The E. Prostrata plant is traditionally used by local people to treat various ailments, depending on several bioactive substances that are naturally present in the plant to maintain their health. The extraction procedure aims to efficiently isolate a maximum number of target compounds from plant extracts while preserving and maximizing their biological activities. The extraction method and extraction solvent influence the extraction yield and biological activity of the extract [38, 39]. The best solvent for the extraction of phytochemicals from plants is methanol due to its high polarity and low boiling point of just 65 ⸰C. Soxhlet apparatus is used to extract the pure bioactive compounds from the plant material [40]. It may be inferred from the preceding discussion that the best solvent to extract phytoconstituents from E. prostrata is methanol.
In this study, DPPH scavenging activity and ABTS decolorization-reducing power tests were performed to estimate the antioxidant properties of the methanolic extract made from E. prostrata. According to our research, the methanolic extract of E. prostrata displayed strong antioxidant activities, which were demonstrated by its outstanding results in the DPPH and ABTS tests. The extract significantly high concentration of bioactive compounds may be what causes this increased antioxidant activity. As a result, our findings concur with those of earlier research that has described comparable effects [41]. These phytochemicals remarkable capacity to successfully neutralize a variety of ROS confers protection against the damaging effects of oxidative stress. These findings imply that the E. prostrata methanolic extract would be a suitable antioxidant option for pharmaceutical development. We started an experimental examination to establish the effectiveness of E. prostrata because it has historically been used to treat a variety of illnesses. In some earlier investigations, the anti-hyperlipidemic effects of E. prostrata were studied in rats that had high-fat diet-induced hyperlipidemia [19]. In this experiment, we determine the effects of the methanolic extract from E. prostrata in acute diabetes mice models. Notably, the methanolic plant extract from E. prostrata significantly lowered glucose levels in acutely diabetic mice models, and its effectiveness was comparable to that of the artificial medication glibenclamide.
Alloxan leads to chemical diabetes by damaging the pancreatic cells that release insulin. As a consequence of the many cells that are harmed during this process, less insulin is generated and less glucose is absorbed by the tissue. Glibenclamide belongs to the class of drugs called sulfonylureas. Glibenclamide lowers blood glucose levels by increasing the insulin your pancreas produces [42]. These phytochemicals might encourage insulin production from the surviving islet cells, which could be the possible mechanism behind the anti-diabetic actions of the E. prostrata extract. Some bioactive compounds help to boost the sensitivity of tissues of the body to insulin. The amount of blood glucose that cells absorb is regulated by the insulin hormone. Possibly different bioactive compounds from the methanolic extract of E. prostrata improve insulin sensitivity, which helps to reduce blood sugar levels. Recent studies demonstrated that E. prostrata contains some bioactive compounds that are good in their anti-diabetic potentials [43, 44].
Alkaloids may unite to the competitive or noncompetitive sites of enzymes mixed up in digestion, preventing the creation of an enzyme-substrate complex and, as a result, reducing enzyme activity [45]. Several alkaloids consequential from E. prostrata have been shown to improve glucose absorption. Vindolicine III was shown to engage in glucose absorption in C2C12 and β-TC6 cells, as well as having more antioxidant effect than vindolinine IV, vindolidine II, vindoline I, and making it effective in curing hyperglycemia [46]. Oxidative stress carried on by hyperglycemia causes both micro and macrovascular issues. Diabetes problems can be reduced by the treatment of antioxidants. Antioxidants act by donating and accepting electrons, or by reducing free radical production by inhibiting the enzyme expressions and activities, responsible for antioxidant generation [47]. Flavonoids inhibited the activity of aldose reductase and α-glucosidase [48]. Anthocyanins are crucial polyphenols that are found in E. prostrata. They are investigated and found effective in decreasing obesity and treating type-2 diabetes mellitus. Moreover, these compounds affect glucose absorption, lipid metabolism, and insulin secretion [49, 50]. Furthermore, anthocyanins retarded the intestinal absorption of glucose, decreasing the blood glucose level [51, 52]. The cyaniding-3-galactoside is a natural anthocyanin. This compound was investigated for the inhibitory action of α-glucosidase in the intestine. It also showed synergistic inhibitory activity. These studies suggest the inhibitory activity of cyaniding on intestinal α-glucosidase and pancreatic α-amylase enzymes [53, 54].
Phytochemicals provide large-scale research in anti-depressant therapy. Alkaloids, amines, saponins and sapogenins, terpene and terpenoids, and carbohydrates containing plant metabolites were shown to have anti-depressant effects. Polyphenols (flavonoids, phenolic acids, lignans, and coumarins) were another group of plant metabolites that showed anti-depressant function. The diterpene alkaloids songorine, napeline, hypaconitine, and mesaconitine increased serotonergic system activity in depressive animals [55]. The modulatory effects of evodamine on BDNF-TrkB signal and monoamine transmitters in the hippocampal regions may be involved in the causative process [56]. Piperine increases serotonin levels in the limbic and cerebral cortices, producing antidepressant-like effects [57]. Anti-depressants are drugs that are often used to treat depression and other mental health issues. They function by specifically targeting and altering the amounts of neurotransmitters in the serotonin, brain, norepinephrine, and dopamine [58]. Both the tail suspension and forced swim experiments put mice in an unavoidably stressful environment, which led to a state of despair and hopelessness that may be explained by the length of immobility. Immobility or despair behaviour is induced in both tail suspension and forced swim experiments so this behaviour is used as a model for depression. Administration of therapies for depression and anti-depressants reduces the immobility time. Methanolic extract of E. prostrata reduced the immobility time in the forced swim and tail suspension test indicating an antidepressant-like effect. Our results align with many studies [59]. The sucrose preference test was for the assessment of depressive behaviour. Depressed mice lost their pleasure. Glibenclamide and crude extract of E. prostrata increased the sucrose preference. Both low dose (150mg/kg) and high dose (300 mg/kg) of methanolic extract of E. prostrata significantly boost the sucrose intake; it means that methanolic extract of E. prostrata possesses anti-depressive activity. This study results line up with some earlier studies [60, 61].
A neurotransmitter called serotonin is essential for scheming mood, emotions, and general comfort. It often evokes emotions of joy, relaxation, and fulfillment. Serotonin plays an important role in comprehending the underlying cause of depression. Low serotonin levels and other neurotransmitter levels impair regular brain transmission, which results in the emergence of depressive symptoms. Alloxan monohydrate decreases the serotonin level in the brain. After the treatment of the treated group with plant extract and glibenclamide, serotonin level was increased as compared to the untreated diabetic group which indicates that plant extract possesses an antidepressant and anti-anxiety effect. This study aligns with an earlier study [62]. TSH, or thyroid-stimulating hormone generated in the brain by the pituitary gland. Its principal function is to control thyroid hormone synthesis and release from the thyroid gland. Our results demonstrated that no medication had a significant impact on TSH levels in the brain.
It was discovered through phytochemical analysis that 55 key bioactive compounds were in charge of controlling neurotransmission to produce anxiolytic effects. The five chemical families that make up these bioactive ingredients are as follows: alkaloids, polyphenols, terpenoids, phytosterols, and others such as fatty acids and miscellaneous. Alkaloids can alleviate anxiety by acting on the glutamatergic neurotransmitter system. Anxiety occurs by releasing acetylcholine in the critical regions of the brain like the ventral hippocampus. Postsynaptic and presynaptic sites of neuron cells have two main types ionotropic, nicotinic, and metabotropic muscarinic receptors of acetylcholine. Positive charges most probably Ca2 + allowed to flux into the cell and as a result, it depolarizes the membrane of neurons [63]. It is due to acetylcholine when it is released from pre-synaptic neurons. An increase in acetylcholine level may cause emotional irregularity like anxiety. Anxiety may arise when the enzyme acetylcholinesterase (AchE), responsible for breaking down acetylcholine, is inhibited. So flavonoids inhibited the activity of AchE enzyme [64].
Mice with diabetes spent much less time in the inner zone or the center of the box as compared to the control group. Furthermore, compared to the diabetic group, the injection of E. prostrata might dramatically lengthen the inner zone. Compared to control mice, diabetic mice spend much more time at the outside or perimeter of the box. Additionally, compared to the diabetic group, the injection of E. prostrata might lower this score [21]. There was a significant down in the exploration time of novel objects in the testing period in diabetic mice. When compared with the diabetic group, the glibenclamide and plant crude extract treated group increased the exploration time of novel objects [65]. The mice in the diabetic group that received alloxan monohydrate showed a significant increase in the time spent in closed arms and a significant decrease in the time spent in open arms in comparison to the control group, according to the findings from the EPM test. E. prostrata, on the other hand, showed a marked increase in time in the arm that was open and a considerable reduction in time spent in the closed arm in the diabetic and all treated groups. Effective results from these tests may maybe due to the presence of some phytochemicals in plant extract from already mentioned Anti-diabetic, anti-depressive, and anxiolytic phytochemicals. The results of this study were aligned with some earlier studies [21, 66]. Diabetes, depression, and anxiety have a complicated and bidirectional association. Diabetes patients have a greater rate of depression than the overall population, according to studies. Individuals suffering from depression are also more likely to get diabetes. Diabetes and depression share multiple risk factors, including genetic susceptibility, being obese, physical inactivity, and poor lifestyle choices. Furthermore, both illnesses have been linked to chronic inflammation and hormone abnormalities, which may contribute to their coexistence. Diabetes may change the chemistry and function of the brain, thus raising the chance of developing depression. Diabetes-related blood sugar changes, insulin resistance, and chronic inflammation may all impact neurotransmitter function and contribute to mood problems. According to the results of our study, diabetes, and depression have a relationship with each other. Changes in the serotonin level of the mice with diabetes and the mice treated with plant extract support our study. According to the findings of our study, diabetes, depression, and anxiety had a relationship with each other. Diabetes, depression, and anxiety are connected health conditions that interact in a complex way. Both those with diabetes and those who already have these diseases are more prone to develop sadness and anxiety.