\u00a0Materials and Methods<\/strong><\/p>\n Collection of plant and authentication<\/strong><\/p>\n Gliricidia sepium <\/em>plant specimens were obtained from the districts of Sangli and Kolhapur, and they were further verified by the Botanical Survey of India (BSI) in Pune, with certificate reference number NO. BSI\/WRC\/100-1\/Tech.\/2020\/128 dated 28\/01\/2021.<\/p>\n Preparation of plant extract<\/strong><\/p>\n \u00a0The collected aerial parts were washed and then dried under shade.\u00a0 The plant material was subjected to crushing, resulting in the production of coarsely powdered particles. The coarse particles were subsequently subjected to sequential extraction using chloroform, and ethanol solvents by using Soxhlet apparatus. 7, 8<\/sup><\/p>\n Animal Experimentation<\/strong><\/p>\n Female Wistar rats were used for all the experimental procedure and were obtained from animal house of Appasaheb Birnale college of pharmacy, sangli (Approved from CCSEA of Appasaheb Birnale college of pharmacy, sangli IAEC no: IAEC\/ABCP\/18\/2019-20) at 45th<\/sup> days of age weighing 150 \u2013 200 gm. The animals were kept in a 12-hour light\/dark cycle with typical laboratory settings. They were given food and water ad libitum. 42 animals were divided randomly into seven groups (N=6) N-methyl-N- nitrosourea (50 mg\/kg) dissolved in isotonic saline solution. The first dose of NMU injected by IP route on 45th<\/sup> \u00a0day postnatal day, second one on the 55th<\/sup> \u00a0day and 70th<\/sup> day postnatal day drug while standard drug tamoxifen, chloroform extract of Gliricidia sepium <\/em>at dose of 200mg\/kg (CHGS 200) and ethanol extract of Gliricidia sepium <\/em>at dose of 200mg\/kg (ETGS 200) were used as test compound and is administered from 1st<\/sup> \u00a0day of model after end of study model animal will be sacrificed for biochemical and Histopathological studies 9,10,11<\/sup><\/p>\n Histopathology<\/strong><\/p>\n Thin tissue slices (3\u20135 mm) were extracted from both normal and pathologically altered tissues. The samples were fixed in 10% formalin at room temperature for 24 to 48 hours. After fixation, the tissues were deparaffinized using xylene for 5 to 10 minutes, followed by dehydration with 100% alcohol. The sections were then stained with hematoxylin for 3 to 4 minutes and counterstained with 0.5% eosin for 15 to 30 seconds to achieve a pale pink appearance. After blotting, the slides were cleaned with xylene for 15 to 30 seconds, mounted with DPX, and dried to remove any air bubbles.12<\/sup><\/p>\n Determination of Antioxidant parameters<\/strong><\/p>\n Antioxidants play a crucial role in cancer research and can be assessed through various parameters to understand their impact on cancer development, progression, and treatment. Antioxidants are compounds that protect cells from oxidative stress by neutralizing harmful free radicals, which can damage DNA and lead to cancer.13<\/sup><\/strong> Here are some key antioxidant parameters commonly studied in the context of cancer:<\/p>\n Superoxide Dismutase (SOD)<\/strong><\/p>\n One important enzyme in the body’s defense against oxidative stress is superoxide dismutase (SOD). It assists in transforming superoxide radicals into less dangerous compounds. With a final concentration of 0.1 M phosphate buffer (pH 6.5), 1 nM CDNB (1-chloro-2,4-dinitrobenzene) in 95% ethanol, and 1 mM GSH, the reaction volume (3 ml) was incubated for 5 minutes at 37 \u00b0C. The reaction was initiated by the addition of the enzyme sample, and for five minutes, the enzyme activity was monitored at 340 nm. 14<\/sup><\/p>\n Catalase (CAT)<\/strong><\/p>\n This antioxidant enzyme aids in the reduction of oxidative damage by dissolving hydrogen peroxide into oxygen and water. To start the reaction, 0.5 mL of hydrogen peroxide (H\u2082O\u2082) solution was added to 200 \u03bcL of diluted homogenate, 1.0 mL of phosphate buffer, and 0.4 mL of distilled water. In control test tubes, the hydrogen peroxide (H\u2082O\u2082) solution was not present. To halt the process, 2 mL of potassium dichromate acetic acid was added after it had been incubated for 1 minute at 37\u00b0C. After 15 minutes in a boiling water bath, the samples were cooled, and the absorbance at 570 nm was measured in comparison to the control.15<\/sup><\/p>\n Glutathione (GSH)<\/strong><\/p>\n Glutathione is an essential intracellular antioxidant that participates in detoxification processes and helps protect cells from oxidative damage.<\/p>\n Analytical statistics<\/strong><\/p>\n All of the data were compared using a one-way ANOVA and Dunnett’s posttest. Each outcome was reported as a rise or fall relative to the values of the control results. For all statistical studies, Graph Pad Prism version 8 was utilized. A variance was deemed statistically significant if p < 0.05.<\/p>\n Results<\/strong><\/p>\n Effect of plant extract on body weight<\/strong><\/p>\n As shown in table no 1 N-methyl-N-nitrosourea (NMU) was administered by intraperitoneal route, the body weight of the animals in the NMU control group significantly decreased (91.51\u00b10.45) from their starting weights. Simultaneous treatment of Tamoxifen (3.3 mg\/kg) and plant extracts of Gliricidia sepium<\/em> orally resulted in a considerable rise Standard group (122.3\u00b10.66), TEST 1 CHGS 200 mg\/kg (116.3\u00b11.64) and TEST 2 ETGS 200mg\/kg (119.5\u00b11.37) in body weight when compared to the initial weight of the animals.<\/p>\n Table\u00a0 1: Animal Body Wight among different treatment groups<\/strong><\/p>\n
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