Study the effect of Malva sylvestris leaf extract (MSE) against CCl4-induced nephrotoxicity in a model of male rats

Introduction

The kidneys, one of the most important organs in the body, are involved in various activities. Some of their activities include the production of urine that is removal of waste from the bloodstream is considered the primary role of the kidney most physicians refer to it as producing urine [1] . It aids in maintaining volume, pH, and ionic balance, among other homeostatic activities. In addition, other harmful metabolic byproducts are expelled [2] . Nephrotoxicity refers to the manifestation of kidney damage after exposure to drugs, chemicals, or other toxic agents. It is a severe field of concern since it is associated with acute renal failure, which has a significant impact on overall health [3] . Oxidative stress is in most cases implicated in the mechanisms of nephrotoxicity since it plays a vital role in the injury of renal cells caused by a range of nephrotoxic agents such as heavy metals and industrial chemicals and aminoglycoside antibiotics [4] . The first well-investigated nephrotoxin is CCl4. CCl4 acts with the first biochemical response which leads to the biochemical backlash of the cytochrome P450 in a hepatocyte. High reactivity causes injury post the development of acyl-CoA products. The injury developed through the down regulation of enzyme replication, and it initiates cellular apoptosis [5] . Malva sylvestris, commonly known as common mallow, is native to southern Europe and western Asia. M sylvestris is a biennial-to-perennial herbaceous plant from the Malvaceae family [6] .

Malva, is a tall herb with broad, heart-shaped, light green leaves that grows 2-3 feet in height. Beautiful purple blossoms that appear to be honeysuckle are produced by the plant. Malva flowers have been bioengineered with a thresher and are used for ME, which have been implicated in creating pharmaceutical agents authentic to microbial infections, colds, trachea, bronchi, beef cavities, salmonella, and general cardiac quarantine [7] . Several pharmacological activities have been conducted on Malva sylvestris L. because it possesses diuretic, laxative, expectorant and antitussive traits and is used to treat organ disorders such as diseases of the respiratory and digestive systems. The phytochemicals in Malva sylvestris cause it to act as an antioxidant because it counters oxidative stressors and prevents various illnesses, including CKD and diabetes [8] . Because of a wide range of nutrients such as calcium, potassium, magnesium, iron, and vitamins, Malva has been distinguished as a medication for a broad range of disorders [9] . Although traditionally the leaves, flowers, and seeds of the Malva sylvestris plant have been used as a healthy food in addition to a dietary problem, they are not as commonly consumed as before. The seeds taste like young hazelnut, while the leaves and young flowers were added to salads [10] . Due to the high quality of the mucilage stock, mallow is frequently used in folk medicine, and their use in other treatments is very common for a variety of reasons, such as for stomatitis and asthmatic bronchitis. Apart from that, Malva sylvestris is well-known for its antiinflammatory, antibacterial, and astringent properties and can be used in the effective treatment of acne, carbuncles, furuncles, insect bites, and eczema, to name but a few [11,12] . This study was conducted to examine the model of nephroprotection by Malva sylvestris leaves on CCl4-induced renal damage in the rat.

Materials and Methods

Plant Material and Extract: Preparation Leaves of Malva sylvestris L. were collected and authenticated from Babylon province, Iraq. The plant leaves were air-dried under shade in ambient temperature for two weeks. After drying, the plant material was grinded and sieved through a fine mesh sieve to obtain the fine powder . The hydro-alcoholic extract of MSE was prepared by macerating Eq. 400 gram powdered leaves in Eq. 800 mL of 70% ethanol for 72 hours. After 72 hours, the extract was filtered and the solvent was evaporated under reduced pressure in the rotary evaporator to get the methanolic extract. The crude methanolic extract was stored at 4 °C in the Refrigerator until further use.

Experimental Animals: Healthy adult male Wistar rats, weighing 180-220 g acclimatized for a period of 7 days before starting orally fed with minimum acclimatization period of 1 week. Animals were kept under controlled condition and maintained under 22±2 °C, 12 hour light/dark cycle at 50-60% humidity. The rats were allowed for food ad libitum and water ad libitum. The experiment was performed after getting approval from the Institutional Animal Ethics Committee in Al-Qasim green university and complied with the national laws related to animal care and use. Oral treatments commenced one week prior to the first CCl4 administration and continued throughout the four-week CCl4 exposure period. As shown in Figure 2, kidney sections from the CCl4-treated group had significant histopathological disturbances in the cortex and medulla with tubular necrosis, vacuolar degeneration of epithelial cells, and an extremely dense inflammatory cell presence indicative of acute renal injury. In marked contrast, kidney sections from the MSE-treated group maintained the normal histology of renal tissue, and minimal inflammatory cellular background was seen which suggests that the MSE possessed a protective effect against CCl4induced nephrotoxicity. The MSE-treated and NC groups showed approximately normal histologic architecture with minor changes. The MSE possessed nephroprotective activity due to its anti-inflammatory and antioxidant efficacy, which is in accordance with the biochemical examinations.

Experimental Design

Results

As shown in

Discussion

In this experiment, the protective impact of Malva sylvestris leaf extract on CCl₄-induced nephrotoxicity in male rats was investigated. The results revealed that CCl₄-induced toxicity cause significant renal dysfunction, as exhibited by elevated BUN and serum creatinine, increased lipid peroxidation, and decreased activity of renal endogenous enzymatic and nonenzymatic antioxidant defense mechanisms like SOD, CAT, and GSH levels [13] . It is consistent with past findings that CCl₄-induced nephrotoxicity by the generation of ROS and trichloromethyl radicals and thus causing lipid peroxidation and oxidative damage. Overall, administration of MSE markedly reversed these derangements in a dose-dependent manner, indicating an antioxidant-mediated nephroprotective effect.

The extract particularly at 200 mg/kg produced a remarkable reduction in renal biomarkers, MDA concentration, and an increase in the levels SOD, CAT, and the GSH. It demonstrates that MSE could prevent oxidative stress and enhance the ability of the kidney antioxidant defense system to reduce cellular damages by free radicals [14] . The antiradical activity of M. sylvestris can be justified by its remarkable amounts of flavonoids and phenolic acids and polysaccharides, which demonstrate radical scavenging in addition to metal binding activities. Similar antioxidant and somatoprotective effects of M. sylvestris have been confirmed in hepatic and renal toxicity models [15] . MSE bioactive components like quercetin, kaempferol, and gallic acid might have contributed to the stabilization of cellular membrane integrity and thus the conservation of enzymatic tissues antioxidant defense systems. Furthermore, it was found to cause a significant decrease in lipid peroxidation, representing an inhibition of the peroxyl radical chain during reduction within the renal tissues [16] . The increased level of SOD and CAT shows the activation and protection of the two critical antioxidant enzymes against inactivation by oxidation by MSE administrations. Moreover, the raised content in GSH revealed a reinvigorated redox balance because it detoxifies hydrogen peroxide and lipid hydroperoxides [17] . Therefore, all parameters suggest that the extract could reinstate the oxidative balance in the treated kidney. These results supported the previous reports that natural antioxidants may provide substantial nephroprotection from chemically induced oxidative damage. CCl₄-mediated nephrotoxicity has been studied as a model of oxidative stress-mediated nephropathy [18, 19] . This structural confirmation of MSE's protective effect was extended by histopathological analysis. In the CCl4 group, the kidneys presented severe architectural damage, marked by tubular and outer medulla necrosis, and inflammatory cell infiltration, indicative of acute renal injury. It is clear that administration of MSE before CCl4 exposure significantly reduced the severity of lesions observed in the CCl4 groups. Renal tubular and glomerular architecture was relatively well preserved and differences were observed. These structural improvements reinforced the biochemical findings and suggested that MSE protect cellular and tissue integrity when imitating toxic attack. The reduction in the entrance of inflammatory cells into the kidney may be due to the anti-inflammatory impact of MSE, whether direct or indirect. MSE may also reduce inflammatory cell filtration by reducing the expression of cytokines due to its antiinflammatory effect. The fact that MSE protects the nephron from oxidative attack means that it can be used as a supplement or prophylactic measure for the treatment of kidney oxidative pathologies.

Conclusion

To conclude, it can be inferred that Malva sylvestris leaf extract reversed CCl₄-induced oxidative renal damage, and it exerted a marked nephroprotective and antioxidant effect against rats. It substantially restored the most quantitatively important renal function and levels of activity of antioxidant enzymes and diminished the levels of lipid peroxidation in a dose-dependent effect of CCL4. The results further imply that M. sylvestris has an ability to attenuate oxidative stress via the enhancement of endogenous antioxidants generated by antioxidants and preserving renal cellular integrity. This conclusion suggests that it can be used in future studies to isolate active ingredients, or even complete analysis of the molecular pathway as a nephroprotective remedy.

Conflict of Interest Not available

Financial Support

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