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B Sirisha^1,2 , D Meera¹ , Ch. Venkatramana Devi¹

¹Department of Biochemistry, Osmania University, Hyderabad-500007, Telangana, India

² Telangana Social Welfare Residential Degree College for Women, Mahendrahills, Hyderabad, Telangana, India

Corresponding Author Email: boyinisirisha24@gmail.com

DOI : https://doi.org/10.51470/APR.2024.03.02.18

Abstract

Keywords

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1. Introduction

According to the WHO, 21,000 plants reutilized for medicinal purposes worldwide, and India has a rich tradition of using medicinal herbs and spices. There are 2,500 species in India, while 150 of them are used extensively for economic purposes[1]. Day-by-day usage for plant-based medications is rising globally including in India[2].Due to the presence of phytochemicals, most of the plants show healing properties to many diseases and showing anti-bacterial, characteristics which include antifungal, anticancer, antidiuretic, anti-inflammatory, along with anti-diabetic[3-7]. Presently many countries are looking for analternative to allopathic treatment for many diseases like diabetes, cancers, and bacterial and viral infections. Medicinal plants are rich sources of components that are used in the production of medication. There are numerous pharmaceutical companies that specialize on producingmedicationsusing plants as raw materials [8]. Due to the extraction of active compounds used in the production of different medications; medicinal plant raw materials are frequently used. Similar to laxatives, blood thinners, antibiotics, or antimalarial medications, all of these medications contain plant-based components[9].

Nanoparticles synthesis of silver and gold, its usage in the numerous advancements in the area of cancer diagnostics and treatment via nanoparticles (NPs), and medicinal applications have constantly increased[10]. Due to the demanding usage of nanoparticles, they are synthesized using hazardous substances with both physical and chemical techniques. To overcome this, many researchers are focused on the synthesis of eco-friendly nanoparticles from enzyme[11], fungus[12], and algae[13-14] were reported successfully in the production of Au-NPs. Additional to this, synthesizing the nanoparticles from plants and plant parts and its usage in diseases management are very effective compare to the plants alone[15].

Achyranthes aspera Linn is a plant species. belongs to the Amaranthaceae family, which is also known as Apamarga. It is commonly found as a weed in India. Furthermore, it is used as traditional medicine in tropical Asian and African countries[16].With fixed branches, thick, elliptic leaves, along with greenish-white blooms, it can reach a height from 0.3 – 1 m.Antibacterial, antifungal, thyroid-stimulating, antiperoxidative, anti-inflammatory, antiarthritic, immunomodulatory, wound-healing, anti-obesity, anticonvulsant, anticancer, along with hepatoprotective qualities had been among the various biological qualities demonstrated by A. aspera. Numerous phytochemical classes, including alkaloids, steroids, flavonoids, phenolic compounds, saponins, orterpenoidshavebeenreported to occur in this plant.

• Material and Methods

2.1 Collection of the plant material

Achyranthesasperaplantswere procured locally, with Dr. Shashikanth, a taxonomist of the Department of Botany at Osmania University in Hyderabad, verifying it. Following plant harvest, the surface was carefully cleaned with running tap water to remove if any remaining dust particles of the plant separating the leaves, root and stems from the plant. Separated plant parts leaves, root and stems were shade dried and ground separately into powder forfurther experimental analysis.

2.2 Preparation of Extracts

25g of dry coarsely powdered plant material (Root, Leaf, and Stem) separately was submerged in a 250ml of double distilled. The mixture was heated for 20minutes at 60°C while stirring occasionally. The mixture is cooled to room temperature and filtered using Whatman no.1 filter paper. The filtered extracts (Root, Leaf, and Stem) separately were stored at 4°Cfor further experimental analysis[26].

• Synthesis of Silver nanoparticles (AgNPs)

A. aspera leaf, stem, and root extract (10 mL) were added to 90 mL containing 1 mM aqueous silver nitrate solution, which was then heated to 80 °C for three hours while being constantly stirred in order to create silver nanoparticles. The shift to yellow to a rich brown coloration indicated the production of AgNPs. Labeling was done on the final green-synthesized silver nanoparticles, which then remained at 4°C until additional experimental examination[27].

2.4Synthesis of Copper nanoparticles (CuNPs)

A. aspera copper nanoparticles have been made by mixing 50 mL (5 mM) copper sulfate solution with 5 mL of aqueous plant extracts of the leaf, stem, along with root. NaOH (1 N) solution had been added to lower the mixture’s pH to 7.0. The resultant green mixture was additionally stored for further experimental analysis at room temperature[28].

2.5Synthesis of bimetallic Silver-Copper nanoparticles (Ag-CuNPs)

A. aspera bimetallic silver-copper nanoparticles have been established by mixing equal volumes of nanoparticles of silver and copper (50:50) and holding them at 4°C until additional experimental investigation.

2.6 In Vitro – Hepatoprotective Activity

HepG2 cells were used to examine the test extract’s in-vitro hepatoprotective properties. The toxicological assay involved incubating cells with DMEM in DMSO (0.05% v/v) for 12 hours, followed by 1.5 hours of DMEM treatment with 40 mM CCl₄. Normal control cells were cultured with DMEM in DMSO (0.05% v/v) for 12 hours. Cells were treated in 40 mM CCl₄for 1.5 hours after being cultured with DMEM containing various extracts at concentrations of 10, 25, 50, 100, and 150 μg/mL for 12 hours. By reducing the MTT salt to chromophore formazan crystals, the cells with metabolically active mitochondria produce precipitates at the end of the incubation time. For solubilized crystals with DMSO, the optical density is evaluated at 570 nm using a microplate reader. The following formula has been employed to determine the growth inhibition percentage.

• Results

Silver, copper and silver-copper combined Nanoparticles synthesized from the Acyranthusaspera root, leaf and stem extracts and have been studied for thierin-vitrohepato-protective activity using the HepG₂ cells. HepG₂ cells treated with CCl₄(40mM)alone and CCL₄ along with Root extract and Root silver, copper and Silver-Copper nanoparticles for the concentrations of 10-150 µg/ml.CCL₄alone showed the inhibition of 52.55±0.526 at the concentration of 40mMCCl₄. A combination of CCL₄(40mM)and  Root extract treated with the 150 µg/ml showed an inhibition of  33.1±0.256 whereas the Root Silver-Copper nanoparticles showed an inhibition with 24.57±0.327followed by Root Copper nanoparticles with the inhibition 25.25±0.268 and Root Silver nanoparticles with 27.3±0.325 of inhibition, 

A comparison of the Hepatoprotective activity of Root, Leaf, and Stem Ag-Cu A.aspera Extracts has shown that the highest concentration of Stem Ag-Cu extract has shown a significant increase inthepercentage of Inhibition as compared to Root and Leaf.

• Discussion

Hepatotoxic chemicals impact liver cells by inducing oxidative damage, or the liver is crucial for the metabolism along with detoxification of substances that into the body which have the potential to induce hepatic injury.Both synthetic as well as natural medications are available to treat liver disease.Liver disorders have traditionally been treated with natural therapies, and in recent years, plant-based herbal medications have become increasingly important in the fight against drug-induced toxicity[18].

The in-vitro hepatoprotective efficacy of gold nanoparticles made from Moringa oleifera pods was investigated; Likewise, ethanolic, chloroform, and aqueous extracts revealed that M. oleifera might have hepatoprotective properties[19].Aqueous leaf extract from Azimatetracantha has been employed to generate silver nanoparticles, which demonstrated potent hepatoprotective efficacy against CCl₄-induced hepatocyte damage[20].In our present study,acombination of Silver-Coppernanoparticles synthesized from the Acyranthusaspera leaf extract showed highest heapato-protective activity compared to the silver. A study on the hepatoprotective and antioxidant effects of cortex dictamni aqueous extract in rat liver damage caused by carbon tetrachloride (CCl₄) revealed that rats of both sexes with CCl₄-induced liver damage had noticeably low levels of aspartate aminotransferase, alkaline phosphatase, glutamate pyruvate transaminase, and total bilirubin[21].Clerodendrumpaniculatum Flower Extract showed better hepatoprotective activity in CCl₄-induced hepatotoxicity due to the isolated fraction containing the rich flavanoidsin female Wistar Albino rats[21].

Green silver nanoparticles from Herpetospermumdarjeelingense were produced, and it was found that these particles improved hepatoprotective function in addition to antimicrobial and antioxidant activity[23].Nanoparticle (AgNPs) synthesized from the ethanol extract showed in vitro hepatoprotective efficacy along with resistance to paracetamol-induced hepatotoxicity in rats evaluated for Broussonetiapapyrifera L., Alangiumsalvifolium, while Abutilon indicum L. AgNPs produced by Broussonetiapapyrifera L., Alangiumsalvifolium, along with Abutilon indicum L. shown notable hepatoprotective effect by lowering the metabolic parameters altered by CCL₄ along with paracetamol[24].

Methanolic leaf extractofAcalyphaindica, silymarin and quercetinredcued the toxicity CCl₄ induction by the antioxidant and anti-inflammatory properties since the extract contains flavonoids[25]. In our present study, we have synthesized the silver, copper and combination of silver-copper NPs from theAcyranthusasperaleaf,root and stem extracts and evaluated the in vitro hepatoprotective activity against the CCl₄ induced toxicity in HepG2cells.Thecombination of silver-copper nanoparticles of stem showed the better hepatoprotective activity as compared to silver-copper nanoparticles from leaf and root.

• Conclusion

The combination of silver-copper nanoparticles from leaf, root and stem showed the hepatoprotective activity compared to the individual nanoprticles of silver and copper nanoparticles form Acyranthusasperaleaf, root and stem extracted nanoparticles. Combination of silver-copper nanoparticles from the stem showed a prominent hepatoprotective activity than the combination of leaf and root-derived silver-copper nanoparticles. According to our research, silver-copper nanoparticles generated from A.asperastem have the capability to serve as protective agents of liver against damage caused by carbon tetrachloride. Further investigations are warranted to confirm our studies.

Credit authorship contribution statement

B SirishaWriting–original draft, Validation, Methodology, Investigation, Conceptualization.ChVenkataramana Devi: Supervising – original draft, Project administration, Conceptualization.

Declaration of competing interest

We, the authors declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

The authors are thankful totheDepartment of Biochemistry and Botany, University College of Science, Osmania University, Hyderabad for providing infrastructure and necessary support.

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