The search was performed through electronic databases such as PubMed, Scopus, Web of Science, and Google Scholar. Studies published between 2000 and 2026 were given priority. The Medical Subject Headings browser and keyword search were used. The key search terms included “cerebral ischemia,” “Mucuna pruriens,” “oxidative stress,” “animal model,” and “neuroprotection.”

Initially, articles were selected based on their titles and abstracts. This was subsequently followed by downloading the full texts to read the manuscripts completely.

In animal model studies of cerebral ischemia, studies evaluating any extract of M. pruriens and reporting biochemical, histopathological, and behavioral outcomes were included. Peer-reviewed scientific journal publications, including original animal model research, original clinical research, systematic review articles, and narrative reviews published in the English language, were the inclusion criteria for this study.

Editorials, nonischemic models, non-extraction studies of M. pruriens, in vitro-only studies, articles without accessible full text, conference proceedings, letters to the editor, commentaries, non-English-language publications, articles with insufficient methodological clarity, and duplicate data were excluded from the present study.

Three authors reviewed the articles included in the literature search.

Ischemic brain injury involves complex biochemical, molecular, and cellular mechanisms. The cascade of events includes necrosis, apoptosis, the autoimmune response, and neuroplasticity [27]. Age-based variation has been reported to affect neuroprotection and ischemic outcomes [28]. Increased levels of neurotransmitters, including glutamate, contribute to excitation of receptors and subsequent neuronal injury. Disturbance in calcium ion homeostasis aggravates cell injury [29]. Astrocytic and microglial activation, along with infiltration of neutrophils and lymphocytes, leads to amplification of the inflammatory response [30]. The pathophysiology of cerebral ischemia is represented in Figure 1 [3, 5, 7, 8, 11, 23].

Research using animal models of ischemia suggests that reactive gliosis results from a response to altered perfusion. Changes in blood flow lead to glial activation and subsequent gliosis [31]. However, when chronic reactive gliosis occurs, neuronal recovery and regeneration are inhibited. Chronic astrogliosis is associated with the formation of glial scarring, which creates physical and molecular barriers that impede axon regeneration and functional recovery [32]. Microglial activation also triggers astrogliosis through the release of cytokines, which can exacerbate astrocytes’ reactive state, furthering neuronal injury.

Immediately after the reduction in blood flow, oxygen and glucose levels decrease, further reducing adenosine triphosphate and energy-dependent processes [33]. Chronic cerebral ischemia occurs because of long-term reduction in cerebral blood flow caused by conditions such as hypertension and atherosclerosis, resulting in impaired cognition and dementia. Ionic imbalance occurs with the influx of sodium and calcium into cells and the involvement of the mitochondrial permeability transition pore. Nitric oxide and peroxynitrite also play important roles in the etiopathogenesis of ischemia–reperfusion injury.

Restoration of the blood supply can paradoxically aggravate injury through oxidative stress, inflammation, and disruption of the blood–brain barrier [30]. Leukocytic infiltration and activation of platelets and the complement system play instrumental roles [34]. Ischemia followed by reperfusion can affect mitochondria, leading to dysfunction. Injury to the mitochondrial complex and oxidative modifications are crucial aspects of the pathogenesis of ischemic neuronal injury [35]. The NLRP3 inflammasome and the Nrf2–Keap1 pathway are important during the management of reperfusion injury.

Autophagy impairment eventually leads to the accumulation of damaged organelles and protein misfolding and aggregation [36]. The proinflammatory (M1) and anti-inflammatory (M2) phenotypes of microglial polarization play crucial roles in neuroinflammation [30]. Release of chemokines and inflammatory cytokines further contributes to mediation of neuronal injury and disruption of the blood–brain barrier. The LC3-II, Beclin-1, and p62 pathways, crosstalk between autophagy and apoptosis, M1/M2 ratios, and specific microglial marker TLR4/NF-κB pathways are also important. Rapamycin and 5-aminoimidazole-4-carboxamide ribonucleotide are autophagy activators that can increase neuronal viability, reduce lactate dehydrogenase leakage, and alleviate apoptosis in oxygen–glucose deprivation/reoxygenation models by activating the adenosine monophosphate-activated protein kinase/DDiT4/mammalian target of rapamycin axis [37].

Nrf2 is a regulator of the antioxidant response at the cellular level and activates the expression of proteins and enzymes, which help neutralize ROS-induced ischemia–reperfusion injury. Nrf2 helps maintain mitochondrial function, biogenesis, and integrity, which are essential for neuronal survival during ischemia. Its activation can inhibit iron-dependent lipid peroxidation in the context of cerebral ischemia [38, 39].

In the global ischemic model, there is complete disruption of blood flow to the brain, leading to necrosis of the cerebral region. The methods used to achieve global cerebral ischemia include increasing intracranial pressure, occlusion of major arteries, and cervical compression [40]. However, a straightforward method for inducing ischemia is decapitation, which is performed after approval from the institutional animal ethics committee. This method was used long ago in small animals to elucidate the biochemical mechanism and global ischemic pathways involved. In the global cerebral ischemic model, any artery supplying the cerebral region can be occluded. The best example is middle cerebral artery occlusion in small-animal or large-animal model studies [41]. The middle cerebral artery occlusion model is widely used because it simulates thromboembolic stroke. Ischemic models can be generated in both small and large animals. Although many drawbacks exist in using animal models to study ischemia, these models are still in use.

There may be species-specific variations in neuroanatomy and neurophysiology, including responses to ischemia. The size and structure of the brain, vasculature, neuroprotection, and metabolic responses can vary among rat models, larger animals, and humans. Induction of cerebral ischemia by occluding the middle cerebral artery can produce variable results because of differences in surgical techniques, methods of anesthesia, and postoperative care. While animal model studies can yield good results, variable outcomes can cause difficulty in interpretation of the findings. Factors such as the genetic background of the animals can affect the severity of ischemic events and the recovery phase. Therefore, the findings of animal model research cannot be generalized. However, physiological control can be achieved in these models. With the help of these models, injury, its mechanism, and its neuroprotective effects can be better understood. These phases can be challenging to study in humans during ischemia because of the varied causes, manifestations, and ischemic sites. However, studying the mechanism of injury as well as neuroprotection is highly beneficial.

Nayak et al. [23] induced cerebral ischemia through bilateral carotid artery occlusion and thereby created oxidative stress. Ischemia results in a decline in memory and learning abilities; later, locomotor activity was assessed using an actophotometer, and motor coordination was assessed using the rotarod test. This was supported by assessment of cerebral infarction on the basis of histopathological findings. This was further supported by biochemical evidence of elevated lipid peroxidase (LPO) levels and depletion of SOD, CAT, reduced glutathione (GSH), and total thiols in the ischemic group. They also chemically induced ischemia by inducing oxidative stress through the administration of aluminum fluoride in drinking water for 7 days. There were decreased levels of SOD, CAT, and glutathione peroxidase (GPx), and elevated levels of oxidative stress markers, including malondialdehyde (MDA) and ROS.

The standard ICP threshold is more than 20 mmHg, and it can be measured with modern ICP monitoring devices, which may be invasive. Increased ICP can result from space-occupying lesions, cerebral edema, and hematomas. This reduces cerebral perfusion pressure and cerebral blood flow, leading to global ischemia and hypoxia. It causes mechanical distortion and compression of the brain, exacerbating ischemia [42]. A reduction in cerebral blood flow can lead to metabolic dysfunction and secondary brain injury. Research has suggested a significant decrease in cortical blood flow and velocity because of increased ICP [43]. In animal models of traumatic brain injury, increased ICP leads to increased neuronal injury and behavioral changes, even without global ischemia. This finding indicates that even subischemic increases in ICP can cause significant deficits [44].

Rodents, canines, pigs, and nonhuman primates have been used as animal models to induce increased ICP. Conditions such as an acute subdural hematoma cause increased ICP and exacerbation because of the mass effect of the hematoma. However, experimental animal studies often involve the infusion of artificial lumbar cerebrospinal fluid to increase ICP [43, 45]. In a Japanese study, autologous arterial blood was infused into the brainstem and internal capsule to increase ICP in white rabbits [46]. In another study, a Fogarty balloon catheter was inserted and inflated to simulate a space-occupying lesion and increased ICP [47].

Injection of autologous blood into the midbrain, internal capsule, and related sites can cause hemorrhage and increased ICP. Injection of saline into the lateral ventricle or brain can also cause increased ICP. In pigs, this method involves monitoring the optic nerve sheath diameter by ultrasound, which is an indirect indicator of increased ICP [48]. Direct measurement of ICP using transducers placed in the brain parenchyma or lateral ventricle is accurate but technically difficult [48].

Institutional animal ethics committee approval is mandatory to induce increased ICP in animals. It is also suggested that the induction and measurement of increased ICP can be complex due to the need for specialized equipment and expertise, potentially leading to variable results.

Cerebral ischemia can be experimentally induced through various arterial occlusions. The middle cerebral artery can be occluded by electrocoagulation through the application of an electric current to coagulate the artery. Devices such as filaments can be used and adjusted to create either permanent or transient occlusions of the middle cerebral artery. Application of the endothelin-1 peptide also induces vasospasm, leading to ischemia [49]. The common carotid artery can also be occluded to induce ischemia. Occlusion of the artery leads to decreased nutrition, reduced energy production, an acidic environment, and resultant cell death [50]. This method requires strict aseptic precautions and physiological monitoring under anesthesia. The limitation of this method is the presence of collateral circulation, which can significantly influence the outcome.

Middle cerebral artery occlusion models are commonly used in stroke research to simulate ischemic conditions in the brain. These models can be categorized into permanent and transient occlusion models. Permanent occlusion is usually induced by electrocoagulation, which permanently blocks the artery without reperfusion. Transient occlusion involves temporary occlusion of the artery with an intraluminal suture, followed by reperfusion after a period ranging from 30 min to 2 h. The Longa technique involves generating a focal ischemia model by occluding the right middle cerebral artery. The animals were excluded if a subarachnoid hemorrhage occurred during middle cerebral artery occlusion [51].

Given that ROS play a significant role in cell death following ischemic injury, several studies have been conducted to assess their neuroprotective effects. Modulation of hypoxia-inducible factor alpha has also been shown to increase endothelial nitric oxide synthetase and enhance cerebral blood supply in animal models of arterial occlusion [52]. Although this concept has shown positive results in animal models, in human trials these findings have not been as robust. This concept is successful in cardiovascular surgery but not in neurological disorders. The systematic use of imaging in ischemia and its management will allow us to understand the exact phenomenon occurring in the brain [53]. Therefore, there is a need to use combined approaches in ischemic conditions, including neuroprotective and vasoprotective agents, together with revascularization treatments.

The search for a natural neuroprotective substance with minimal side effects has attracted increasing attention. Although many plants have been examined to date, only a few have been studied thoroughly. Various compounds with antioxidant properties have been studied and tested to assess their endogenous antioxidant effects. Free radical scavengers and antioxidants are commonly used to mitigate the harmful effects of oxygen-free radicals during ischemia. There are potential clinical benefits, including reduced histopathological, biochemical, and oxidative metabolic damage, as well as improved neurological outcomes. A comparison of various drugs used to manage cerebral ischemia in animal models is presented in Table 2 [54–61].

Herbal products exhibit high variability in their medicinal value, particularly in antioxidant properties. This may be due to differences in species, the part of the plant used, climate conditions, and availability of these plants [62, 63]. Characterization and standardization of herbal products are based on various chemical constituents known as phytoconstituents. These phytoconstituents naturally occur in medicinal plants, leaves, seeds, vegetables, and roots and are responsible for multiple medicinal and pharmaceutical properties. They are classified into primary compounds, such as sugars and fats, and secondary compounds, such as alkaloids, tannins, terpenoids, phenols, and glycosides. These secondary compounds and other plant-derived chemical entities, such as Rheo discolor, are responsible for their medicinal value [64]. They have high contents of phenols (48.41 mg/1 g), flavonoids (43 mg/1 g), and tannins (44.93 mg/1 g). An earlier report found that phenols have strong antioxidant properties and reduce oxidative stress in the brain [65]. Flavonoids have been explored for their ability to prevent oxidative stress caused by ischemia–reperfusion. Tannins have also been reported to possess antioxidant and free-radical-scavenging activities [66]. These natural medicines are successful in preclinical studies because they are neuroprotective, ameliorating protein aggregation, oxidative stress, and neuroinflammation [67]. A comparison of the antioxidant content and strength of M. pruriens with those of other herbs is presented in Table 3 [68, 69].

Medicinal plants are gifts for fighting disease and death. Although many plants have been examined to date, only a handful have been studied thoroughly. The M. pruriens plant is also known as Kapikacchu, Atmagupta, and Naayi songe. This plant is easily digestible, has a high protein content, and is native to tropical regions of Africa, India, the Caribbean, and China. Its flowers may be white or purple. The pods have loose brownish hairs, which cause severe itching on contact with skin. The plant has black or brown beans. Currently, this plant is consumed in some countries because its pods can be used as a vegetable substitute for humans. In some parts of the world, its seeds are roasted and ground into a powder for use as a coffee substitute, such as Nescafé [70].

M. pruriens seeds contain approximately 27% protein and minerals. The antiphysiological and toxic properties of some M. pruriens species are due to the presence of amino acids, polyphenols, phytates, trypsin inhibitors, cyanogenic glycosides, oligosaccharides, lectins, saponins, and alkaloids. Polyphenols bind with proteins and thus decrease their digestibility [71]. In addition to levodopa, M. pruriens seeds also contain tryptamine and 5-hydroxytryptamine.

Its primary active compounds include levodopa, which is present at the highest level in its seeds, ranging from 0.2% to 7.30% of dry weight [72]. Its methanolic extracts yielded 3β-hydroxy-5α-cholanic acid acetate, 3,5,7,4-tetrahydroxy-6-methoxyflavone, and ethyl 2-amino-5-hydroxy-3,6,6-trimethyl heptonate. Other bioactive compounds, such as medicarpin and parvisoflavone B, have been identified as β-glucosidase inhibitors [73]. Ursolic acid and betulinic acid, which exhibit neuroprotective activities similar to those of levodopa [74], have also been detected. The medicinal properties of M. pruriens also include the production of alkaloids, including prurienidine, prurienine, and prurieninine [75]. (Z)-Triacont-5,7,9-triene, (Z)-docos-2,4,6-trien-1,8-diol, and (Z)-docos-5-en-1-oic acid are its lipid derivatives. Its secondary metabolites, such as flavonoids, tannins, saponins, steroids, glycosides, and terpenoids, have antimicrobial, antioxidant, and anti-inflammatory properties [76]. However, its antioxidant properties are affected mainly by phenols and flavonoids. The phytochemical profiles of M. pruriens are presented in Table 4 [77–80].

Levodopa reportedly enhances the ability to scavenge free radicals and reduce oxidative damage in cerebral ischemia [74]. It can cross the blood–brain barrier and restore dopaminergic tone. Flavonoids, polyphenols, and tannins scavenge nitric oxide, hydroxyl radicals, and superoxide. The levels of antioxidant enzymes, including CAT, SOD, and GPx, are increased, whereas MDA and lipid peroxidation are decreased. β-Sitosterol acts as a neuroprotective agent against cerebrovascular diseases by downregulating N-methyl-D-aspartate receptor gene expression and decreasing calcium influx, which prevents excitotoxic neuronal death [67]. β-Sitosterol is also involved in the suppression of ischemia-induced tau hyperphosphorylation and preservation of the integrity and stability of axons and microtubules.

M. pruriens is a traditional Indian medicinal plant that has been widely used in Ayurveda since the Vedic period. The plant is commonly known as Kapikacchu in Sanskrit and is used primarily for Parkinson’s disease and other nervous disorders, such as depression [81], and for arthritis, as well as for its aphrodisiac properties. In an animal model study with rotenone-intoxicated mice, M. pruriens and levodopa improved signs of Parkinson’s disease [82]. The n-propanol extract of boiled and fermented seeds of M. pruriens can offer greater neuroprotection against dopaminergic neurons than fresh seeds, as observed in a Parkinson’s disease rat model [83]. In other studies, the seed extract of M. pruriens enhanced learning and memory [84]. It increased the number of surviving neurons in the CA1 and CA3 regions of the hippocampus. Liver and kidney function tests revealed good recovery after long-term treatment with M. pruriens [85]. M. pruriens prevents depression-like behaviors by decreasing oxidative stress and lipopolysaccharide effects [86]. It has also been reported to have antiepileptic, antidiabetic, and antineoplastic effects [87].

The bioactive molecules of M. pruriens can be useful in the management of pulmonary hypertension [88]. The antivenom activity of this plant has also been investigated [89]. Levodopa is also responsible for other pharmacological effects of plants, such as antidiabetic, anti-inflammatory, neuroprotective, and antioxidant effects [90]. It is well known for its aphrodisiac properties, as it can increase sperm count and testosterone levels. Its use in infertility was confirmed in an animal model study in albino mice [91]. The seed extract of Mucuna decreased damage to tissue structure and sperm parameters in sodium arsenite-treated rats [92]. Mucuna contains levodopa, which helps in penile erection [93]. Its seeds have shown promising results in assays of total phenolic content, ferric reducing antioxidant power, and deoxyribonucleic acid damage protection [94]. A study from Thailand reported that M. pruriens seed extract prevented testicular apoptosis by affecting caspase, proliferating cell nuclear antigen, and dopamine receptor D2 protein expression [95]. During cell metabolism, free radicals are produced from one or more unpaired electrons. The free radical-scavenging properties of polyphenols make them important phytochemicals for managing oxidative stress. Flavonoids are simple phenolic compounds with antioxidant properties [96]. In experimental rodents, the ethanol extract of M. pruriens leaves was shown to exert anti-inflammatory activity by inhibiting oxidative stress, proinflammatory cytokines, and lysosomal membrane instability [97].

M. pruriens is helpful against global cerebral ischemia/reperfusion injury-induced oxidative stress in rats, and it is likely to be useful in ischemic treatment. The methanolic extract of M. pruriens was shown to reduce the incidence of ischemic brain injury-induced by occlusion of the common carotid artery in a rat model [98]. Nayak et al. [23] reported that M. pruriens ameliorated decreased activity observed in ischemic animals. This treatment decreased the elevated LPO level and increased the levels of SOD, CAT, GSH, and total thiols in the ischemic group, suggesting a protective effect of the extract against oxidative stress. In this study, histopathological observation of hippocampal CA1 neurons revealed a decrease in the number of viable neurons in the stress-induced group. Animals in the M. pruriens extract-pretreated ischemic group presented a dose-dependent decrease in neuronal damage, indicating neuroprotective activity. The observed impaired motor coordination in the aluminum fluoride-treated animals was minimal in the extract-treated groups. M. pruriens can alter neurotransmitter levels and the balance of excitatory and inhibitory inputs to Purkinje cells of the cerebellum.

M. pruriens offered neuroprotection in a global cerebral ischemia animal model induced by occlusion of the bilateral common carotid artery [23]. This was evident in histopathological and biochemical analyses. It was reported that treatment with methanolic extracts of M. pruriens seeds improved locomotion, coordination, and spatial learning. It also reduced the cerebral infarct area, indicating its therapeutic efficacy in treating cerebral ischemia [99]. This is due to its antioxidant properties, which increase the levels of antioxidant enzymes and reduce lipid peroxidation [100]. A methanolic extract of M. pruriens significantly increased the levels of SOD, CAT, GPx, and glutathione reductase in high-fat diet-fed rats [101]. It also elevates nonenzymatic antioxidants, such as glutathione, further indicating a reduction in oxidative stress. This study also revealed a decrease in thiobarbituric acid-reactive substances, which are markers of lipid peroxidation, and this finding was statistically significant (p < 0.05). Krishna and Sundararajan [102] compared the effects of different doses of the methanolic extract of M. pruriens and reported that higher doses significantly increased the ferric reducing ability of plasma and CAT levels (p < 0.05). It also reduced thiobarbituric acid-reactive substances levels, suggesting a dose-dependent antioxidant effect of M. pruriens. It has been reported that prophylactic treatment with M. pruriens and its bioactive molecule, β-sitosterol, downregulates the expression of the N-methyl-D-aspartate receptor and tau protein genes in the ischemic brain, which are usually upregulated during cerebral ischemia [98]. The comprehensive mechanism of action and neuroprotective role of M. pruriens are schematically represented in Figure 2 [21–26, 45, 70–87, 91, 94, 97]. M. pruriens has potential for preischemic conditioning, which is similar to ischemic preconditioning. It is hypothesized that pretreatment with M. pruriens may enhance brain resilience.

M. pruriens has been proven to be neuroprotective in ischemia-induced animal models, but its efficacy can vary. Its effect is due to antioxidative and anti-inflammatory properties; however, this effect may not be sufficient in all cases of cerebral ischemia [23, 99]. In a spinal cord injury animal model, no statistically significant beneficial effects were observed, suggesting variable efficacy across different types of neuronal injury [21]. Another challenge is its bioavailability, as the blood–brain barrier can limit delivery of Mucuna compounds to the brain, potentially decreasing their efficacy in treating cerebral ischemia. The dosage and route of administration of M. pruriens need to be standardized to ensure that active compounds reach brain tissue.

The application of translational research and results from animal model studies to human clinical trials and experimental setups may be uncertain. Variability among individual subjects and efficacy across various species present significant challenges. There is a need for better guidelines to achieve significant therapeutic outcomes [23, 98, 99]. M. pruriens is effective as a potential neuroprotective agent because of its relatively high content of levodopa, which is a precursor of dopamine, and other bioactive compounds, such as tyrosine [86]. Hence, M. pruriens can be contraindicated in patients with comorbidities such as Parkinson’s disease, where it is already used as treatment, because it can cause toxicity [103]. The quantity of bioactive compounds in M. pruriens can vary depending on the extract and preparation methods, which can lead to variability in efficacy [90]. Long-term use of M. pruriens can cause levodopa-like dyskinesias [104]. Hence, understanding these side effects in future studies is crucial.

It has also been reported that unprocessed M. pruriens may contain antinutritional components and could be toxic to ruminants [76]. In some regions, such as Japan, M. pruriens products are not pharmaceuticals and can be sold online, and permission from food regulatory authorities is not needed. However, specific legal requirements apply, and these products should not be advertised inappropriately, as this may lead to incorrect messaging and misuse [105].

Although M. pruriens effectively reduces ischemic damage to the brain, as evidenced by histopathological analysis, the degree of neuroprotection can vary. Further research is essential for understanding the potential factors responsible for this variability [98]. The present evidence comes from preclinical animal model studies, and human clinical trials are essential for establishing standardized therapeutic guidelines. More research on M. pruriens is needed to study its comprehensive effects in treating cerebral ischemia, as its etiopathogenesis is complex, involving oxidative stress, neuroinflammation, and neuronal death. The future implications include studying it as an adjuvant and as a combination therapy along with other drugs. Nanoformulations of M. pruriens, blood–brain barrier-permeable phytochemical fractions, standardized extraction methods, and combination therapy with reperfusion drugs should be studied in future research.