Liver Toxicity Markers Research Articles

Silver Nanoparticles and L-Cysteine Composite Redresses Carbon Tetrachloride-Induced Hepatotoxicity in Swiss Albino Rats.

l-cysteine is a versatile amino acid that plays a pivotal role in synthesizing critical molecules, enzymatic catalysis, regulation, and electron transport. It also has tremendous potential to act as an adjuvant for enhancing the biological efficacy of various nanoparticles in vivo. The current study is aimed to evaluate the protective efficacy of silver nanoparticles (AgNPs) decorated with l-cysteine in carbon tetrachloride (CCl4)-induced hepatotoxicity in the Swiss albino rats as an animal model. The rats were divided into four treatment groups: Group 1 (control without any treatments), Group 2 treated with AgNPs and l-cysteine composite (5 mg/kg body weight on every third day), Group 3 (single dose of 1 mL/kg CCl4), and Group 4 treated with AgNPs-l-cysteine composite in the rats pre-administered with CCl4. After treatment for a month, the rats were killed, and their liver and blood samples were subjected to biochemical analysis and histological examination.: Group 2 showed all the parameters comparable to control Group 1. On the contrary, CCl4-treated, Group 3 rats showed abnormally raised liver function markers (AST and ALT) and liver toxicity markers (GGT, LDH, and total bilirubin) concomitant with disturbed oxidative stress parameters (GSH and MDA) compared to the control. However, Group 4 rats demonstrated a significant recovery from CCl4-induced biochemical alteration in the animals as compared to Group 3. In addition, the biochemical measurements were harmonious with the histological analysis of the liver sections of the treated rats. Hence, the proposed AgNPs-l-cysteine composite is a potent hepato-protecting agent in vivo that can be employed in regulating CCl4-induced hepatotoxicity or any drug or potential pharmaceutical compound exerting similar toxicity.

TROCEPT: An oncolytic virus platform engineered for αvß6 integrin targeted tumor-localised expression of an immune checkpoint inhibitor following intravenous delivery.

e14579 Background: TROCEPT is a novel tumor-selective replicating, oncolytic adenovirus type 5 (Ad5) rationally engineered to remove all natural tissue tropisms. This engineering addresses the main limitation of other viral therapies which infect normal tissues and are rapidly removed by the liver and are therefore limited to local delivery. TROCEPT has been further engineered to specifically bind to αvβ6 integrin which is expressed at high frequency in the majority of epithelial cancers, so that following intravenous delivery the virus targets tumor tissue where it replicates and amplifies. In addition, the TROCEPT platform encodes transgenes which enables in-tumor production of powerful therapeutic drugs. The lead programme, TROCEPT-01, encodes a fully human, full length immune checkpoint inhibitor (ICI) antibody. TROCEPT-01 is currently undergoing IND-enabling studies and is expected to enter First in Human studies in 2024 in multiple solid tumor indications. Methods: Proof of concept studies, both in vitro and in vivo, to demonstrate TROCEPT-01’s tumor selectivity and toxicity profile have been performed. Results: In vitro experiments demonstrate that viral replication, transgene expression and oncolytic cell death following infection with TROCEPT-01 is highly selective for αvβ6 integrin positive tumor cells compared to a panel of normal human primary cells. Intravenous delivery in in vivo SKOV3 ovarian human tumor xenograft mouse models demonstrate virus delivery, replication, and transgene expression in the tumor. In comparison to the parental Ad5 virus, the engineering of TROCEPT-01 leads to more viral delivery to the tumour, less delivery to normal tissue including liver, reduced markers of liver toxicity and reduced inflammatory cytokine release, all of which validate the tumour selectivity and safety profile of the virus. Conclusions: We have demonstrated that TROCEPT engineering enables highly selective tumor targeting and significant tumor exposure following intravenous delivery. TROCEPT-01’s in-tumor selective expression of ICIs enables high local drug concentration in the tumor, potentially reducing systemic toxicity and increasing efficacy. TROCEPT-01 is currently undergoing IND-enabling studies and is expected to enter First in Human studies in 2024 in multiple solid tumor indications.

MRNA Display Pipeline for Protein Biosensor Construction.

Despite the significant potential of protein biosensors, their construction remains a trial-and-error process. The most obvious approach for addressing this is to utilize modular biosensor architectures where specificity-conferring modalities can be readily generated to recognize new targets. Toward this goal, we established a workflow that uses mRNA display-based selection of hyper-stable monobody domains for the target of choice or ribosome display to select equally stable DARPins. These binders were integrated into a two-component allosteric biosensor architecture based on a calmodulin-reporter chimera. This workflow was tested by developing biosensors for liver toxicity markers such as cytosolic aspartate aminotransferase, mitochondrial aspartate aminotransferase, and alanine aminotransferase 1. We demonstrate that our pipeline consistently produced >103 unique binders for each target within a week. Our analysis revealed that the affinity of the binders for their targets was not a direct predictor of the binder's performance in a biosensor context. The interactions between the binding domains and the reporter module affect the biosensor activity and the dynamic range. We conclude that following binding domain selection, the multiplexed biosensor assembly and prototyping appear to be the most promising approach for identifying biosensors with the desired properties.

Impact of SMAASH-C, a novel nutritional supplement, on drug-seeking and toxicity in female and male rats.

Relapse to drug use after abstinence is a major challenge in treating substance use disorder. Exposure to drug-associated cues during abstinence can trigger intense craving and precipitate relapse. New and more effective anti-relapse interventions are critically needed, particularly for cocaine use disorder since no effective pharmacological intervention is available. We discovered that a nutritional supplement we developed as part of a nutritional approach for managing patients with substance use disorder reduced patient reports of drug craving and relapse. The goal of this study was to determine the efficacy of this supplement, SMAASH-C, at reducing drug-craving/relapse vulnerability in males and females in rat models with cocaine. Effects were determined following extended-access cocaine self-administration (24-hr/day for 10 days) and a two-week treatment regimen at a moderate and moderate-to-high dose (0.4 and 0.8 g/kg/day) as well as a 6-week regimen at a moderate dose (0.4 g/kg/day; Experiment 2). We also determined its efficacy to offset serum markers of organ toxicity in response to chronic cocaine self-administration and abstinence (aspartate transaminase, alanine transaminase, amylase; urea nitrogen). In females, both the 2- and 6-week SMAASH-C treatment regimens reduced cocaine-seeking (extinction or cue-induced reinstatement), particularly when drug-seeking was heightened (e.g., during estrus). Despite a lack of efficacy to reduce drug-seeking in males, SMAASH-C treatment normalized cocaine/abstinence-induced increases in serum levels of aspartate transaminase and amylase, which are markers of liver and pancreatic toxicity respectively. Thus, the beneficial effects of oral SMAASH-C treatment over abstinence following chronic cocaine self-administration appears to be sex-specific.

Targeting the PI3K/pAKT/mTOR/NF-κB/FOXO3a signaling pathway for suppressing the development of hepatocellular carcinoma in rats: Role of the natural remedic Suaeda vermiculata forssk.

Liver malignancy is well recognized as a prominent health concern, with numerous treatment options available. Natural products are considered a renewable source, providing inspiring chemical moieties that could be used for cancer treatment. Suaeda vermiculata Forssk has traditionally been employed for management of hepatic conditions, including liver inflammation, and liver cirrhosis, as well as to improve general liver function. The findings of our earlier study demonstrated encouraging in vivo hepatoprotective benefits against liver injury generated by paracetamol and carbon tetrachloride. Additionally, Suaeda vermiculata Forssk exhibited cytotoxic activities in vitro against Hep-G2 cell lines and cell lines resistant to doxorubicin. The present investigation aimed to examine the potential in vivo hepatoprotective efficacy of Suaeda vermiculata Forssk extract (SVE) against hepatocellular carcinoma induced by diethylnitrosamine (DENA) in rats. The potential involvement of the PI3K/AKT/mTOR/NF-κB pathway was addressed. Sixty adult male albino rats were allocated into five groups randomly (n = 10). First group received a buffer, whereas second group received SVE only, third group received DENA only, and fourth and fifth groups received high and low doses of SVE, respectively, in the presence of DENA. Liver toxicity and tumor markers (HGFR, p-AKT, PI3K, mTOR, NF-κB, FOXO3a), apoptosis markers, and histopathological changes were analyzed. The current results demonstrated that SVE inhibited PI3K/AKT/mTOR/NF-κB pathway as well as increased expression of apoptotic parameters and FOXO3a levels, which were deteriorated by DENA treatment. Furthermore, SVE improved liver toxicity markers and histopathological changes induced by DENA administration. This study provided evidence for the conventional hepatoprotective properties attributed to SV and investigated the underlying mechanism by which its extract, SVE, could potentially serve as a novel option for hepatocellular carcinoma (HCC) treatment derived from a natural source.

Silver Nanoparticles Decorated with Curcumin Enhance the Efficacy of Metformin in Diabetic Rats via Suppression of Hepatotoxicity.

Hepatotoxicity is one of the significant side effects of chronic diabetes mellitus (DM) besides nephrotoxicity and pancreatitis. The management of this disease is much dependent on the restoration of the liver to its maximum functionality, as it is the central metabolic organ that gets severely affected during chronic diabetes. The present study investigates if the silver nanoparticles decorated with curcumin (AgNP-Cur) can enhance the efficacy of metformin (a conventional antidiabetic drug) by countering the drug-induced hepatoxicity. Swiss albino rats were categorized into six treatment groups (n = 6): control (group I without any treatment), the remaining five groups (group II, IV, V, VI) were DM-induced by streptozocin. Group II was untreated diabetic positive control, whereas groups III was administered with AgNP-cur (5 mg/kg). Diabetic group IV treated with metformin while V and VI were treated with metformin in a combination of the two doses of NPs (5 and 10 mg/kg) according to the treatment schedule. Biochemical and histological analysis of blood and liver samples were conducted after the treatment. The groups V and VI treated with the combination exhibited remarkable improvement in fasting glucose, lipid profile (HDL and cholesterol), liver function tests (AST, ALT), toxicity markers (GGT, GST and LDH), and redox markers (GSH, MDA and CAT) in comparison to group II in most of the parameters. Histological evaluation and comet assay further consolidate these biochemical results, pleading the restoration of the cellular structure of the target tissues and their nuclear DNA. Therefore, the present study shows that the NPs can enhance the anti-diabetic action by suppression of the drug-mediated hepatoxicity via relieving from oxidative stress, toxic burden and inflammation.

Design and synthesis of new 1,2,3-triazoles derived from eugenol and analogues with in vitro and in vivo activity against Trypanosoma cruzi

Chagas disease (CD) is a neglected tropical disease endemic in 21 countries and affects about 8 million people around the world. The pharmacotherapy for this disease is limited to two drugs (Benznidazole and Nifurtimox) and both are associated with important limitations, as low cure rate in the chronic phase of the disease, high toxicity and increasing resistance by Trypanosoma cruzi. Recently, we reported a bioactive 1,2,3-triazole (compound 35) active in vitro (IC50 42.8 μM) and in vivo (100 mg/kg) against T. cruzi Y strains and preliminary in silico studies suggested the cysteine protease cruzain as a possible target. Considering these initial findings, we describe here the design and synthesis of new 1,2,3-triazoles derivatives of our hit compound (35). The triazoles were initially evaluated against healthy cells derived from neonatal rat cardiomyoblasts (H9c2 cells) to determine their cytotoxicity and against epimastigotes forms of T. cruzi Y strain. The most active triazoles were compounds 26 (IC50 19.7 μM) and 27 (IC50 7.3 μM), while benznidazole was active at 21.6 μM. Derivative 27 showed an interesting selectivity index considering healthy H9c2 cells (>77). Promising activities against trypomastigotes forms of the parasite were also observed for triazoles 26 (IC50 20.74 μM) and 27 (IC50 8.41 μM), mainly 27 which showed activity once again higher than that observed for benznidazole (IC50 12.72 μM). While docking results suggested cruzain as a potential target for these compounds, no significant enzyme inhibition was observed in vitro, indicating that their trypanocidal activity is related to another mode of action. Considering the promising in vitro results of triazoles 26 and 27, the in vivo toxicity was initially verified based on the evaluation of behavioral and physiological parameters, mortality, effect in body weight gain, and through the measurement of AST/ALT enzymes, which are markers of liver toxicity. All these evaluations pointed to a good tolerability of the animals, especially considering triazole 27. A reduction in parasitemia was observed among animals treated with triazole 27, but not among those treated with derivative 26. Regarding the dosage, derivative 27 (100 mg/kg) was the most active sample against T. cruzi infection, showing a 99.4% reduction in parasitemia peak. Triazole 27 at a dosage of 100 mg/kg influenced the humoral immune response and reduced myocarditis in the animals, bringing antibody levels closer to those observed among healthy mice. Altogether, our results indicate compound 27 as a new lead for the development of drug candidates to treat Chagas disease.

Effects of Exercise Preconditioning on Doxorubicin-Induced Liver and Kidney Toxicity in Male and Female Rats.

Doxorubicin (DOX) is a highly effective chemotherapy agent prescribed for cancer treatment. However, the clinical use of DOX is limited due to off-target toxicity in healthy tissues. In this regard, hepatic and renal metabolic clearance results in DOX accumulation within these organ systems. Within the liver and kidneys, DOX causes inflammation and oxidative stress, which promotes cytotoxic cellular signaling. While there is currently no standard of care to treat DOX hepatic- and nephrotoxicity, endurance exercise preconditioning may be an effective intervention to prevent elevations in liver alanine transaminase (ALT) and aspartate aminotransferase (AST) and to improve kidney creatinine clearance. To determine whether exercise preconditioning is sufficient to reduce liver and kidney toxicity resulting from acute exposure to DOX chemotherapy treatment, male and female Sprague-Dawley rats remained sedentary or were exercise trained prior to saline or DOX exposure. Our findings demonstrate that DOX treatment elevated AST and AST/ALT in male rats, with no effects of exercise preconditioning to prevent these increases. We also showed increased plasma markers of renin-angiotensin-aldosterone system (RAAS) activation and urine markers of proteinuria and proximal tubule damage, with male rats revealing greater differences compared to females. Exercise preconditioning showed improved urine creatinine clearance and reduced cystatin c in males, while females had reduced plasma angiotensin II (AngII) levels. Our results demonstrate both tissue- and sex-specific responses related to the effects of exercise preconditioning and DOX treatment on markers of liver and kidney toxicity.

Hepatoprotective effect of the methanol extract of Luffa cylindrica fruit on carbon-tetrachloride induced chronic liver injury

Luffa cylindrica (Linn) commonly called sponge gourds has both medicinal and nutritional properties. It is used traditionally for the management of liver diseases. Herein, we investigated the hepatoprotective effect of the crude methanol extract of L. cylindrica fruit in rats chronically exposed to carbon tetrachloride (CCl4). Male rats were exposed to CCl4 twice a week for six weeks and the extract was administered five times a week for six weeks. Markers of liver toxicity, antioxidant enzymes and liver peroxidation were evaluated and histological analysis of the liver was carried out. Significant reduction in serum markers (ALT, AST and ALP), increase in antioxidant enzyme and reduction in lipid peroxidation compared to CCl4 were observed in rats exposed to both CCl4 and the extract. CCl4-induced liver lesions were ameliorated by the extract. These show the protective effect of the methanol extract of L. cylindrica on CCl4-induced chronic liver injury in rats.

Chemical Composition of Various Nepeta cataria Plant Organs' Methanol Extracts Associated with In Vivo Hepatoprotective and Antigenotoxic Features as well as Molecular Modeling Investigations.

This report summarizes the chemical composition analysis of Nepeta cataria L. flower, leaf, and stem methanol extracts (FME, LME, SME, respectively) as well as their hepatoprotective and antigenotoxic features in vivo and in silico. Herein, Wistar rat liver intoxication with CCl4 resulted in the generation of trichloromethyl and trichloromethylperoxy radicals, causing lipid peroxidation within the hepatocyte membranes (viz. hepatotoxicity), as well as the subsequent formation of aberrant rDNA adducts and consequent double-strand break (namely genotoxicity). Examined FME, LME, and SME administered orally to Wistar rats before the injection of CCl4 exerted the most notable pharmacological properties in the concentrations of 200, 100, and 50 mg/kg of body weight, respectively. Thus, the extracts’ hepatoprotective features were determined by monitoring the catalytic activities of enzymes and the concentrations of reactive oxidative species, modulating the liver redox status. Furthermore, the necrosis of hepatocytes was assessed by means of catalytic activities of liver toxicity markers. The extracts’ antigenotoxic features were quantified using the comet assay. Distinct pharmacological property features may be attributed to quercitrin (8406.31 μg/g), chlorogenic acid (1647.32 μg/g), and quinic acid (536.11 μg/g), found within the FME, rosmarinic acid (1056.14 μg/g), and chlorogenic acid (648.52 μg/g), occurring within the LME, and chlorogenic acid (1408.43 μg/g), the most abundant in SME. Hence, the plant’s secondary metabolites were individually administered similar to extracts, upon which their pharmacology in vivo was elucidated in silico by means of the structure-based studies within rat catalase, as a redox marker, and rat topoisomerase IIα, an enzyme catalyzing the rat DNA double-strand break. Conclusively, the examined N. cataria extracts in specified concentrations could be used in clinical therapy for the prevention of toxin-induced liver diseases.

Comparison of DLin-MC3-DMA and ALC-0315 for siRNA Delivery to Hepatocytes and Hepatic Stellate Cells.

Ionizable cationic lipids are essential for efficient in vivo delivery of RNA by lipid nanoparticles (LNPs). DLin-MC3-DMA (MC3), ALC-0315, and SM-102 are the only ionizable cationic lipids currently clinically approved for RNA therapies. ALC-0315 and SM-102 are structurally similar lipids used in SARS-CoV-2 mRNA vaccines, while MC3 is used in siRNA therapy to knock down transthyretin in hepatocytes. Hepatocytes and hepatic stellate cells (HSCs) are particularly attractive targets for RNA therapy because they synthesize many plasma proteins, including those that influence blood coagulation. While LNPs preferentially accumulate in the liver, evaluating the ability of different ionizable cationic lipids to deliver RNA cargo into distinct cell populations is important for designing RNA-LNP therapies with minimal hepatotoxicity. Here, we directly compared LNPs containing either ALC-0315 or MC3 to knock-down coagulation factor VII (FVII) in hepatocytes and ADAMTS13 in HSCs. At a dose of 1 mg/kg siRNA in mice, LNPs with ALC-0315 achieved a 2- and 10-fold greater knockdown of FVII and ADAMTS13, respectively, compared to LNPs with MC3. At a high dose (5 mg/kg), ALC-0315 LNPs increased markers of liver toxicity (ALT and bile acids), while the same dose of MC3 LNPs did not. These results demonstrate that ALC-0315 LNPs achieves potent siRNA-mediated knockdown of target proteins in hepatocytes and HSCs, in mice, though markers of liver toxicity can be observed after a high dose. This study provides an initial comparison that may inform the development of ionizable cationic LNP therapeutics with maximal efficacy and limited toxicity.

Effectiveness of "Moro" Blood Orange Citrus sinensis Osbeck (Rutaceae) Standardized Extract on Weight Loss in Overweight but Otherwise Healthy Men and Women-A Randomized Double-Blind Placebo-Controlled Study.

This study aimed to assess the efficacy of a blood orange Citrus sinensis standardized extract from “Moro” cultivar, on weight loss in overweight but otherwise healthy individuals. Anthocyanins and particularly cyanidin 3-glucoside, found in a large variety of fruits including Sicilian blood oranges, can help to counteract weight gain and to reduce body fat accumulation through the modulation of antioxidant, anti-inflammatory and metabolic pathways. In this randomized, double blind, placebo-controlled study, all participants (overweight adults aged 20–65 years old) were randomized to receive either Moro blood orange standardized extract or a placebo daily for 6-months. The primary outcome measure was change in body mass and body composition at the end of the study. After 6-months, body mass (4.2% vs. 2.2%, p = 0.015), body mass index (p = 0.019), hip (3.4 cm vs. 2.0 cm, p = 0.049) and waist (3.9 cm vs. 1.7 cm, p = 0.017) circumferences, fat mass (p = 0.012) and fat distribution (visceral and subcutaneous fat p = 0.018 and 0.006, respectively) were all significantly better in the extract supplemented group compared to the placebo (p < 0.05). In addition, all safety markers of liver toxicity were within the normal range throughout the study for both analyzed groups. Concluding, the present study demonstrates that Moro blood orange standardized extract may be a safe and effective option for helping with weight loss when used in conjunction with diet and exercise.

LDL-Based Lipid Nanoparticle Derived for Blood Plasma Accumulates Preferentially in Atherosclerotic Plaque.

Apolipoprotein-based drug delivery is a promising approach to develop safe nanoparticles capable of targeted drug delivery for various diseases. In this work, we have synthesized a lipid-based nanoparticle (NPs) that we have called “Aposomes” presenting native apolipoprotein B-100 (apoB-100), the primary protein present in Low-Density Lipoproteins (LDL) on its surface. The aposomes were synthesized from LDL isolated from blood plasma using a microfluidic approach. The synthesized aposomes had a diameter of 91 ± 4 nm and a neutral surface charge of 0.7 mV ± mV. Protein analysis using western blot and flow cytometry confirmed the presence of apoB-100 on the nanoparticle’s surface. Furthermore, Aposomes retained liposomes’ drug loading capabilities, demonstrating a prolonged release curve with ∼80% cargo release at 4 hours. Considering the natural tropism of LDL towards the atherosclerotic plaques, we evaluated the biological properties of aposomes in a mouse model of advanced atherosclerosis. We observed a ∼20-fold increase in targeting of plaques when comparing aposomes to control liposomes. Additionally, aposomes presented a favorable biocompatibility profile that showed no deviation from typical values in liver toxicity markers (i.e., LDH, ALT, AST, Cholesterol). The results of this study demonstrate the possibilities of using apolipoprotein-based approaches to create nanoparticles with active targeting capabilities and could be the basis for future cardiovascular therapies.

Phytochemical Characterization, Hepatoprotective Activity on Alcohol-Induced Toxicity of the Aqueous Extract of Curcuma longa (Zingiberaceae) in Wistar Rats

Introduction: Liver or hepatic disease refers to different conditions that affect the liver. Chronic alcohol consumption is one of the most frequent causes of liver disease and accounts for about 55% of liver cirrhosis deaths recorded in Cameroon in 2020. Standard accessible treatments focus on end-stage liver disease with safety and efficacy obstacles. We have a research gap in Cameroon to understand the alternative use of natural products as treatment with a long traditional history of safe use. Curcuma longa has long been a source of traditional and modern medicine. It is commonly used in Cameroon as a spice and herbal product with some level of activity against various forms of liver disease.&#x0D; Objective: To phytochemically screen for bioactive metabolites and evaluate the hepatoprotective activity of the aqueous extract of Curcuma longaon alcohol-induced toxicity in Wistar rats.&#x0D; Methods: Phytochemical screening was carried out on the aqueous extract obtainedfrom maceration of plant rhizomes. Three doses (125, 250 and 500mg/Kg) of the plant extract and the reference (Silymarin 50mg/Kg) were administered daily (p.o) to rats 30 min before administration of 40% alcohol (2mL/100g p.o) for 21 days. Biochemical parameters such as ALAT, ASAT, GGT, Bilirubin and Lipid profile were quantified and histological studies of the liverwas carried out using standard procedures.&#x0D; Results: Phytochemical screening of the aqueous extract of C. longa revealed polyphenols such as flavonoids, tannins, quinones, saponins and phlobatanins. The plant showed hepatoprotective activity by decreasing liver toxicity markers such as ASAT, ALAT, GGT and Bilirubin. Histology revealed dose-dependent protection with 500 mg/Kg showing the most cellular integrity, no central vein occlusion and minimal fibrosis.&#x0D; Conclusion: This study indicated the presence of polyphenols like flavonoids and tannins in the aqueous extract of C. longa. The presence of these secondary metabolites in the studied extract justifies its antioxidant and anti-inflammatory properties confirmed by its hepatoprotective effects on alcohol-induced toxicity. This was clearly shown by biochemical and histological parameters. More sensitive and specific methods are required to test for these secondary metabolites in serum.

Toxicological Profile of Umbilical Cord Blood-Derived Small Extracellular Vesicles.

The development and adoption of cell therapies has been largely limited by difficulties associated with their safety, handling, and storage. Extracellular vesicles (EV) have recently emerged as a likely mediator for the therapeutic effect of cells, offering several advantages over cell therapies. Due to their small size and inability to expand and metastasize, EV are generally considered safer than cell transplantation. Nevertheless, few studies have scrutinized the toxicity profile of EV, particularly after repeated high-dose administration. The present study aimed to evaluate a preparation of small EV obtained from umbilical cord blood mononuclear cells (UCB-MNC-sEV) for its cytotoxicity in different cell lines, as well as its differential accumulation, distribution, and toxicity following repeated intravenous (IV) administrations in a rodent model. In vitro, repeated sEV exposure in concentrations up to 1 × 1011 particles/mL had no deleterious impact on the viability or metabolic activity of peripheral blood mononuclear cells, THP-1 monocytes, THP-1-derived macrophages, normal dermal human fibroblasts, or human umbilical vein endothelial cells. DiR-labelled sEV, injected intravenously for four weeks in healthy rats, were detected in clearance organs, particularly the kidneys, spleen, and liver, similarly to control dye. Moreover, repeated administrations for six and twelve weeks of up to 1 × 1010 total particles of sEV dye were well-tolerated, with no changes in general haematological cell counts, or kidney and liver toxicity markers. More importantly, unlabelled sEV likewise did not induce significant alterations in cellular and biochemical blood parameters, nor any morphological changes in the heart, kidney, lung, spleen, or liver tissue. In sum, our data show that UCB-MNC-sEV have no significant toxicity in vitro or in vivo, even when administered repeatedly at high concentrations, therefore confirming their safety profile and potential suitability for future clinical use.

Evaluation of the Systemic Serum Exposure and Acute Toxicity of the Aqueous Extract of Curcuma longa (Zingiberaceae) Rhizomes in Wistar Rats

Introduction: Liver toxicity has become a public health concern as more people globally get exposed to xenobiotics with the potential to cause liver damage and consequent liver cirrhosis. The increase in liver toxicant abuse has necessitated the exploration of xenobiotic exposure levels when addressing therapeutic measures using alternative herbal remedies. The increasing use of herbal products as alternative therapy needs regulatory alignment through evidence-based support for the safety and efficacy of these natural products. To undertake preclinical discovery of new metabolites from medicinal products, the objective of this study was to investigate the systemic serum exposure and acute toxicity of the aqueous extract of Curcuma longa (Zingiberaceae) rhizomes on Wistar rat models.&#x0D; Methods: Phytochemical screening was carried out on the aqueous extract obtained by maceration of the dried plant rhizomes. Standard screening techniques for plant metabolites were used to screen blood serum after animal exposure with the extract. After a 500mg/Kg dose, systemic exposure was evaluated in blood samples collected at 30-minute intervals for one hour.&#x0D; For acute toxicity, a single 2000mg/Kg by body weight dose of the plant extract and the reference (Silymarin 50mg/Kg) were administered to rats, and they were observed for 14 days. Biochemical markers of toxicity such as ALAT, ASAT, GGT, Bilirubin were quantified, and histological studies of the liver were carried out.&#x0D; Results: No secondary metabolites were identified at 30 mins and 1hr in rat serum following a 500 mg/Kg oral dose. Administration of a 2000 mg/Kg oral dose to rats was well tolerated, and there were no deaths or significant target organ toxicity. The plant showed no lethality at the dose of 2000mg/kg body weight and decreased liver toxicity markers such as ASAT, ALAT, GGT, and Bilirubin. Histology revealed no significant damage to liver hepatocytes, no central vein occlusion, and no evidence of fibrosis.&#x0D; Conclusion: There were no systemically available secondary metabolites at a dose of 500 mg/Kg after the qualitative screening; more sensitive and specific methods are required to test these secondary metabolites in serum. This study confirmed the safety margin of Curcuma longa with no lethality following a single oral dose of 2000mg/Kg and after observation for 14 days. There was a low expression of biochemical markers of toxicity ALAT, ASAT, and no histological indication of liver damage.

Antioxidant-rich fraction of Amomum subulatum fruits mitigates experimental methotrexate-induced oxidative stress by regulating TNF-α, IL-1β, and IL-6 proinflammatory cytokines.

The culinary spice Amomum subulatum was assessed for its phytochemical composition, in vitro antioxidant potential, and in vivo ameliorating effect against methotrexate (MTX)-induced toxicities. Phytochemical analysis of methanolic extract of A.subulatum dry fruits (MEAS) confirmed the presence of different bioactive secondary metabolites. MEAS scavenged reactive free radicals and inhibited lipid peroxidation in vitro. To confirm the antioxidant efficiency of MEAS, in vivo experiment was carried out in which MTX was administered to induce oxidative stress. Co-administration of MEAS reduced MTX-induced hepatic, renal, and pulmonary toxicities via significantly (p<.01) enhancing antioxidant status and reducing oxidative stress. MTX treatment significantly (p<.01) increased liver and kidney toxicity markers and increased proinflammatory cytokine (TNF-α, IL-1β, and IL-6) levels. However, co-administration of MEAS significantly (p<.01) reduced their levels, and tissue histopathology confirmed the protective effect of MEAS in maintaining normal tissue architecture following MTX treatment. Protective effect of MEAS is accredited to the antioxidant and anti-inflammatory properties exhibited by bioactive compounds in MEAS. PRACTICAL APPLICATIONS: Amomum subulatum (Black cardamom) is a folkloric and culinary spice used for its organoleptic, nutritional, and medicinal properties. This study demonstrated the phytochemical composition and antioxidant potential of methanolic extract of A.subulatum dry fruits (MEAS). Toxicities associated with MTX therapy limit its clinical application. MEAS attenuated methotrexate-induced oxidative stress, inflammation, and associated organ damages, suggesting the possible therapeutic application of A.subulatum in reducing oxidative stress and associated diseases. Our results showed that A.subulatum is a potential functional food, which may be used for the betterment of health due to its richness in antioxidant and anti-inflammatory phytochemicals.

Modulation of immune mechanisms during hepatoprotective effects of Dawa-Ul-Kurkum, a Unani polyherbal preparation in experimental model of paracetamol induced liver damage in rats

Objective: Iimmunomodulatory effects of Dawa-Ul-Kurkum, a Unani polyherbal preparation and the possible mechanisms in experimental model of paracetamol induced liver damage in rats.&#x0D; Materials and methods: The drug Dawa-Ul- Kurkum has been prepared and provided by CRIUM, Hyderabad. Silymarin was purchased from Sigma-Aldrich (USA) and paracetamol (marketed by Cipla LTD) purchased from general pharmacy shop. Biochemical kits were purchased from ERBA Diagonostic Mannheim Gmbh. Serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST) and serum alkaline phosphatase (ALP) were estimated by Kinetic method of International Federation of Clinical Chemistry (IFCC), serum bilirubin and total protein were estimated by End Point assay as per the instruction of the Kit Manufacture’s manual.Immunoglobulin and cytokine levels were assessed by ELISA kit manual method. Delayed type hypersensitivity reaction by Institoris et al method. Lipid peroxidation is measured spectrophotometrically as 2-thiobarbituric acid-reactive substance (TBARS), Glutathione (GSH) levels were estimated by the method of Ellman and NOx concentrations were determined by using the Griess reaction as described previously by Tracey et al.&#x0D; Results: Liver damage was induced in Wistar rats by administration of paracetamol and the effects of various drug treatments were assessed on morphological, biochemical, immunoglobulin, cytokine level and histological markers of liver toxicity. In the vehicle treated experimental group, administration of paracetamol induced significant derangements in liver function as evidenced by increased levels of SGOT, SGPT, alkaline phosphatase bilirubin and decrease in total protein, and reductions in body weight and increased liver weights as compared to controls. Histopathological examination showed Periportal necrosis with haemorrhages in experimental control. Pretreatment with Dawa-Ul-Kurkum (DK, 250 and 500 mg/kg) and hydroalcoholic extract of DK (HA, 500 and 1000 mg/kg) showed protective effects against the paracetamol induced biochemical, immunoglobulin, cytokine, delayed type hypersensitivity reaction and histopathological derangements of liver function following paracetamol. Conclusion: Both DK and its 50% hydro-alcoholic extract were found to be effective against paracetamol induced liver damage as they significantly prevented the hepatotoxic damage induced in rats.

Toxicological implications of sequential administration of herbal and conventional medicines: Evidence from an in vivo study on Azadirachta indica and artesunate in male Wistar rats

In most parts of West Africa and other developing countries, herbal medicines are sometimes used by patients concomitantly receiving conventional drugs, which can result in potentially serious adverse effects. This study examined in vivo cytotoxic effects of Azadirachta indica extracts followed by artesunate administration on some markers of liver and kidney toxicity. Serum ALT, GGT, urea, creatinine, interleukin 1β, tumor necrosis factor α, tissue malondialdehyde and glutathione levels and liver and kidney histology in healthy male Wistar rats administered 100 and 200 mg/kg A. indica for 5 days followed by 10 mg/kg Artesunate for 5 days was determined. Results showed significantly ( p &lt; 0.05) higher serum ALT, GGT, urea, creatinine, interleukin 1β and tumor necrosis factor α levels with proportional increase of 16.5, 21.7, 9.2, 6.9, 9.1 and 9.1% respectively when compared to normal control was observed. Malondialdehyde levels were significantly ( p &lt; 0.05) higher with a proportional increase of 57.8%, while glutathione levels were significantly ( p &lt; 0.05) lower with a proportional decrease of 13.4% in liver homogenates of the treated rats relative to normal control. Histological examination of the liver and kidney of the co-treated rats showed vascular congestion and necrosis. Collectively, the results suggest that administration of A. indica followed by artesunate could predispose to liver and kidney associated cytotoxicity. These findings could have implications for people who habitually use herbal preparations and conventional drugs in sequential fashion.

Methanolic Extract of Muntingia Calabura L. Mitigates 1,2-Dimethyl Hydrazine Induced Colon Carcinogenesis in Wistar Rats

Objective The present study aimed to evaluate the efficacy of methanolic extract of Muntingia calabura L. leaves (MEMC) in ameliorating oxidative stress and inflammation associated with 1,2-dimethyl hydrazine (DMH) induced colon cancer. Methods The antioxidant enzymes, oxidative stress markers, liver and renal toxicity markers were evaluated. Histopathological examination of colon tissues was carried out with the aid of alcian blue stain and Hematoxylin and Eosin stain. Results MEMC supplementation at doses of 100 and 200 mg/kg body weight of rats causes the antioxidant enzymic levels to retain near to its normal range. Meanwhile the oxidative stress markers, which showed an elevation from its normal level upon DMH administration, gets significantly reduced on MEMC treatment. Histopathological observation also revealed that the severity of colorectal cancer was reduced by the supplementation of MEMC. Conclusion The findings from the present study showed that MEMC can exert a potential role to ameliorate the oxidative stress and inflammation associated with colorectal cancer.