specific anti-microbial activity against Helicobacter pylori, a bacteria associated with chronic gastric ulcers. When the extract was incubated with human colonic muscosal cells and H. pylori it resulted in accumulation of prostaglandin E and prostacycline,prostaglandins known to be protective for gastric mucosa.³⁶In another placebo-controlled study, 169 patients with irritable bowel syndrome were given a Bacopa preparation. They experienced considerable improvement: 65%, compared to 33% in the control group.

CARDIOVASCULAR EFFECTS:

Use of Bacopa as a “cardiotonic” is frequently mentioned in Ayurvedic medicine texts, but no clinical studies have been conducted. In vitro research using rabbit aorta and pulmonary artery has demonstrated Bacopa extract exerts a vasodilatory effect on calcium chloride-induced contraction in both tissues. It is believed to exert this effect via interference with calcium channel flux in tissue cells.³⁷In addition to these findings, just four weeks of Bacopa therapy resulted in a decrease in average systolic blood pressure, from 117 mmHg to 112 mmHg, and a significant improvement in respiratory function: breath-holding time increased from 35 seconds to 48 seconds (a 37% increase). Patients also showed a reduction in adrenocortical function, indicating a less stressed state. As well, notable relief was found from other symptoms of anxiety: insomnia, headache, irritability, lack of concentration, anorexia, dyspepsia, tremors, palpitation, and nervousness. Unlike other antianxiety compounds, Bacopa was thought by the researchers to increase nonspecific overall resistance to stress (the so-called "adaptogenic" effect), allowing the system to adapt effortlessly to a work overload, for example.

HYPERTHYROIDISM:

A study in mice demonstrated high doses (200 mg/kg) of Bacopa extract increased the thyroid hormone, T4, by 41 percent when given orally. T3 was not stimulated, suggesting the extract may directly stimulate synthesis and/or release of T4 at the glandular level, while not affecting conversion of T4 to T3. While this study indicates Bacopa extract does have a stimulatory effect on thyroid function, the doses were very high and the typical 200-400 mg daily dose in humans may not have the same effect.³⁸

PROTECTION FROM DRUG TOXICITY:

In vitro and animal studies have demonstrated Bacopa extracts may have a protective effect against certain drugs and their negative side effects. An in vitro study using guinea pig ileum isolates examined the effect of Bacopa extract on drug-induced morphine withdrawal. Addition of 1,000 g/mL Bacopa extract to the tissue isolates prior to injection of morphine significantly reduced the naloxone-induced withdrawal effects,³⁹an effect that may be attributed to the anticholinergic and calcium antagonistic activity reported by other researchers.⁴⁰ A second study examined the effects of an alcohol extract of Bacopa on morphine-induced hepatotoxicity in rats, as measured by lipid peroxide accumulation and antioxidant enzyme levels. Administration of Bacopa extract with morphine significantly decreased lipid peroxidation and increased levels of antioxidant enzymes and glutathione in rat hepatic tissue, when compared to morphine alone. These results suggest a protective effect for Bacopa on the hepatic antioxidant status in morphine-treated rats.⁴¹ Some of the same researchers reported a similar effect for brain mitochondrial enzyme activity of morphinetreated rats.⁴² It has also been reported that antiepileptic drugs, such as phenytoin, can result in cognitive impairment.⁴³ In mice, Bacopa administration with phenytoin significantly reversed phenytoin-induced cognitive impairment, as noted by improved acquisition and retention of memory. These results suggest a potential corrective effect of Bacopa extracts in phenytoin-induced cognitive deficit.⁴⁴This study and others indicate that the toxicity of Bacopa is very low. A person weighing just 60 kg (132 lbs) would have to take more than 1 kg (2.2 lbs) of the extract daily to produce a dangerous reaction. The actual suggested dose of Bacopa is 60 mg three times a day, or 180 mg/day, so there is clearly a very high safety margin.

CANCER:

In vitro research demonstrated Bacopa saponin fractions have cytotoxic activity for sarcoma-180 cells. It is thought this might be due to Bacopa’s inhibition of DNA replication in the cancerous cell line.⁴⁵ Research in humans may be indicated.

ANTIOXIDANT ACTIVITY:

Ethanolic extract of B. monnieri is found to contain phenolic compound in significant amount, which attributes to its rationality of possessing antioxidant activity. For measurements of the reductive ability, we investigated Fe+3 to Fe+2 transformations in the presence of ethanolic extract. The reducing capacity of the extract may serve as a significant indicator of its potential antioxidant activity ⁴⁶. Nitric oxide (NO) exhibits numerous physiological properties and it is also implicated in several pathological states ⁴⁷. The decrease in the nitrite production could also be due to interaction of the extract with other nitrogen oxides ⁴⁸. The in vitro superoxide radical scavenging activity is measured by riboflavin/ light/ NBT (Nitroblue tetrazoline) system reduction. The method is based on generation of superoxide radicals by auto oxidation of riboflavin in the presence of light. The superoxide radical reduces NBT to a blue colored formazone that can be measured at 560 nm⁵⁰. The capacity of the ethanolic extract of B. monnieri to inhibit the colour to 50% is measured in terms of IC50. Superoxide radical is known to be very harmful to the cellular components as a precursor of more ROS⁵⁰. The extract has been found to have significant superoxide radical scavenging activity, which ultimately adds to its antioxidant potential. One study examined how it might help prevent induced lipid peroxidation.⁵¹ Its protective capacity was found to compare favorably to the effects of the chelating agent EDTA and to those of the natural antioxidant vitamin E. The researchers could not discern whether Bacopa's principal mechanism of action was related to its metal-chelating or antioxidant properties. Bacopa is a potent antioxidant. It has been suggested that its role in memory enhancement may be due in part to its ability to increase brain circulation by inhibiting oxidative damage in the brain. It also enhances serotonin levels in the brain.

HEPATOPROTECTIVE ACTIVITY:

Paracetomol produces hepatic necrosis when ingested in very large doses. The hepatoprotective activity of the ethanolic extract was monitored by estimating serum transaminases, serum alkaline phosphatase and bilirubin, which give a good idea about the functional state of the liver ⁵². Necrosis or membrane damage releases the enzyme into circulation and therefore, it can be measured in serum. High levels of SGOT indicate liver damage, such as that due to viral hepatitis as well as cardiac infarction and muscle injury. SGPT catalyses the conversion of alanine to pyruvate and glutamate, and is released in a similar manner. Therefore, SGPT is more specific to the liver, and is thus a better parameter for detecting liver injury ⁵³. Our results demonstrate that the ethanolic extract of B. monnieri caused signifi-cant inhibition of SGOT and SGPT levels. Serum ALP and bilirubin levels on the other hand, are related to the function of hepatic cell. Increase in serum level of ALP is due to increased synthesis, in presence of increasing biliary pressure ⁵⁴. Our results also demonstrate that the ethanolic extract of B. monnieri caused significant inhibition of serum ALP and bilirubin levels. Effective control of alkaline phosphatase activity and bilirubin level points towards an early improvement in secretory mechanism of hepatic cells. The paracetamol treated rats showed a significant loss in liver weight. But the extract admini-stration significantly prevented this paracetamol-induced weight loss of liver in rats, which is comparable with that of standard drug administration.

Most hepatotoxic chemicals including paracetamol and alcohol damage liver by inducing, directly or indi-rectly, lipid peroxidation ⁵⁵. So, the studies on in vitro and in vivo lipid peroxidation of rats are performed. In vitro lipid peroxidation in liver homogenate can proceed in a non-enzymatic way. The process is induced by ascorbate in the presence ofFe2+/Fe3+, and it has been reported that Fe2+ and ascorbic acid stimulated lipid peroxidation in rat liver microsomes and mitochondria. In order to clarify the mode of action of ethanolic extract, in vitro lipid peroxidation experiments were carried out. According to the result obtained, the extract inhibited FeCl2-ascorbic acid-stimulated lipid peroxidation in liver homogenate . Cells have a number of mechanisms to protect themselves from the toxic effects of ROS. SOD removes superoxide (O- 2) by converting it to H2O2, which can be rapidly converted to water by CAT ⁵⁶. In addition, a large reserve of reduced glutathione is present in hepatocytes and red blood cells for detoxification of xenobiotics or free radicals. However, oxidative stress results in toxicity when the rate at which the ROS are generated exceeds the cell capacity for their removal. Lipid peroxidation is an autocatalytic process, which is a common consequence of cell death. This process may cause peroxidative tissue damage in inflammation, cancer and toxicity of xenobiotics and aging. MDA is one of the end products in the lipid peroxidation process ⁵⁷. In our in vivo study elevation in levels of end products of lipid peroxidation in liver of rats treated with paracetamol were observed. The increase in MDA levels in liver suggests enhanced lipid peroxidation leading to tissue damage. Pretreatment with EBM significantly reversed these changes. Hence it may be possible that the mechanism of hepatoprotection of extract is due to its antioxidant effect. GSH is widely distributed in cells. GSH is an intracellular reductant and plays major role in catalysis, metabolism and transport. It protects cells against free radicals, peroxides and other toxic compounds. Indeed, GSH depletion increases the sensitivity of cells to various aggressions and also has several metabolic effects, for example, a decrease in the rate of gluconeogenesis or an increase in glycogenolysis. The concept of a glutathione- SH threshold for drug detoxification was discussed by Jollow ⁵⁸. GSH is a naturally occurring substance that is abundant in many living creatures. It is widely known that a deficiency of GSH within living organisms can lead to tissue disorder and injury. For example, liver injury included by consuming alcohol or by taking drugs like acetaminophen, lung injury by smoking and muscle injury by intense physical activity ⁵⁹, all are known to be correlated with low tissue levels of GSH. From this point of view, exogenous ethanolic extract of B. monnieri supplementation might provide a mean of recover reduced GSH levels and to prevent tissue disorders and injuries. In the present study, we have demonstrated the effectiveness of the extract by using paracetamol-induced rats, which is a known model for both hepatic GSH depletion and injury. Therefore, the levels of glutathione are of critical importance in liver injury caused by paracetamol. Our results are in line with this earlier report because we found that after EBM -supplementation elevated GSH level in rats with paracetamol could be blunted to normal level. Biological systems protect themselves against the damaging effects of activated species by several means. These include free radical scavengers and chain reaction terminators, enzymes such as SOD and CAT. ⁶⁰ The SOD dismutases superoxide radicals O2 - into H2O2 plus O2, thus participating with other antioxidant enzymes, in the enzymatic defense against oxygen toxicity. In this study, SOD plays an important role in the elimination of ROS derived from the peroxidative process of xenobiotics in liver tissues. The observed increase of SOD activity suggests that the ethanolic extract of B. monnieri have an efficient protective mechanism in response to ROS. And also, these findings indicate that the extract may be associated with decreased oxidative stress and free radical- mediated tissue damage. CAT is a key component of the antioxidant defense system. Inhibition of these protective mechanisms results in enhanced sensitivity to free radical-induced cellular damage. Excessive production of free radicals may result in alterations in the biological activity of cellular macromolecules. Therefore, the reduction in the activity of these enzymes may result in a number of deleterious effects due to the accumulation of superoxide radicals and hydrogen peroxide. Administration of ethanolic extract of B. monnieri increases the activities of catalase in paracetamolinduced liver damage rats to prevent the accumulation of excessive free radicals and protects the liver from paracetamol intoxication. This ability of EBM to protect the liver from paracetamol-induced damage might be attributed to its ability to restore the activity of antioxidative enzymes. Thus, results of these studies together with those of earlier ones, suggest that EBM has an ability to protect the liver from paracetamol-induced damage through its direct antioxidative effect. It is well documented that hepatocellular enzymes (SOD, CAT) serve as biomarkers of hepatocellular injury due to alcohol and drug toxicity ⁶¹. So the studies on antioxidant enzymes (SOD, CAT) have been found to be of great importance in assessment of liver damage. In summary, we demonstrate that EBM prevents paracetamol-induced oxidative stress and hepatic injury. Since these models of hepatic damage in the rat simulate many of the features of human liver pathology, we suggest that natural antioxidants and scavenging agents in BME might be effective as plant hepatoprotectors and thus may have some obvious therapeutic implications. Therefore, it seems logical to infer that EBM, because of its antioxidant property, might be capable of protect-ing the hepatic tissue from paracetamol-induced injury and inflammatory changes. The ethanolic extract of B. monnieri is reported to be rich in saponins ⁶².

ATTENTION-DEFICIT-DISORDER:

Another interesting application comes from clinical reports of Bacopa's use for attention-deficit hyperactivity disorder (ADHD) in children. In a study conducted at BRD Medical College at Gorakhpur, 36 children in the 8-10-year age group were selected for a double-blind, randomized trial.⁶³ While 19 were given 50 mg of Bacopa twice daily, 17 others received placebo. After 12 weeks of treatment, the children were subjected to a battery of specialized tests. The data revealed a significant improvement in the areas of sentence repetition, logical memory, and pair-associative learning (matching things that go together; e.g., "test" and "grade") in all 19 ADHD children who took Bacopa. Evaluation did not occur until four weeks after withdrawal from Bacopa usage, indicating that it had a lasting effect.

REVITALIZING INTELLECTUAL FUNCTIONS IN CHILDREN:

As researcher Paris Kidd reports,⁶⁴Bacopa has been used traditionally to consecrate newborns in the hope of improving their intelligence, to "open the gate of Brahma." This is similar to the use of ephedra juice, believed to be the drink of longevity that ancient Aryans also gave to their newborns a custom mentioned in the Vedic text the Rig-Veda.⁶⁵ Even nowadays, Bacopa is given to schoolchildren for the same purpose, without apparent side effects. In a fascinating study, Sharma and colleagues gave one-half of a group of 40 healthy children (ages 6-8) Bacopa in a syrup base three times a day (a total of 1.05 g/day) over the course of four weeks, while giving the other half a placebo.⁶⁶ Using standard psychological tests to measure eye-hand coordination and perception abilities, as well as memory span and visual motor ability, what the researchers found was interesting. Those children taking Bacopa were superior in matters of speed and accuracy in solving maze problems. Overall, these improvements "vitalized" the children's efficacy and their propensity to choose exploratory behavior and to opt for novel experiences in preference to familiar ones. This function is regarded as central to curiosity, a complex function that is related to a number of other attributes, such as motivation, arousal, attention, and novelty preference. Exploratory behavior may also be related to lifespan.⁶⁷ When it takes hold in the early years (the earlier the better), the result can be integrative. Psychologically, as we grow older, exploratory behavioral development is likely to channel into and serve both libidinal and aggressive aims. Sharma and colleagues would have us believe that Bacopa alters psychology, not unlike a great teacher, or the impact of an extraordinary experience. Interesting, but not substantiated. Immediate memory and perception and the children's reaction/performance times were also found to be changed by Bacopa. Short-term memory and recall were significantly improved too. On yet another front, Bacopa vitalized the children's ability to handle increasingly difficult tasks. At the same time, vigilance and attention were enhanced - skills thought necessary to organize perceptions and improve perceptual motor functions. Developmentally, vigilance and attention skills are very important. The Bacopa studies were done with children between the ages of 6 and 8.Bacopa is a useful agent for enhancing and vitalizing the psychology of intellectual behavior. The natural agent Bacopa can enhance intelligence and intellectual functions traditionally claimed to be genetic. Continuing the work of Sharma et al., just recently, at the Nair Children's Hospital in Bombay, another placebo-controlled, double-blind study tested the efficacy of Bacopa on children.⁶⁸ For six weeks, 50 normal schoolchildren split into two groups were given either Bacopa or placebo. At the conclusion, they were evaluated for attention, concentration, and memory. Bacopa was shown to improve mean reaction time (auditory and visual) significantly.

REDUCTION OF PAIN:

Another study has found that the pain response caused by hypersensitive nerve endings is decreased after using Bacopa.²³ This response can be caused by either an intestinal or central nervous system condition. The latter is usually associated with cancer pain, meaning that Bacopa may be able to alleviate some of the pain in patients with this disease. Bacopa also produces tranquilizing effects, which add to its adaptogenic and antidepressant benefits. And there are virtually no side effects.

REFERENCE:

29. Dar A, Channa S. Relaxant effect of ethanol extract of Bacopa monniera on trachea, pulmonary artery and aorta from rabbit and guinea-pig. Phytother Res1997;11:323-325.

30. Channa S, Dar A, Yaqoob M, et al. Bronchovasodilatory activity of fractions and pure constituents isolated from Bacopa monniera. J Ethnopharmacol 2003;86:27-35.

31. Samiulla DS, Prashanth D, Amit A. Mast cell stabilizing activity of Bacopa monnieri. Fitoterapia 2001;72:284-285.

32. Dar A, Channa S. Relaxant effect of ethanol extract of Bacopa monniera on trachea, pulmonary artery and aorta from rabbit and guinea-pig. Phytother Res1997;11:323-325.

33. Dar A, Channa S. Calcium antagonistic activity of Bacopa monniera on vascular and intestinal smooth muscles of rabbit and guinea-pig. J Ethnopharmacol 1999;66:167-174.

34. Yadav SK, Jain AK, Tripathi SN, Gupta JP. Irritable bowel syndrome: therapeutic evaluation of indigenous drugs. Indian J Med Res 1989;90:496-503.

35. Sairam K, Rao CV, Babu MD, Goel RK. Prophylactic and curative effects of Bacopa monniera in gastric ulcer models. Phytomedicine 2001;8:423-430.

36. Goel RK, Sairam K, Babu MD, et al. In vitroevaluation of Bacopa monniera on anti-Helicobacter pylori activity and accumulation of prostaglandins. Phytomedicine 2003;10:523-527.

37. Dar A, Channa S. Calcium antagonistic activity of Bacopa monniera on vascular and intestinal smooth muscles of rabbit and guinea-pig. J Ethnopharmacol 1999;66:167-174.

38. Kar A, Panda S, Bharti S. Relative efficacy of three medicinal plant extracts in the alteration of thyroid hormone concentrations in male mice. J Ethnopharmacol 2002;81:281-285.

39. Sumathi T, Nayeem M, Balakrishna K, et al.Alcoholic extract of Bacopa monniera reduces the in vitro effects of morphine withdrawal in guineapig ileum. J Ethnopharmacol 2002;82:75-81.

40. Dar A, Channa S. Calcium antagonistic activity of Bacopa monniera on vascular and intestinal smooth muscles of rabbit and guinea-pig. J Ethnopharmacol 1999;66:167-174.

41. Sumathy T, Subramanian S, Govindasamy S, et al.Protective role of Bacopa monniera on morphineinduced hepatotoxicity in rats. Phytotherapy Res 2002;15:643-645.

42. Sumathy T, Govindasamy S, Balakrishna K,Veluchamy G. Protective role of Bacopa monniera on morphine-induced brain mitochondrial enzyme activity in rats. Fitoterapia 2002;73:381-385.

43. Smith DB. Cognitive effects of anti-epileptic drugs.Adv Neurol 1991;55:197-212.

44. Vohora D, Pal SN, Pillai KK. Protection from phenytoin-induced cognitive deficit by Bacopa monniera, a reputed Indian nootropic plant. J Ethnopharmacol 2000;71:383-390.

45. Elangovan V, Govindasamy S, Ramamoorthy N,Balasubramaanian K. In vitro studies on the anticancer activity of Bacopa monnieri. Fitoterapia 1995;66:211-215.

46. Heir S, Kanner J, Akiri B, Hadas SP. (1995) J. Agri. Food Chem. 43: 1813.

47. Marletta MA. Nitric oxide: Biosynthesis and biological significance. Trends Biol Sci 1989; 14: 488-492.

48. Baheti JR, Kumar V, Shah GB, Goyal RK. (2005) J. Nat.Rem,5(1): 15 (2005).

49. Seth UK, Dadkar NK, Kamat UG, Selected Topics in Experi-mental Pharmacology, 1st ed, Bombay. Kothari Book Depot..1972. p. 126.

50. Halliwell B, Gutteridge JMC. In: Free radicals, ageing and dis-ease. Free radicals in Biology and Medicine, 2nd edition. Oxford. Clarendron Press. 1985. p. 279-315.

51. Tripathi YB, Chaurasia S, Tripathi E, Upadhyay A, Dubey GP. Bacopa monniera Linn. as an antioxidant: mechanism of action. Indian J Exp Biol 1996 Jun;34(6):523-6.

52. Rao KS, Mishra SH. Anti-inflammatory and hepatoprotective activities of Sida rhombifolia Linn. Indian J Pharmacol 1997; 29: 110-116.

53. Willianson EM, Okpako DT, Evans FJ. Selection, preparation and pharmacological evaluation of plant material, England. John Wiley. 1996.

54. Moss DW, Butterworth PJ. Enzymology and Medicine, London. Pitman Medical. 1974. p. 13

55. Asha VV. Preliminary studies on hepatoprotective activities of Momordica sabangulata and Naragama alata. Indian J Pharmacol 2001; 33: 276-279.

56. Halliwell B, Gutteridge JM, Cross CE. Free radicals, antioxi-dants, and human disease: where are we now? J Lab Clin Med 1992; 119: 598-620.

57. Kurata M, Suzuki M, Agar NS. Antioxidant systems and eryth-rocyte life span in mammals. Biochem Physiol 1993; 106: 477-487.

58. Jollow DJ. Glutathione thresholds in reactive metabolite toxicity. Arch Toxicol Suppl 1980; 3: 95-110.

59. Leeuwenburgh C, Ji LL. Glutathione depletion in rested and exercised mice: biochemical consequence and adaptation. Arch BiochemBiophys 1995; 316: 941-949.

60. Proctor PH, McGinness JE. The function of melanin. Arch Der-matol 1986; 122: 507-508.

61. Chottopadhyay RR, Bondyopadhyay M. Possible mechanism of hepatoprotective activity of Azadirachta indica leaf extract against paracetamol-induced hepatic damage in rats: Part III..Indian J Pharmacol 2005; 37: 184-185.

62. Singh HK, Dhawan BN. Neuropsychopharmacological effects of the Ayurvedic nootropic Bacopa monnieri Linn. (Brahmi). In-dian J Pharmacol 1997; 29: S359- S365.

63. Mishra M. Memory Plus works, claim clinical studies. The Times of India, March 29, 1998 .

64. Kidd PM. A review of nutrients and botanicals in the integrative management of cognitive dysfunction. Altern Med Rev 1999 Jun;4(3):144-61.

65. Mahdihassan S, Mehdi FS. Soma of the Rigveda and an attempt to identify it. Am J Chin Med 1989;17(1-2):1-8.

66. Sharma R, Chaturvedi C, Tewari PV. Efficacy of Bacopa monierra in revitalizing intellectual functions in children. J Rees Edu Ind Med 1987:1-12.

67. Mayes LC. Exploring internal and external worlds. Reflections on being curious. Psychoanal Study Child 1991;46:3-36.

68. Kaur BR, Adhiraj J, Pandit PR, Ajita R, Vijay M, Shanta D, Hemangeeni D, Sudha M, Kamble G. Effect of an Ayurvedic formulation on attention, concentration and memory in normal school-going children. Indian Drugs 1998;35(4):200-3.

Received on 26.02.2010

Accepted on 30.04.2010

Research Journal of Pharmacognosy and Phytochemistry. 2(3): May-June 2010, 188-192