Toxicology International

Modulatory Role of NTSR1 Antagonist SR48692 in HFD-Induced Thyroid Impairment in Mice: An In Vivo Study

Himanshu Pal — 2024-05-16

Thyroid hormones are one of the crucial regulators of metabolism, especially lipid, and is linked with lipid metabolism dysregulation. High-Fat Diet (HFD) induces the secretion of Neurotensin (NTS), which enhances lipid absorption, and deposition, proportionately increases leptin. Increased leptin is associated with central leptin resistance and affects thyroid regulation which may lead to hypothyroidism. We hypothesized that HFD-induced increased NTS may cause dyslipidemia leading to thyroid alterations, which could be antagonized by the NTS receptor-1 (NTSR1) antagonist SR48692. Swiss albino mice (n = 24) were divided into two sets and fed with Standard Diet (SD) and HFD for 4 weeks. After that each set of mice were further divided into two groups and administered with 0.9% saline and SR48692 (100 μg/kg bw) intraperitoneally for 4 weeks and maintained as Group-I: (Control/SD), Group-II: (HFD), Group-IV: (HFD+SR48692), and Group-IV: (SD+SR48692). HFD-fed mice showed significantly decreased T3 and T4 with increased TSH and leptin, suggesting leptin-induced hypothyroidism, also reflected in the histological alterations observed in thyroid and adipose tissue. Co-treatment with SR48692 (HFD+SR48692) showed amelioration in the thyroid, as levels of T3, T4, TSH, and resensitization of leptin, were evident in histopathological analysis compared to HFD suggesting the therapeutic efficacy of SR48692 on thyroid impairment in dyslipidemia. Only SR48692-treated group showed an exacerbating effect.

Poisoning Patterns, Causes, and Implications for Hospital-Centric Interventions- A Retrospective Single-Centre Observational Study from India

Jnanaprakash B. Karanth — 2024-05-29

Poisoning poses a significant public health concern in India, with rising trends observed over recent years. The study aims to observe patterns of demographic characteristics, presentation modes, outcomes, and associated factors among poisoning cases in India. A retrospective observational study was conducted on data of individuals admitted for poisoning from 2019 to 2023. Demographics, mode of presentation, type of poisoning, poisonous substances, presenting complaints, ICU admission, outcomes, time of ingestion, time of hospital admission, time delay in treatment initiation, and duration of hospital stay were analysed. Among the 357 individuals, the majority resided in rural areas and had up to a high school education. The majority of the admitted cases were self-poisoning (77.1%) followed by accidental poisoning (22.5%) and one homicide. Most of the self-poisoning cases (42.1%) were young adults (20-29 years), and accidental poisoning (32.9%) were individuals in their forties. Pesticides (67.9%), especially organophosphate and snake bites (100%) were the most common poisoning agents of self-poisoning and accidental poisoning, respectively. Mortality was 5.2% in self-poisoning, and 1.2% in accidental poisoning cases. Most patients who succumbed to the poisoning were in their thirties, with 78.6% requiring ICU admission. These events mostly occurred in autumn, during the daytime. Most patients were unconscious at admission. Self-poisoning using pesticides was the most common reason for admission. Seasons, poisoning agents, time of admission, presenting complaints and treatment delay contributed to the morbidity and mortality. Establishing poison information centres and toxicological units in hospitals is essential for improved management strategies.

DNA Hypomethylation could be a Predictive Biomarker for B Vitamin Deficiencies Among Working Population

Kalahasthi Ravi Babu — 2024-08-05

The methylation (CH₃) capacity is dependent on the availability of B vitamins in one carbon metabolism. This study evaluated B vitamin (B9 and B12) deficiency status and its impact on the Diagnostic Accuracy (DA) of DNA methylation among the working population. DNA methylation (5 methylcytosine) and B vitamins were quantified using the ELISA methods. Based on the cut-off values of B vitamins, the subjects were categorized into adequate, marginal, and deficiency status. Fagan’s nomogram was used to assess the DA of DNA methylation in different conditions of B vitamin. DA metrics are used in this study as a Diagnostic Odd Ratio (DOR), a positive Likelihood Ratio (LR+), a negative Likelihood Ratio (LR-), and a posttest probability of true positive DNA methylation that is less than 2.0%. DNA methylation was significantly lower in marginal and deficient B-vitamin status as compared to adequate status. The DA metrics at 5 mC showed high sensitivity and modest specificity for marginal and deficient B vitamins. In vitamin B9 deficiency, LR (+) was 2.51 (95% CI: 0.99–6.39), DOR was 10.4 (95% CI: 2.06–51.2), and the post-test probability was 92% (95% CI: 83–97). In vitamin B12 deficiency, LR (+) was 1.58 (95% CI: 1.03–2.44), DOR was 3.16 (95% CI: 1.29–7.66), and the post-test probability was 86% (95% CI: 80–90). In B vitamin deficiency, DA metrics of DNA methylation: LR+, LR-, DOR, and post-test probability were more precisely predicted. DNA hypomethylation could be used as a predictive biomarker in B vitamin deficiency. The findings of the study help to prevent adverse health effects due to methylation capacity loss.

The Effect of Tinospora cordifolia Leaf and Stem Aqueous Extracts on Zidovudine (ZDV) Induced Hematological Changes in Rat Model

Harinatha Reddy Aswarthagari — 2024-05-16

The present study aims to evaluate the protective role of aqueous extracts of stem and leaves of Tinospora cordifolia (AELTC and AESTC) on the hematologic toxicity effects of ZDV in Rats. A Soxhlet extractor was used to prepare separate aqueous extracts of stem and leaf. Twenty-four albino rats (weighing 200-230 g) were separated into four groups. Group I rats were administered distilled water (negative control). Group II rats were administered 500 mg/kg b.w of ZDV (positive control), Group III rats were administered 500 mg/kg b.w. of AELTC along with ZDV, and Group IV rats were administered 500 mg/kg b.w of AESTC along with ZDV. The hematological parameters of the rats were assayed during the 2nd, 4th, 6th, and 8th weeks of treatment. The animals exposed to Zidovudine showed a significant decrease in Red Blood Cells (RBC), Hemoglobin (Hb), White Blood Count (WBC), and platelet count compared to control (Group I). The results of T. cordifolia leaf and stem administered rats showed a significant elevation in RBC, Hb, WBC, and platelets count. Spleen histology showed immunological and hemorrhagic changes with an accumulation of RBCs and severe depletion of lymphocytes in white pulp were observed in ZDV-treated rats. The intake of AELTC and AESTC showed apparent changes in Spleen tissues. These results suggest that AELTC and AESTC stimulate the hematopoietic system, thereby increasing blood cell levels in ZDV-treated rats.

Heavy Metals in Fishes, Water and Macrophyte of the Ganga River and Risk Related to their Consumption

Sumit Kumar — 2024-05-30

The study was conducted along the Ganga River and aimed to assess the contamination levels of six heavy metals (Pb, Cd, Hg, Cu, Cr, and Zn) in fishes, water, and macrophyte (Eichornia crassipes) of Haridwar (Bhadrabad - A1), Bijnor (Near Ravidas temple-A2) and Muzaffarnagar (Bairaj Ganga bridge-A3) using a flame atomic absorption spectrophotometer (Avanta Σ). The findings revealed a concerning level of heavy metal contamination in various components of the Ganga River ecosystem. In fishes, the order of heavy metal occurrence was Cr>Zn>Pb>Cu>Cd>Hg, with Pb levels exceeding the recommended WHO/FAO limits in some samples. Water samples showed heavy metals in the order Pb>Cu>Zn>Cr>Cd>Hg, with elevated levels of Pb and Cr at site Haridwar (Bhadrabad - A1) exceeding WHO’s maximum permissible limits. Macrophyte analysis indicated heavy metals in the order Zn>Cr>Pb>Cu>Hg>Cd, with Pb and Cr levels surpassing WHO recommendations. The heavy metal bioaccumulation was higher during the post-monsoon and pre-monsoon seasons while lower in the monsoon season. Significant differences in heavy metal concentrations among different organs of fishes highlight the variability in metal accumulation and distribution. The study underscores the importance of regular biomonitoring to assess the risks posed by heavy metals, especially considering the significant reliance of the local population on the Ganga River for various purposes. Continued monitoring and implementation of mitigation measures are essential to safeguard human health and the integrity of the ecosystem in the face of heavy metal contamination.

Effect of Cymbopogon schoenanthus L. Spreng Extract on Gentamicin-Induced Renal Toxicity in Rats

Farah Ramdane — 2024-07-10

This study’s objective is to assess phytochemicals content and the protective effect of Cymbopogon schoenanthus hydroethanolic extract on gentamicin-induced renal toxicity in rats. Phytoconstituents were determined by using colorimetric tests. Three methods were used for the evaluation of antioxidant activity. The experiment used gentamicin in six different groups of albino rats. The biochemical analysis of current results displayed a significant reduction in blood urea and creatinine in groups intoxicated with gentamicin and administered two doses of the extract. While the concentration of total proteins and albumins has been increased. In addition, this plant decreased oxidative stress which was translated into a decrease in malondialdehyde content, and increased levels of antioxidant parameters, consequently an improvement in kidney functions. This study revealed a higher content of phenolic compounds, greater antioxidant activity and nephroprotective qualities of Cymbopogon shoenanthus extract for the first time.

GC-MS Profiling of Secondary Metabolites in Hexane and Ethyl Acetate Crude Extracts of Red Sea Soft Coral Xenia macrospiculata

Madhubabu Kasimala — 2024-08-05

Soft corals are macro-invertebrates found in marine ecosystems. The genus Xenia belongs to the family Xeniidae containing ninety-eight species, rich sources of bioactive natural products including terpenoids, steroids, and fatty acids. The hexane and ethyl acetate solvent extracts of soft coral Xenia macrospiculata Gohar, 1940 (X. macrospiculata) were investigated for the novel chemical constituents using Gas Chromatography-Mass Spectroscopy (GC-MS). Qualitative analysis of soft coral crude extracts has shown various compounds with high and low molecular weights. In the hexane extracts twentyfive compounds were identified including sesquiterpene derivatives (34.61%), steroid derivatives (30.76%), cyclic diterpenes (7.69%), oxygenated terpenes (7.69%), fatty esters (7.69%), mercaptans (3.84%), alcohols (3.84%), and bicyclic monoterpenes (3.84%). In ethyl acetate extract twenty compounds were identified, the major compounds are fatty acid derivatives (28.57%), element type sesquiterpenes (23.80%), steroid derivatives (19.04%), thio compounds (9.52%), alkaloids (4.76%), diterpenes (4.76%), alcohols (4.76%), Fluro-hydrocarbons (4.76%). GC-MS profiling of hexane and ethyl acetate extracts of X. macrospiculata revealed the presence of several potential bioactive compounds. The elemene, azulene, himachalene and aromadendrane sesquiterpenoids are unique compounds reported first in the soft coral X. macrospiculata. The available literature supports that these compounds have potential antioxidant, Tumour Necrosis Factor (TNF) inhibition and cytotoxic activity. Some compounds are used as tasteful natural sweeteners and some of the fatty ester derivatives are potential flavouring agents in food industries.

Influence of Processing Techniques on the Saponin Content and Nutritional Quality of Quinoa (Chenopodium quinoa)

Priyanka R. Kale — 2024-06-10

Quinoa cultivated primarily for its edible seeds is renowned for its nutritional richness. The study highlights how each method affects quinoa’s nutritional composition and saponin content by comparing different processing techniques. This information is valuable for food processors and manufacturers seeking to optimise their production processes to enhance their nutritional value. The soaking process involved immersing the seeds in sodium bicarbonate and citric acid solution (1% and 2%, respectively) for 6 hours at room temperature then roasting at 800C for 10 min. These pretreated quinoa grains were analyzed for their saponin content and nutritional composition. In raw quinoa grains, the saponin content was measured at 0.64%. After soaking the grains in a 2% citric acid solution, a significant reduction in saponin content to 0.09%. However, roasting the quinoa grains results in a decreased saponin content slightly to 0.62%. The results indicate that soaking in a 2% citric acid solution was particularly effective in reducing the saponin content of quinoa seeds to a desirable level while also eliminating bitterness. Consequently, quinoa seeds treated with a 2% citric acid solution were selected for further nutritional analysis. Significant alterations in the nutritional composition of quinoa were observed as a result of processing. In terms of proximate composition, soaking increased moisture, protein, and ash while decreasing other parameters. Soaking causes an increase in protein content from 15.6% to 16.2%. Conversely, roasting decreased moisture and protein levels while increasing other parameters. The moisture and protein content of roasted quinoa grains were reduced to 6.8% and 14.8%, respectively. Both soaking and roasting processes resulted in elevated mineral content in quinoa seeds. Consequently, soaked and roasted quinoa grains were found to be rich in nutrients, positioning them as valuable sources for enhancing food commercialization.

Azoxystrobin Induced Changes in the Gill Histoarchitecture, Brain Acetylcholinesterase Activity and the Behavior of the Fish Pethia conchonius from the River Teesta

Arpita Ray — 2024-07-10

Azoxystrobin is a globally used strobilurin fungicide, which contaminates waterbodies through surface run-off. Its bioaccumulation in aquatic animals via food chains can induce serious pathophysiological disturbances. Therefore, histopathological and neuronal effects of azoxystrobin have been assessed in the fish, Pethia conchonius in the laboratory condition. Azoxystrobin-treated fish showed slow movement, crowding at the bottom, loss of equilibrium, and excess mucus secretion at all concentrations (0.025mg/L, 0.0514 mg/L, and 0.103mg/L) at 48 hours of exposure compared to the control. A significant dose and time-dependent inhibition in acetylcholinesterase activity was observed (p<0.05). The highest acetylcholinesterase inhibition (45.45 ± 1.07) was noted for the highest concentration at 96 hours of exposure than the control groups (88.35 ± 0.71). In contrast to the control, different histopathological changes in gill tissues have been observed like, epithelial lifting, lamellar fusion, epithelial hyperplasia, and the curling of secondary lamellae in the azoxystrobin-exposed groups after 24 hours of treatment. The results of this study indicated that azoxystrobin is neurotoxic as well as damaging to gills.

Effect of Iron Overload on Tight Junctions and Adhesion Molecules in ECV304 Cells

Samed Refik Sar — 2024-06-10

Blood vessels are essential for the body’s tissues and organs to receive oxygen and nutrition. The Blood-Brain Barrier (BBB) is a special feature of the blood arteries that vascularise the Central Nervous System (CNS) which enables these vessels to tightly control the flow of ions and molecules between the blood and the brain. The accurate regulation of CNS homeostasis facilitates appropriate neuronal performance and safeguards neural tissue from toxins and pathogens. Modifications to these barrier characteristics play a significant role in the development of many neurological disorders. BBB contains tight junction transmembrane proteins, integral membrane proteins, occludin, claudins, IgG-type proteins, junction adhesion molecules and scaffold proteins. Numerous biological functions require iron in the CNS, including neurotransmitter synthesis, myelin formation and mitochondrial function. However, excess iron can lead to oxidative stress and damage, which are implicated in neurodegenerative diseases. In this research, the impacts of iron accumulation on Cadherin 5 (CDH5), Claudin 5 (CLDN5), Intercellular Adhesion Molecule 1 (ICAM-1), Occludin (OCL), p-selectin (P-SEL), Vascular Cell Adhesion Molecule 1 (VCAM-1) and Zonula occludens-1 (ZO-1) genes expressions in Human Umbilical Vein Endothelial Cells (ECV 304) cells were investigated. It was found that in human umbilical vein endothelial cell line cells, iron overload enhanced the expression of CDH5 and P-SEL genes while reducing the expression of VCAM1, Cldn5, ICAM-1, OCL, and Zo-1 genes.

Ameliorative Effect of SR48692 on the Testicular and Epididymal Impairments of High-Fat Diet- Induced Swiss Albino Mice

Pradeep Verma — 2024-07-24

This study investigated the role of Neurotensin (NT) receptor-1 antagonist SR48692 on High-Fat Diet (HFD)-induced considerable morphological alterations in the testis and epididymis in mice. Excessive intake of fat and a sedentary lifestyle are major causes of metabolic comorbidities in the current obesogenic environment. Consumption of HFD induces NT release in the gut, which promotes lipid absorption. Elevated NT is associated with metabolic alterations and imbalanced energy status which could be responsible for a negative impact on reproductive outcomes. Male mice (n = 24) were randomly assigned, based on diet into Control Diet (CD) and HFD for 4 weeks; they were further divided into 2 subgroups and administered saline and SR48692 (100 μg/kg b.w.) intraperitoneally for the next 4 weeks. The present findings revealed that HFD-induced obesity caused significant alterations in circulating lipid and reproductive hormone profiles leading to deterioration of testicular and epididymal histoarchitechture. However, the co-treatment of SR48692 with HFD has significantly restored the HFD-induced detrimental effect on lipid and reproductive hormone profiles, justified by the improvements in histoarchitechture of the testis and epididymis. However, only SR48692 administered group showed adverse effects on the studied parameters. The results reported herein showed that SR48692 have the efficacy to actively overcome the HFD-induced detrimental effects. Therefore, SR48692 might be considered for the therapeutic role in HFDinduced reproductive impairments.

Therapeutic Potential of Alcoholic Extract from Lemna minor Linn. in Attenuating Atherosclerosis Induced by High-Fat Diet in Male Wistar Rats

B. R. Yamini — 2024-07-24

This study explored the cardioprotective effects of Alcoholic Extract of Lemna minor Linn. (AELM) against high-fat diet-induced atherosclerosis in male Wistar rats. AELM was administered at doses of 300 mg/kg and 800 mg/kg. Assessments included lipid profiles, oxidative stress markers, cardiac injury enzymes, and liver parameters associated with cholesterol synthesis. Results showed significant reductions in lipid levels, oxidative stress markers, and cardiac injury enzymes, particularly with the higher dose of 800 mg/kg. AELM also lowered the atherogenic index and improved HDLc levels, indicating the potential to mitigate atherosclerosis-related lipid imbalances. Furthermore, AELM exhibited a dose-dependent reduction in the HMG CoA/mevalonate ratio, suggesting inhibition of HMG-CoA reductase activity, crucial in cholesterol synthesis. Molecular docking studies supported AELM’s antiatherosclerotic potential, with leucine demonstrating favourable binding energies with atherosclerosis-associated enzymes. Histopathological analysis revealed structural improvements in rat aortas with AELM treatment. In conclusion, AELM presents promise as a therapeutic agent against atherosclerosis through modulation of lipid metabolism, attenuation of oxidative stress, and inhibition of HMGCoA reductase activity.

Biomonitoring of Serum Markers as a Measurement of Occupational Exposure in Personnel at Landfill Site

Diya Fulwani — 2024-08-05

Improper handling and management of municipal solid wastes can produce adverse health effects on waste pickers through biological, chemical, physical, mechanical or psychological means. They are vulnerable to various occupational health hazards such as respiratory disorders, gastrointestinal diseases, skin diseases, eye infections and musculoskeletal disorders. Hence, we aimed to conduct biomonitoring and health risk assessment on women rag pickers. This study included 250 women waste pickers: 160 exposed working on the Pirana dumping site and 90 control subjects from around the residential areas of Ahmedabad city of Gujarat (India). C-reactive protein, Alkaline phosphatase, Alanine transaminase and Aspartate transaminase were significantly high in serum samples of exposed subjects suggesting gallbladder inflammation, acute hepatitis A and B or oxidative stress. After comparing the lipid profile of exposed and control subjects, we found a significant decrease in cholesterol, high-density lipoprotein cholesterol and triglyceride frequency which is a sign of malnutrition or hyperthyroidism. Random blood sugar, globulin and total serum protein were reduced significantly. In contrast, a significant increase was found in albumin, phosphorus, creatinine, urea and uric acid levels of exposed individuals, which indicates potential disorders of the liver and kidney. Direct exposure to solid waste, agrochemicals, heavy metals, toxic industrial and hospital waste, and toxic air having high concentrations of polyaromatic hydrocarbons and bioaerosols, unhealthy eating and sanitary habits, and negligence towards health were major contributing factors to ill effects on their liver and kidney. We suggest effective education, and communication programs for preventive and protective measures to address problems of rag pickers. The present study emphasizes the need for regular monitoring of contamination among the rag-picking population and areas around the Pirana landfill site for better information on exposure to rag pickers.

In Vitro Evaluation of Cytotoxicity Induced by Indoxacarb and its Amelioration with Cassia fistula Bark Extract

Zarzoliani — 2024-08-05

The primary objective of the study is to evaluate the cytotoxic effect and biochemical alteration on exposure to the Indoxacarb (IDC) in the Madin-Darby Bovine Kidney (MDBK) cell line and its amelioration by Ethanolic Bark Extract of Cassia fistula (EBECF). The MTT assay was used to study the cell viability and determine the IC₅₀ value. The cells were further treated with IC₂₅, IC_12.5 and IC_6.25 values of IDC alone and in combination with EBECF for 24, 48 and 72 hours. IDC-induced cytotoxicity was evident by cellular morphological changes such as rounding of cells, cellular swelling, blebs formation, nuclear degradation, intracytoplasmic changes and loss of intracytoplasmic contents. The biochemical alteration was observed as a significant increase (p<0.05) in the levels of Lipid Peroxidation (LPO) and cytotoxicity marker enzymes namely Lactate Dehydrogenase (LDH), Alkaline Phosphatase (ALKP), Acid Phosphatase (ACP) and Creatine Kinase (CK). These changes were dose-dependent in IDC-treated groups. EBECF intervention in IDC-treated cells ameliorated the toxicity against lower concentrations. Thus, the present study suggested that C. fistula bark has the potential to protect the cells from IDC-mediated toxicity to a limited extent.

Evaluation of Respiratory Toxicity Potential of Acrylamide in Zebrafish: Oxidative Stress Parameters, mRNAs Expression, and Histomorphological Changes in Gills

Harsh R. Patel — 2024-08-05

Anthropogenic contamination of surface water by chemicals is a worldwide concern, which affects human health and aquatic organisms including fish. Acrylamide (ACR), a Neo-Formed Contaminants (NFC) leads to serious toxic effects on the respiratory system of marine inhabitants. The mechanisms underlying ACR-induced oxidative stress and altered mRNA expressions in zebrafish are unclear. The present investigation aimed to evaluate the effects of ACR exposure for 28 days on the activity or level of antioxidant response elements (ARE), mRNA expressions of antioxidant genes and histomorphological changes in the gills of adult zebrafish. The 270 adult zebrafish were randomly allocated into 03 experimental groups viz. control group, T1 group (ACR: 8.5 mg/L) and T2 group (ACR: 17 mg/L). After 28 days of ACR exposure, Superoxide Dismutase (SOD) activity was significantly (p<0.01) decreased in the gills of zebrafish of the T2 group; and no change in Catalase (CAT) activity was observed in both the treatment groups. The levels of reduced Glutathione (GSH) were significantly (p<0.001) decreased and Malondialdehyde (MDA) were significantly (p<0.001) increased in a concentration-dependent manner. The sod and nuclear factor erythroid 2-related factor 2 (nrf2) mRNA expressions were significantly (p<0.001) downregulated in the gills of zebrafish of both treatment groups. Noticeable histomorphological alterations were recorded in the gills of zebrafish of the T2 group. Alterations in ARE activity, mRNA expressions of antioxidant genes and histopathological findings suggest ACR exposure has been shown to produce oxidative stress-mediated damage in the gills of adult zebrafish.

Therapeutic Potential of Curcuma longa Rhizome Solvent Fractions Against Benz[a]Anthracene-induced Pulmonary Toxicity: In vivo Insights

Oladipo Miriam Oluchi — 2024-08-05

Benz[a]anthracene, a polycyclic aromatic hydrocarbon found in environmental pollutants, poses significant health risks, particularly to the respiratory system. This study aimed to evaluate the potential therapeutic effects of different solvent fractions of Curcuma longa rhizome methanol extracts against benz[a]anthracene-induced pulmonary toxicity using an in vivo rat model. The methanol crude extract of C. longa was partially fractionated using partition chromatography with silica gel as the adsorbent. Four solvent fractions obtained using gradient elution (designated ethanol, ethyl acetate, diethyl ether, and n-hexane fractions) were collected and used in the experiment. Albino Wistar rats (Rattus norvegicus) were randomly divided into seven groups and administered benz[a]anthracene to induce pulmonary toxicity. Subsequently, rats were treated with different solvent fractions of C. longa or diclofenac sodium as a positive control. Body weights were measured weekly, pre-induction, post-induction and post-treatment blood samples were collected, and lung tissue samples were collected and analysed. Treatment with the ethyl acetate fraction of C. longa resulted in significant improvements in lung histopathology compared to benz[a]anthracene-treated rats alone, suggesting potential anti-inflammatory effects. Furthermore, this fraction showed significant antioxidant enzyme activities, including glutathione peroxidase, glutathione reductase, and superoxide dismutase, indicating its ability to mitigate oxidative stress. Biochemical analysis revealed that the ethyl acetate fraction also significantly decreased serum levels of tumour biomarkers, such as epidermal growth factor receptor, cancer antigen 125, and carcinoembryonic antigen, suggesting its potential in inhibiting cancer-related processes induced by benz[a]anthracene exposure. Our findings suggest that the ethyl acetate fraction of C. longa rhizome may hold promise as a therapeutic agent against benz[a]anthracene-induced pulmonary toxicity, possibly through its anti-inflammatory, antioxidant, and anticancer properties. Further research is warranted to elucidate the specific compounds present in this fraction and the underlying mechanisms through which they exert their effects, to optimize their therapeutic potential.