GST Activity Research Articles

Development of new LSM-83177 analogues as anti-tumor agents against colorectal cancer targeting p53-MDM2 interaction

LSM-83177, a phenoxy acetic acid derivative, is a small molecule reported for its promising anti-tumor properties. Via inhibiting the interaction between MDM2 and p53, LSM-83177 can elevate the active p53 levels within cells, thereby promoting apoptosis and inhibiting tumor growth. Also, LSM-83177 has been shown to inhibit GST activity in colorectal cancer HT29 cells. In the current work, novel LSM-83177 hydrazone analogs 5a-f, 7a-b, 10a-e, and 13a-b have been designed according to the structure features of LSM-83177 and their binding mode in the active site of MDM2. The anti-cancer activity of the newly synthesized analogs is evaluated against the HT29 cell line. The most potent compounds, 7a and 10a, showed IC50 = 12.48 and 10.44 µg/ml, respectively, when compared with Cisplatin (IC50 = 11.32 µg/ml) as a reference drug. Compounds 7a and 10a were introduced for further inspection for p53-MDM2 protein–protein interaction, where they displayed IC50 values of 3.65 and 11.08 µg/ml, respectively. Furthermore, hydrazones 7a and 10a increased the p-53 expression levels by 3.22– and 4.25-fold, respectively; in addition, they effectively reduced the GST expression levels in HT29 cancer cells with 0.56- and 0.30-fold increments in comparison to the untreated control.

Establishment of a cell culture from Daphnia magna as an in vitro model for (eco)toxicology assays: Case study using Bisphenol A as a representative cytotoxic and endocrine disrupting chemical

Bisphenol A (BPA) is a widely used industrial compound found in polycarbonate plastics, epoxy resin, and various polymer materials, leading to its ubiquitous presence in the environment. The toxicity of BPA to aquatic organisms has been well documented following in vivo exposure scenarios, with known cytotoxic and endocrine-disrupting effects. As such, BPA was used in this study as a well-characterized chemical to implement more ethical and resource-efficient scientific practices in toxicity testing through new approach methods (NAMs). Due to the frequent use of Daphnia spp. as a model organism in toxicology, we developed an in vitro cell culture system from Daphnia magna embryos, with optimized medium to support cell longevity. The cultures were maintained for up to two months, demonstrating their stability and suitability for cytotoxicity studies. Using this novel system, lethal concentration 50 (LC50) values were determined at the 24 and 48 h time points following BPA exposure. Subsequently, oxidative stress, endocrine disruption, and DNA damage were assessed through gene expression, activity assays, and a comet assay in BPA-exposed cells. LC50 values of 52 µM and 20 µM BPA were calculated after 24 and 48 h exposures, respectively. BPA cells exposed to 20 and 52 µM had significantly increased GSH, GPx, and GST activity levels. mRNA expression analysis revealed significant upregulations in the mRNA expression of hsp70, hsp90, gst, gpx, vtg1, and cyp4, with downregulations of sod, cat, and ecr following BPA exposure. Furthermore, comet assays showed a significantly higher level of DNA damage induced by BPA compared to controls, with greater comet and tail lengths. This study established a novel in vitro Daphnia model, using BPA as a case study for determining toxic effects, further highlighting the importance and applicability of utilizing alternative methods in ecotoxicological research through reducing animal use.

Genotoxic impact of agricultural insecticides as contaminants of river Teesta on the resident fish Pethia Conchonius.

Fish, being highly sensitive to changes in the physico-chemical parameters of water, are good indicators of contamination. Teesta, a prominent northern West Bengal River system, is increasingly contaminated due to anthropogenic activities. This study aims to determine agricultural pesticide contamination and its genotoxic impact on the resident fish, Pethia conchonius, as an experimental organism. Sample water analysis from three riverine sites I, II & III, showed the presence of the insecticides imidacloprid (IMI), chlorpyrifos (CPF), bifenethrin (BF), cypermethrin (CP), difenthiuron, acetamiprid (AC) in the sites II and III only with adjoining agricultural lands. Comet assay revealed a significantly lower % Head DNA (~ 1.2 times), higher %Tail DNA (~ 16 times), and %Tail length (~ 3.1 times) in the gills of Pethia conchonius from sites II and III. About 4 and 10 times increase of micronuclei and other nuclear abnormalities were also noted in the erythrocytes of the fish from sites II and III than I, which was not contaminated. The antioxidant enzymes SOD, CAT, and GST activity and MDA levels were significantly higher (p < 0.05) in the liver samples from sites II and III, while AChE activity was significantly decreased (p < 0.001) in the brain tissues. Moreover, the sod, cat, and gpx expression in the hepatic cells were significantly upregulated compared to the β actin mRNA indicating increased oxidative stress. Increased genomic damage, antioxidant enzyme activity, higher MDA levels, decreased AChE activity in the brain, and the upregulation of hepatic genes strongly suggested the genotoxic effects of the detected insecticides in combination with other contaminants.

Toxicity of polystyrene nanoplastic and copper oxide nanoparticle in Artemiasalina: Single and combined effects on stress responses

Nanoparticles, such as copper oxide nanoparticle (CuO NP) and polystyrene nanoplastic (PSNP), are increasingly released into aquatic environments, and pose potential risks to aquatic animals such as brine shrimps. Understanding the toxicity of these nanoparticles, especially when combined, is very important to assess their environmental effects. Therefore, this work describes the toxicity of polystyrene nanoplastic (PSNP) and CuO nanoparticles (CuO NPs) for brine shrimp (Artemiasalina). The body length and stress biomarkers, including the activity of SOD, CAT, GST, Acid phosphatase, AChE, level of MDA and GSH, and expression of the hsp70 gene were quantified. The 48h-EC50 values for PSNP, CuO NPs, and their combination were determined as 1.024 and 5.089, and 0.512 mg L−1, respectively. The combined exposure groups showed the highest growth inhibition. This was associated with increased activity of SOD and GST, decreased activity of CAT, a significant decrease in the level of GSH, a significant increase in the MDA level, and expression of the hsp70 gene (P < 0.05). Moreover, an increased ACP and reduced AChE activity were observed in exposure groups. This study indicated that PSNP and CuO NPs have synergistic toxicity for A.salina, underscoring the importance of further investigation into their combined effect on aquatic animals.

Copaiba Oleoresin Improves Weight Gain and IL-10 Concentration, with No Impact on Hepatic Histology, in Liver Cirrhosis.

Copaifera sp. is a native tree in the Amazon region. Copaiba oleoresin has components such as sesquiterpenes, which have anti-inflammatory and antioxidant potential. Liver cirrhosis is the end stage of liver disease with limited therapeutic options. We aimed to evaluate the effect of copaiba oleoresin supplementation on the liver of animals with thioacetamide (TAA)-induced cirrhosis. For the induction of liver cirrhosis, 100 mg/kg of TAA was administered intraperitoneally twice a week for 8 weeks. A total of 200 mg/kg/day of copaiba oleoresin was administered via gavage for the same period. Copaiba oleoresin supplementation improved cirrhosis-associated cachexia by increasing weight gain and body fat. In addition, copaiba oleoresin attenuated systemic inflammation, as shown by the decrease in the circulating C-reactive protein. In the liver, the copaiba oleoresin decreased carbonyl proteins and increased IL-10 compared with TAA-treated rats. TAA groups demonstrated increased SOD, catalase, GST, and GSH activity in the liver. In conclusion, the supplementation of copaiba oleoresin demonstrated a beneficial systemic effect in alleviating cirrhotic cachexia and antioxidant and anti-inflammatory action in the liver. However, it failed to improve the serological and histological markers of liver damage, which could be associated with the advanced stage of the disease.

Evaluating Antioxidant Enzyme Parameters in Rats Treated with Berberine-Loaded Bovine Serum Albumin Nanoparticles and Scopolamine

Abstract Introduction/Objective Introduction: Berberine (BER) has garnered attention for its antioxidant properties demonstrated in various studies, showing its ability to scavenge or inhibit reactive oxygen species (ROS) and reactive nitrogen species (RNS), including nitric oxide, superoxide anion, and peroxynitrites. Methods/Case Report Methods: Forty-two male Wistar rats were randomly assigned to seven groups, each comprising 6 rats: Sham control (no treatment), BSA-NPs orally administered, BRB-NPs orally administered, SCO intraperitoneally injected, BER followed by SCO injection, BSA-NPs with SCO injection, and BER-NPs with SCO injection. Treatments were administered daily for 28 days. The cholinergic status was evaluated by assessing acetylcholinesterase (AChE) activity in brain homogenates, and brain homogenate Aβ-42 levels were measured using the ELISA technique. Additionally, brain prooxidant markers (NO, TBARS, XO), nonenzymatic antioxidants (GSH), and enzymatic antioxidants (GPx, GST, and SOD) were measured. Results (if a Case Study enter NA) Results: The study revealed that treatment with BRB NPs resulted in the lowest AChE activity and Aβ-42 levels. Scopolamine administration significantly increased brain NO, TBARS, and XO activity, accompanied by a progressive depletion in GSH content (p&amp;lt;0.05). Treatment with BRB-free or BSA decreased NO levels, while the BSA or BRB-NPs groups exhibited higher brain TBARS levels than the sham group. All treated groups exhibited similar activity to the sham control, except for the BSA-treated group. Furthermore, all treated groups showed comparable GSH levels to the sham control, except for the scopolamine group. Treatment with BRB NPs normalized GPx activity and showed the highest GST activity. The BRB-NPs treated group also exhibited the highest SOD level. The p-value was &amp;lt;0.001 among all studied groups. Conclusion Conclusion: Injection of SCO induced oxidative impairments in brain tissue, evidenced by increased prooxidants such as NO and depletion of antioxidant capacity (GPx, GST, and GSH). SCO also disrupted glutathione metabolism, enhancing the level of lipid peroxidation in the brain. Treatment with BRB-NPs or free BRB progressively increased antioxidant capacity and mitigated the harmful effects of oxidative damage in the brain by reducing prooxidant levels.

Reno-protective potential of poncirin against polyethylene microplastics instigated kidney damage in rats via regulating Nrf-2/Keap-1 pathway

Polyethylene microplastics (PEMPs) are noxious environmental pollutants that are documented to cause organ damage including the kidneys. Poncirin (PON) is a naturally occurring flavonoid which demonstrates a wide range of pharmacological properties. This experiment was conducted to evaluate the palliative potential of PON against PEMPs induced renal toxicity by examining a range of biochemical and physiological parameters. Twenty-four male albino rats were randomly apportioned into four distinct groups including the control, PEMPs (1.5 mgkg−1), PEMPs (1.5 mgkg−1) + PON (20 mgkg−1) and only PON (20 mgkg−1). Our results displayed that PEMPs intoxication escalated the levels of urea, KIM-1, creatinine and NGAL while reducing the creatinine clearance level. Besides reduction in the activities of GPx, GST, HO-1, CAT, GSR & upsurge in the levels of MDA and ROS were detected in PEMPs group. Conversely, the levels of inflammatory markers including COX-2, IL-6, IL-1β, NF-kB and TNF-α were augmented following the PEMPs intoxication. Besides, the results of the current research demonstrated that the expressions of Bax and caspase-3 were esclated whereas the Bcl-2 expression was lowered from its standard value due to PEMPs provision. However, PON treatment significantly restored the PEMPs-induced aforementioned impairments. Therefore, PON could be used as a therapeutic compound to ameliorate PEMPs-induced kidney impairments in rats, possibly due to its tremendous pharmacotherapeutic potential.

Hesperetin-7-rutinoside mitigates erythrocyte dyslipidemia and oxidative stress induced by dichlorvos

In many rural and semi-urban areas of Nigeria, dichlorvos (DDVP) is a commonly utilized pesticide. However, it provokes remarkable health hazards, including erythrocyte toxicity. Hesperetin-7-rutinoside (Hesp-7-Rut), a citrus flavanone glycoside with antioxidant, anti-dyslipidemia and acetylcholinesterase-modulating properties has been investigated for its protective effects against various xenobiotic-mediated toxicities. This inquiry probed the chemotherapeutic role of Hesp-7-Rut in mitigating erythrocyte damage in rats. Forty-two rats were randomly `apportioned into seven groups (6 rats/group): control, DDVP alone (8 mg.kg⁻¹day⁻¹), DDVP + Hesp-7-Rut (50 mg.kg⁻¹day⁻¹), DDVP + Hesp-7-Rut (100 mg.kg⁻¹day⁻¹), DDVP + atropine (0.2 mg.kg⁻¹day⁻¹), Hesp-7-Rut only (50 mg.kg⁻¹day⁻¹), and Hesp-7-Rut only (100 mg.kg⁻¹day⁻¹). Rats were orally administered DDVP for 7 days followed by Hesp-7-Rut or atropine for another 14 days. Blood samples were collected to assess for biochemical assays. Hesp-7-Rut significantly (p < 0.05) rescinded DDVP-prompted increases in erythrocyte nitric oxide, malondialdehyde, cholesterol, phospholipids, and cholesterol: phospholipids ratio. Additionally, Hesp-7-Rut reversed DDVP-elicited decreases in red blood cell glutathione levels, activities of GST, SOD, catalase, glutathione peroxidase, and acetylcholinesterase. Overall, Hesp-7-Rut efficiently counteracts DDVP-elicited erythrocyte dysfunction by mitigating oxidative stress, dyslipidemia, and acetylcholinesterase inhibition. These findings highlight the potential of Hesp-7-Rut as a promising therapeutic agent for mitigating the harmful effects of DDVP exposure.

Chemical analysis of Alliin-Rich Allium sativum (Garlic) extract and its safety evaluation in Drosophila melanogaster

Garlic (Allium sativum) has been traditionally valued for its medicinal properties attributed to the presence of organosulfur compounds. Despite its benefits, concerns about herbal extract toxicity have arisen, necessitating safety assessment. This study was designed to evaluate the chemical analysis and safety profile of Alliin-Rich Garlic Extract (ARGE) using Drosophila melanogaster as a model organism. The ARGE was extracted from garlic cloves (Allium sativum Linn: UIH-23262) using a microwave-assisted method and characterized using UPLC-ESI-MS, 1H NMR, HPLC and IR. Its safety evaluation was determined using D. melanogaster (Harwich strain), and various assays were conducted on 1–3-day-old flies. Toxicological markers and oxidative stress were assessed to understand the impact of ARGE on the flies. Chemical profiling of ARGE using UPLC-ESI-MS, confirmed the presence of alliin (S-ally-L-cysteine-S-oxide), L-arginine, γ-glutamylmethionine, S-(2-carboxypropyl) glutathione, N-γ-glutamyl-S-(1-propenyl) cysteine, N-γ-glutamyl-S-(2-propenyl) cysteine, N-γ-glutamylphenylalanine, S-(allylthio) cysteine, γ-glutamyl-S-allylthiocysteine and eruboside B. HPLC confirmed an alliin content of 0.073 mg/g. Toxicological assessment in D. melanogaster revealed that ARGE enhanced antioxidant defenses by increasing total thiol levels and GST activity, while reducing acetylcholinesterase activity. No significant alteration was observed in catalase activity and cellular metabolic rate. Histological examination revealed no alterations in the histoarchitecture of the brain, fat body or gut of D. melanogaster. The study demonstrated the safety of ARGE in D. melanogaster, supporting its potential as a safe herbal remedy.

Myricetin inhibits CYP3A4, GST, and MRP1 in hepatic cancer cells

AbstractHerbal and nutritional supplements are widely used to prevent and treat many diseases, including cancer. Tumor cells modify metabolic enzyme systems like CYP3A4 and GST. They also overexpress MRP1, an ATP-binding cassette transporter subfamily G (ABCG2) member. Drug efflux may increase, reducing tumor cell drug accumulation and developing drug resistance that leads to significant obstacles in cancer care. Natural products' ability to overcome cancer's multidrug resistance is interesting. Their ability to affect several targets makes them valuable in addressing drug resistance from diverse approaches. The potential of natural flavonoid; Myricetin (MYR) to modulate CYP3A4, GST, and MRP1 activity and expression in hepatic cancer cells was evaluated to prove its targeting and preventing these pathways of multidrug resistance. The cell proliferation of MYR was determined using an MTT assay. Specific enzyme assays, efflux assay, and gene expression using RT-PCR were used to evaluate MYR effect in hepatic cell lines HepG-2 and Huh-7. MYR has a noteworthy cytotoxic effect compared to doxorubicin (DOX) with IC50 &gt; 100 μM in HepG-2 and Huh-7 cells. MYR showed potent inhibition of CYP3A4 and GST enzyme activity and MRP1 efflux function and downregulated their gene expression in a dose-dependent manner in both cells. MYR100 dose was the most significant effective dose. MRY100 decreased CYP3A4 activity by 67.5% (P &lt; 0.05) and 55% (P &lt; 0.01) and downregulated the gene by 0.2-fold (P &lt; 0.001) and 0.3-fold (P &lt; 0.001) in HepG-2 and Hub-7 cells, respectively. After treatment with MRY100, GST activity decreased significantly in both cells, reaching 47.6% (P &lt; 0.001) and 33.2% and GST gene downregulation was 0.12 and 0.21-fold (P &lt; 0.001). MRY100 inhibited MRP1 efflux pump 2.3 times (P &lt; 0.001) and 1.9 times (P &lt; 0.001) more effectively than PC, resulting in a 0.23-fold and 0.12-fold downregulation of MRP1 genes in HepG-2 and Hub-7 cells. The result will validate the use of MYR to interact with the metabolism phases and could be used as adjuvant therapy in cancer prevention and treatment approaches.

Assessment of Neurotoxic Mechanisms of Individual and Binary Mixtures of Cobalt, Nickel and Lead in Hippocampal Neuronal Cells.

Many studies have focused on the neurotoxic effects of single metals, while investigation on the exposure to metal mixtures, which mainly occur in real-life situations, is scarce. This study sought to assess the neurotoxic effect of Ni, Co, and Pb binary mixtures and their individual effects in hippocampal neuronal cells (HT-22). Cells were exposed to Ni, Co, and Pb separately for 48 h at 37°C and 5% CO2, and cell viability was assessed. Morphological assessment of the cells exposed to binary mixtures of Co, Ni, and Pb and single metals was assessed using a microscope. Furthermore, acetylcholinesterase (AChE) activity, oxidative stress biomarkers (glutathione [GSH] and malondialdehyde [MDA] levels, catalase [CAT], and glutathione-S transferase [GST] activities) and nitric oxide [NO] levels were evaluated after treatment with the binary mixtures and single metals. Binary mixtures of the metals reduced cell viability, exerting an additivity action. The combinations also exerted synergistic action, as revealed by the combination index. Furthermore, a significant reduction in AChE activity, GSH levels, CAT and GST activities, and high MDA and NO levels were observed in neuronal cells. The additive interactions and synergistic actions of the binary mixtures might contribute to the significant reduction of AChE activity, GSH levels, GST, and CAT activities, and an increase in MDA and NO levels. The findings from this study revealed significant evidence that binary mixtures of Co, Pb, and Ni may induce impaired neuronal function and, ultimately, neurodegeneration.

The effects of Selenium phytotoxicity on two wheat (Triticum aestivum) cultivars differing in Se tolerance and the role of antioxidant enzymes in the tolerance mechanism

Abstract Wheat seedlings were hydroponically grown in Hoagland solution containing various levels of Se. Tolerance response to Se toxicity was investigated by determining the level of thiobarbituric acid reactive substance (TBARS), proline and chlorophyll content, the growth parameters, and the activity of antioxidant enzymes. The toxic level of Se treatment significantly retarded the seedling growth. A substantial amount of proline accumulation was also observed in response to toxic Se concentration, but it was more pronounced in putative-sensitive cultivars. Chlorophyll content significantly decreased in Se-intoxicated seedlings and increased at the lowest Se dose in both cultivars. Severe and mild chlorosis was observed in putative-sensitive and tolerant cultivars at the highest Se level. Alterations in the activities of glutathione reductase (GR, 1.6.4.2), glutathione S transferase (GST, 2.5.1.18), guaiacol peroxidase (GPX, 1.11.1.7), catalase (CAT, 1.11.1.6), and ascorbate peroxidase (APX, 1.11.1.11) and superoxide dismutase (SOD, EC 1.15.1.1) were determined. TBAR level did not significantly increase in putative tolerant cultivars as an indicator of membrane lipid peroxidation. However, a significant increase was observed in putative-sensitive cultivars in response to higher selenium concentrations. In higher Se treatment groups, CAT and GST activities significantly increased in putative Se tolerant cultivars. However, excluding SOD, the activity of all the studied enzymes was increased considerably in putative-sensitive cultivars in a dose-dependent manner. Higher antioxidant enzyme activities and a substantial amount of proline accumulation did not significantly contribute to overcoming Se phytotoxicity in wheat seedlings grown in media supplemented with toxic selenium levels.

Rho4 interacts with BbGDI and is essential for the biocontrol potential of Beauveria bassiana by maintaining intracellular redox homeostasis

Rho4 is a member of the Rho-family small GTPases. In this study, we revealed the function of Rho4 and explored its mechanism involved in intracellular redox homeostasis in Beauveria bassiana, one of the most widely utilized filamentous entomopathogenic fungi. The disruption of Rho4 in B. bassiana resulted in significant phenotypic changes, such as fungal virulence, growth rate on different media, thermotolerance, germination, and conidiation. Integrated analysis of proteomic and transcriptomic data unveiled differential expression patterns of various redox-related genes and proteins in Δrho4, including the down-regulation of GST shown in proteomic and transcriptomic data, and the down-regulated gene expression levels of NOX, SOD, CAT, and GR in the transcriptome. Based on the bi-omics analysis, we focused on the impact of Rho4 in maintaining intracellular redox homeostasis. A decreased ROS content observed in Δrho4 might be attributed to the reduced NOX activity, which subsequently affects the GSH-producing/consuming metabolisms, with the attenuated activities of GR and GST. The imbalanced redox homeostasis also resulted in the reduced enzyme activities of SOD and CAT. Exogenous oxides could partially complement the ROS level and rescue the growth defect in Δrho4 to a certain extent. Besides, BbGDI was initially identified as an interacting protein of Rho4 in entomopathogenic fungi. Our results provide a comprehensive understanding of the function and regulating mechanism of Rho4 in B. bassiana.

Green synthesis of silver nanoparticles using carob leaf extract: Characterization and analysis of toxic effects in model organism Galleria mellonella L. (The greater wax moth).

Silver nanoparticles (Ag NPs) have been used in many studies due to their inhibitory properties on microorganisms such as bacteria and viruses. In recent years, due to global problems such as environmental pollution, the green synthesis (biosynthesis) method is frequently preferred because it is simple and low cost and does not require the use of toxic substances. The aim of this study is to synthesize silver nanoparticles (Ag NPs) from Ceratonia siliqua L. leaves and investigate their antioxidant and immunotoxic properties using Galleria mellonella last instar larvae. The UV spectrophotometer, TEM, XRD and FTIR measurements were used to characterize the Ag NPs. In this study, it was determined that the effects on antioxidant enzyme activities (SOD, CAT, GPx, GST), acetylcholinesterase (AChE) and total hemocyte count (THC) as well as phenoloxidase activity determine their effect on antioxidant defence and the immune system in model organism G. mellonella larvae. We observed that green synthesized Ag NPs accumulate in the midgut of the larvae and led to the increasing of CAT and SOD activities. GST and AChE activities were increased in the fat body of the larvae; otherwise, it was decreased in the midgut. Moreover, increases were found in THC and phenoloxidase activity. Consequently, green synthesized silver nanoparticles led to oxidative stress and immunotoxic effects on G. mellonella larvae.

Neuroprotective effect of Lannea egregia Alkaloid-rich leaf extracts in streptozotocin-induced diabetic rats

BackgroundStudies suggest that medicinal plant extracts can help reduce the neuron degeneration associated with diabetes. In this study, the neuroprotective effect of the alkaloid-rich extract from the leaves of Lannea egregia was assessed in rats with diabetes induced by streptozotocin (STZ). MethodsLannea egregia alkaloid-rich analysis was carried out via a known procedure. The rats were randomly assigned into five treatment groups (n = 8): normal control, diabetic-induced rats (45 mg/kg STZ), and diabetic rats treated with low doses of Lannea egregia leaf alkaloid-rich extract (50 mg/kg b.w, LEL) and high (100 mg/kg b.w, LEH) (300 mg/kg and 150 mg/kg), and metformin (200 mg/kg). On 22nd day of the experiment, animals were sacrificed, and their blood and brains were collected for neuro-biomarker analysis. ResultsDiabetic-induced rats that received metformin, LEL and LEH exhibited considerably reduced levels of dopamine, serotonin, norepinephrine, NO, MDA, and AChE, BChE activities when compared to untreated diabetic animals. Additionally, rats with diabetes that received treatment with metformin, LEL and LEH displayed a noticeable increase in ENTPDase, Na/K ATPase, GST, CAT, GPx, and SOD activities when compared to the untreated diabetic rats. Histological examination revealed improved brain architecture in the treated groups in contrast to those in diabetic-induced rats. ConclusionThe alkaloid-rich extracts of Lannea egregia might be effective in normalizing brain damage caused by complications of diabetes mellitus.

Tadalafil pretreatment attenuates doxorubicin-induced hepatorenal toxicity by modulating oxidative stress and inflammation in Wistar rats

Doxorubicin (DOX) is a widely used anticancer agent, but its clinical application is limited by significant off-target hepatorenal toxicity. Tadalafil (TAD), a selective phosphodiesterase-5 inhibitor used mainly for erectile dysfunction and pulmonary arterial hypertension, has shown potential in reducing oxidative stress. This study investigated TAD's chemoprotective effects and underlying mechanisms in DOX-induced hepatorenal toxicity in rats over 12 days. Eight groups of six rats each were orally pretreated with sterile water, silymarin (SIL), or TAD one hour before receiving intraperitoneal injections of 2.5 mg/kg DOX. On the 13th day, the rats were humanely sacrificed under inhaled halothane anesthesia, and serum was collected for hepatic and renal function tests, while liver and kidney tissues were analyzed for antioxidant enzyme activity, pro-inflammatory cytokines assay, and histopathological evaluation. DOX successfully induced hepatorenal toxicity, evidenced by significant increases (p<0.001, p<0.0001) in serum K+, urea, and creatinine levels, along with decreases in HCO3-, TCa2+, and Cl-. Tissue analysis showed reduced SOD, CAT, GST, and GPx activities, with elevated MDA and GSH levels. TAD pretreatment significantly ameliorated these biochemical alterations (p<0.05, p<0.001, p<0.0001), suggesting its potential as an effective chemoprophylactic adjuvant in the development of DOX-induced hepatorenal toxicity.

Biochemical responses of crayfish exposed to imidacloprid: assessment of oxidative stress, osmoregulatory response and neurotoxicity

ABSTRACT Among the nicotinoid insecticides, imidacloprid (IMI) may affect non-target invertebrates like aquatic crustaceans by entering the aquatic environment through rainfall. Nevertheless, there are not sufficient studies on the toxicity mechanism of IMI in crayfish. Here, we investigated the effects of IMI on oxidative stress, osmoregulatory, and neurotoxicity biomarkers. The 96-hour lethal concentration value (96 h-LC50) of IMI was determined to be 0.932 mg AI L−1. Four concentrations of IMI (0.932, 0.466, 0.233 and 0.117 mg AI L−1) were applied to crayfish for 96 h. Na+/K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, and total ATPase activities had almost significantly concentration-related inhibition in both gill and muscle tissues (p < 0.05). Compare with control in hepatopancreas, the activities of GST, SOD and GPx were increased, while the activities of AChE, CES and GR were decreased (p < 0.05). As non-enzymatic parameters, a statistically significant (p < 0.05) increase in MDA levels and a decrease in GSH levels were observed in the hepatopancreas compared to the control group, which was not related to the concentration. In conclusion, it can be said that the study's biomarkers are highly helpful and that commercially consumed crayfish are a significant non-target organism in the assessment of IMI toxicity.

Role of GSTP1 functional polymorphisms in molecular pathogenesis of colorectal cancer

Glutathione S-transferase P1 (GSTP1) plays a crucial role in the detoxification of harmful substances and cancer-causing agents. Single nucleotide polymorphisms (SNPs) in GSTP1 can affect its ability to catalyze reactions and detoxify, thereby influencing the risk of developing colorectal cancer (CRC). This study aimed to investigate the impact of functional SNPs (fSNPs) in GSTP1 on the risk of CRC, as well as their structural and functional consequences. We analyzed a total of 126 selected GSTP1 SNPs, including fSNPs rs1695 A > G (I105V) and rs1138272 C > T (A114V), in CRC patients (n = 103) and controls (n = 107) of south Indian origin using PCR-sequencing analysis with genomic DNA from blood samples. To assess the structural integrity of GSTP1 fSNPs, we conducted in silico analysis using various tools such as Swiss PDB Viewer, pyMOL mutagenesis wizard, ProSA-Web, and Pdbsum. Additionally, we performed functional characterization of GSTP1 fSNPs using cell and molecular biology techniques. Our findings revealed a significant association between the I105V fSNP and CRC risk, while the A114V fSNP did not show any significance. However, both fSNPs exhibited stronger linkage disequilibrium in patients compared to controls. In silico analysis indicated a loss of structural integrity and reduced electrostatic potential energy in the double mutant GSTP1 (V105/V114) compared to the native (I105/A114) or single mutant (V105/A114 and I105/V114) forms. Furthermore, FHC cells transfected with the GSTP1 I105V variant exhibited increased proliferation, invasion, and colony formation, along with decreased GST activity compared to carriers of the wild-type GSTP1. On the other hand, the GSTP1 A114V variant did not show a significant effect. Interestingly, FHC cells transfected with the double mutant GSTP1 variant (V105/V114) demonstrated synergistic and enhanced effects compared to the GSTP1 I105V variant. Consistent with these findings, plasma GST activity was significantly lower in haplogroups carrying both fSNPs compared to haplogroups with single or no fSNPs. To summarize, our findings indicate that while GSTP1 I105V alone contributes to the etiology of CRC, A114V does not; however, their combined presence has a more significant impact.

Heavy metals altered the xenobiotic metabolism of rats by targeting the GST enzyme: An in vitro and in silico study

Among all the heavy metals, Pb, Cd, and As are the most harmful pollutants in the environment. They reach into the organisms via various levels of food chains i.e. air and water. Glutathione-s-transferase (GST, E.C. 2.5.1.18), a key enzyme of xenobiotics metabolism, plays an important role in the removal of several toxicants. The present study aimed to evaluate any inhibitory action of these heavy metals on the GST enzyme isolated from the hepatic tissues of rats. A 10 % (w/v) homogenate of rat liver was prepared in cold and centrifuged at 4 °C at 9000xg for 30 min. The supernatant was collected and kept frozen at −20 °C or used fresh for carrying out different experiments. The activity of GST was monitored spectrophotometrically at 340 nm using 220 μg of soluble protein with varying equal substrate concentrations (0.125–2 mM) in phosphate buffer (50 mM, pH 6.5). To assess the impact of heavy metals on the enzyme activity, different concentrations of Cd (0–0.6 mM) and Pb (0–2 mM) were added to the reaction mixture followed by monitoring the residual activity. The optimum temperature and pH of rat liver GST were found to be 37 °C and 6.5, respectively. The Km value for GST was 0.69 mM and the Vmax was found to be 78.67 U/mg. The Cd and Pb significantly altered the kinetic behaviour of the enzyme. The Vmax and Kcat/Km parameters of GST were recorded to be decreased after interaction with Cd and Pb individually and showed a mixed type of inhibition pattern suggesting that these inhibitors may have a greater binding affinity either for the free enzyme or the substrate-enzyme complex. These metals showed a time-dependent enzyme inhibition profile. Cd was found to be the most potent inhibitor when compared to other treated metals; the order of inhibitory effect of metal ions was Cd>Pb>As. The in silico ion docking analysis for determining the probable interactions of Cd and Pb with fragmented GST validated that Cd exhibited higher inhibition potential for the enzyme as compared to Pb. The results of the present study indicated that exposure of both the Cd and Pb may cause significant inhibition of hepatic GST; the former with higher inhibitory potential than the later. However, As proved to be least effective against the enzyme under the aforesaid experimental conditions.

A multibiomarker approach to assess the effects of a BPA analogue-contaminated diet in the crab Carcinus aestuarii

Bisphenol A analogues are largely used plasticisers that are going to replace bisphenol A in many sectors. Due to this replacement, their discharge and presence in the marine coastal areas are increasing, with unknown consequences for organisms and the trophic chain. This study assessed the effects of three different bisphenols (BPAF, BPF and BPS) - alone or as a mixture - provided via food (exposed clams) to the crab Carcinus aestuarii. First, clams were exposed for two weeks to 300 ng/L of each of the three bisphenols and their mixture (100 ng/L of each) to allow the bioaccumulation of the contaminants in bivalves. Then, crabs were fed for two weeks with BPA analogue-exposed clams, while unexposed clams were used to feed control crabs. After 7 and 14 days, haemolymph, gills and hepatopancreas were collected from crabs to measure a battery of biomarkers indicative of cytotoxicity, oxidative stress and damage, neurotoxicity, physiological performance (respiration and excretion rate) and electron transport system activity. Lastly, bioaccumulation of BPA analogues was assessed by UHPLC-HRMS in crabs. Our findings revealed that BPA analogue-exposed clams were able to alter total haemocyte count, haemocyte size and their proliferation. The activity of immune enzymes, such as phosphatases and phenoloxidase was altered. Moreover, we observed an impairment of antioxidant and detoxifying enzymes like SOD, CAT, GST and GPX activities. Alterations of metabolism-involved enzymes and physiological parameters and increased oxidative damage to macromolecules like proteins, lipids, and DNA were also observed in crabs. Among BPA analogues, only bioaccumulation of BPAF, which has the highest Logkow value among the tested bisphenols, was evidenced in crabs. Overall, the obtained results indicated that crabs, under the tested experimental conditions at least, underwent alterations in cellular, biochemical and physiological responses following a diet of bisphenol-exposed clams, suggesting a potential ecotoxicological risk in the marine food chain.