Levels Of Marker Enzymes Research Articles

To investigate the expression of miRNA in nicotine induced GISTs tumor rats and the mitigation of cardiovascular complications

The common for the most part of tumor (GISTs) Gastrointestinal stromal tumor is known mesenchymal neoplasms of the tumor in gastrointestinal tract. Detection of serum miRNA 1, 15b, 21 adjustment mainly cardio marker enzymes and their mechanism of action and heart tissue, observed changes in LDH, CK-MB, AST, ALT, lipid peroxidation (LPO) and Troponin T estimations/biochemical changes observed in serum of experimental subjects (nicotine treated rats, 0.6 mg/kg, i.p.). In the present study, gastrointestinal stromal tumors of rats and the mitigation of cardiovascular complications was experimentally induced in rats by nicotine i.p. treatment of adult rats and maintained control rats. Confirm significantly decreased expression of miRNA-1 (nicotine induced gastrointestinal tumor rats), the levels of miRNA-15b expression is decrease in group II but miRNA-21 expression values are increased in experimental rats (group II) with control rats (group I). Body weight decreased significantly in group II nicotine induced rats and heart weight increased in group II but systolic blood pressure is decreased in experimental rats (group II) with control rats. Cardio marker enzyme levels were measured and increases LDH levels, slight raises CK-MB concentration range in group II. AST, ALT marker enzyme levels are higher in experimental rats with controls, troponin T values are clearly declare that some cardio complications in group II. Observed Ldh, Ck-Mb, Ast and Alt levels are decreased followed by antioxidant enzymes SOD, catalase but lipidperoxidation and protein carbonyls values are increased in nicotine induced rats and match up to normal rats. GST levels are increases significantly and Gpx, GRd and GSH levels are decreased in group II nicotine induced rats and corporately control group I. This study to investigates, for the first time, the effect of nicotine i.p., in adult rats (nicotine treated) and control rats. Reports of serum as well as heart tissue adjustment in molecular levels.

Catechism of protective potential of Areca catechu in Isoprenaline challenged Wistar rats

Areca catechu is an important ancient drug commonly known as Supari in ayurvedic system of medicine. A lot of research work has been done on Areca catechu regarding various cardiovascular disorders such as Hypertension, Arrhythmia but no work has been done to find out its cardioprotective activity. Experimental procedures done on Wistar Albino rats as Normal control group (NC) received 0.5ml of normal saline throughout the study and served as control. Isoprenaline group (ISO) received 0.5ml of normal saline throughout the experimental phase and received Isoprenaline (85mg/kg, s.c.) on 14th and 15th day at a time lapse of 24 hours. Standard group (STD) received Metoprolol (pure) (10mg/kg/day, p.o.) for 13 days and received Isoprenaline (85mg/kg, s.c.) on 14th and 15th day. Test group received Areca catechu extract (100mg/kg/day, p.o.) and (200 mg/kg/day, p.o.) respectively for 13 days and Isoprenaline (85mg/kg, s.c.) on 14th and 15th day. On 16thday animals were sacrificed. The level of marker enzyme in serum as AST, ALT, CK, LDH, Troponin-I have shown significant decrease (P<0.001) in rats pre-treated with Areca catechu when compared to toxic group. Further, histopathological examination showed the reduction of necrosis, edema and inflammation following Areca catechu pre-treatment. Findings revealed that Areca catechu may be a potential preventive and therapeutic agent against the myocardial necrosis associated ischemic heart disease. Thus, the aqueous ethanolic Areca catechu seeds extract could be recommended as a potential cardioprotective drug.

Nerolidol, a Sesquiterpene from the Essential Oils of Aromatic Plants, Attenuates Doxorubicin-Induced Chronic Cardiotoxicity in Rats.

The clinical usage of doxorubicin (DOX), a potent anthracycline antineoplastic drug, is limited due to its cardiotoxicity. The aim of this study was to assess the possible cardioprotective effects of nerolidol (NERO) in a rat model of DOX-induced chronic cardiotoxicity and the underlying molecular mechanisms. DOX (2.5 mg/kg) was injected intraperitoneally once in a week for 5 weeks to induce chronic cardiotoxicity in male albino Wistar rats. The rats were treated with NERO (50 mg/kg, orally) 6 days a week for a duration of 5 weeks. DOX-injected rats showed a significant decline in cardiac function, elevated levels of serum cardiac marker enzymes, and enhanced oxidative stress markers along with altered PI3K/Akt and Nrf2/Keap1/HO-1 signaling pathways. DOX also triggered the activation of NF-κB/MAPK signaling and increased the levels/expression of proinflammatory cytokines (TNF-α, IL-6, and IL-1β) and expression of inflammatory mediators (iNOS and COX-2) in the heart. DOX activated NLRP3 inflammasome-mediated pyroptotic cell death along with fibrosis, mitochondrial dysfunction, DNA damage, and apoptosis in the myocardium. Additionally, histological studies, TUNEL staining, and myocardial lesions revealed structural alterations of the myocardium. NERO treatment showed considerable protective effects on the biochemical and molecular parameters studied. The findings demonstrate that NERO protects against DOX-induced chronic cardiotoxicity and the observed cardioprotective effects are attributed to its potent antioxidant and free radical scavenging properties.

Nerolidol Attenuates Oxidative Stress, Inflammation, and Apoptosis by Modulating Nrf2/MAPK Signaling Pathways in Doxorubicin-Induced Acute Cardiotoxicity in Rats.

The clinical usage of doxorubicin (DOX), a potent anthracycline antineoplastic drug, is often limited by its cardiotoxic effects. Thus, for improving usage of DOX, the aim of this study was to assess the cardioprotective effects of nerolidol (NERO) in a rat model of DOX-induced acute cardiotoxicity and examine underlying molecular mechanisms that contribute to these effects. To induce acute cardiotoxicity male albino Wistar rats were injected with single dose intraperitoneal DOX (12.5 mg/kg). The rats were treated with NERO (50 mg/kg, orally) for five days. DOX-injected rats showed elevated levels of cardiac marker enzymes and enhanced oxidative stress markers along with altered Nrf2/Keap1/HO-1 signaling pathways. DOX administration also induced the activation of NF-κB/MAPK signaling and increased the levels and expression of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) as well as expression of inflammatory mediators (iNOS and COX-2) in the heart. DOX also triggered DNA damage and apoptotic cell death in the myocardium. Additionally, histological studies revealed structural alterations of the myocardium. NERO treatment exhibited protection against the deleterious results of DOX on myocardium, as evidenced by the restoration of altered biochemical parameters, mitigated oxidative stress, inflammation, and apoptosis. The findings of the present study demonstrate that NERO provides cardioprotective effects against DOX-induced acute cardiotoxicity attributed to its potent antioxidant, anti-inflammatory, and antiapoptotic activities through modulating cellular signaling pathways.

Antioxidant and hepatoprotective property of squalene for counteracting the oxidative damage induced by methotrex- ate in experimental rats

Methotrexate (MTX), an antifolate drug, is extensively prescribed for patients suffering from diseases like cancer, psoriasis, neoplasms, and rheumatoid arthritis. Despite its effectiveness, MTX sometimes finds limited application because its undesirable side effects, including hepatic or renal impairment, bone marrow toxicity and gastrointestinal mucosal injury. Squalene, a highly unsaturated isoprenoid compound, isolated from shark liver oil has great potential in neutralizing the damaging effects triggered by free radicals. Therefore, in this study, the protective role of dietary squalene supplementation on oxidative stress induced by methotrexate in experimental rats was evaluated. A significant reduction was displayed in the activities of catalase (CAT) and superoxide dismutase (SOD) in MTX-intoxicated groups compared to other groups. Similarly, the activities of glutathione dependant enzymes (GPx and GST) and reduced glutathione (GSH) in MTX-induced groups were shown to be lower compared to the untreated control. Increased LPO (lipid peroxide) level was found in MTX-intoxicated groups compared to other groups. In addition, alterations in the levels of liver marker enzymes like AST, ALP, ALT, and LDH were noticed in MTX intoxicated groups compared to other groups. Biochemical results were confirmed by the histopathological examination of liver sections. In conclusion, the result obtained in the present study proposes that squalene exerts antioxidant activity and is capable of ameliorating oxidative stress and liver injury induced by MTX.

Columbianadin Dampens In Vitro Inflammatory Actions and Inhibits Liver Injury via Inhibition of NF-κB/MAPKs: Impacts on ∙OH Radicals and HO-1 Expression.

Columbianadin (CBN), a natural coumarin isolated from Angelica decursiva, is reported to have numerous biological activities, including anticancer and platelet aggregation inhibiting properties. Here, we investigated CBN’s anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell activation and deciphered the signaling process, which could be targeted by CBN as part of the mechanisms. Using a mouse model of LPS-induced acute liver inflammation, the CBN effects were examined by distinct histologic methods using trichrome, reticulin, and Weigert’s resorcin fuchsin staining. The result showed that CBN decreased LPS-induced expressions of TNF-α, IL-1β, and iNOS and NO production in RAW 264.7 cells and mouse liver. CBN inhibited LPS-induced ERK and JNK phosphorylation, increased IκBα levels, and inhibited NF-κB p65 phosphorylation and its nuclear translocation. Application of inhibitors for ERK (PD98059) and JNK (SP600125) abolished the LPS-induced effect on NF-κB p65 phosphorylation, which indicated that ERK and JNK signaling pathways were involved in CBN-mediated inhibition of NF-κB activation. Treatment with CBN decreased hydroxyl radical (•OH) generation and increased HO-1 expression in RAW 264.7 cells. Furthermore, LPS-induced liver injury, as indicated by elevated serum levels of liver marker enzymes (aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) and histopathological alterations, were reversed by CBN. This work demonstrates the utility of CBN against LPS-induced inflammation, liver injury, and oxidative stress by targeting JNK/ERK and NF-κB signaling pathways.

Curative Effects of Aqueous and Ethanol Stem Bark Extracts of Vitex doniana on Doxorubicin-Induced Cardiotoxicity in Rats

Background: Cardiovascular diseases (CVDs) constitute the number one cause of mortality at the global level, representing 30% of all global deaths. Therefore, finding ways to reduce deaths due to CVDs remain an important public health goal. Traditional healers in northern Nigeria use the stem bark of Vitex doniana to treat hypertensive patients. This study was aimed to investigate the cardiocurative potential of Vitex doniana on doxorubicin-induced Cardiotoxicity in rats.
 Methods: Thirty five (35) adult Albino rats weighing 175 ± 25 g were used, of which 30 were induced with cardiotoxicity by intraperitoneal injection of doxorubicin (10 mg/kg) for three consecutive days. Rats were treated by oral administration of Silymarin (100 mg/kg) and Vitex doniana aqueous or ethanol extract (100 mg/kg and 200 mg/kg) for 14 consecutive days and thereafter were sacrificed on the 15th day. Blood, plasma and serum were analyzed for lipid profile and serum markers for cardiotoxicity.
 Results: Phytochemical analysis of the extracts showed the presence of alkaloids, tannins, flavonoids, steroids, phenols, saponins, terpenoids and glycosides. Oral treatment with Vitex doniana extracts significantly (p<0.05) lowered the elevated levels of total cholesterol, triglycerides and LDL but significantly (p<0.05) increased the level of HDL (18.61 ± 0.55 mg/dl to 57.98 ± 0.78 mg/dl). The extracts also significantly (p<0.05) decreased the levels of serum marker enzymes for cardiotoxicity ALT, AST, CK – mb and LDH.
 Conclusion: The prophylactic cardiocurative use of Vitex doniana stem bark has been confirmed in this study as the extracts exhibited hypolipidemic and cardiocurative effects in dose dependent manner in doxorubicin-induced cardiotoxicity rat model.

Mast Cell Mediated Regulation of Small Intestinal Chloride Malabsorption in SAMP1/YitFc Mouse Model of Spontaneous Chronic Ileitis.

In Inflammatory Bowel Disease (IBD), malabsorption of electrolytes (NaCl) results in diarrhea. Inhibition of coupled NaCl absorption, mediated by the dual operation of Na:H and Cl:HCO3 exchangers on the brush border membrane (BBM) of the intestinal villus cells has been reported in IBD. In the SAMP1/YitFcs (SAMP1) mice model of spontaneous ileitis, representing Crohn’s disease, DRA (Downregulated in Adenoma) mediated Cl:HCO3 exchange was shown to be inhibited secondary to diminished affinity of the exchanger for Cl. However, NHE3 mediated Na:H exchange remained unaffected. Mast cells and their secreted mediators are known to be increased in the IBD mucosa and can affect intestinal electrolyte absorption. However, how mast cell mediators may regulate Cl:HCO3 exchange in SAMP1 mice is unknown. Therefore, the aim of this study was to determine the effect of mast cell mediators on the downregulation of DRA in SAMP1 mice. Mast cell numbers and their degranulation marker enzyme (β-hexosaminidase) levels were significantly increased in SAMP1 mice compared to control AKR mice. However, treatment of SAMP1 mice with a mast cell stabilizer, ketotifen, restored the β-hexosaminidase enzyme levels to normal in the intestine, demonstrating stabilization of mast cells by ketotifen. Moreover, downregulation of Cl:HCO3 exchange activity was restored in ketotifen treated SAMP1 mice. Kinetic studies showed that ketotifen restored the altered affinity of Cl:HCO3 exchange in SAMP1 mice villus cells thus reinstating its activity to normal. Further, RT-qPCR, Western blot and immunofluorescence studies showed that the expression levels of DRA mRNA and BBM protein, respectively remained unaltered in all experimental conditions, supporting the kinetic data. Thus, inhibition of Cl:HCO3 exchange resulting in chloride malabsorption leading to diarrhea in IBD is likely mediated by mast cell mediators.

Hepatoprotective Activity of Citrus Limetta on Wistar Albino Rats Hrishabh Jangid*, Mohammad Mukim, Rahul Ancheria, Ankit Sharma Kota College of Pharmacy, Kota, Rajasthan, India

The methanolic extract of citrus limetta (MECL) peel was screened for hepatoprotective activity in hepatotoxic wistar albino rats induced via paracetamol (PCM) and carbon tetra chloride (CCL4). Hepatoprotective activity of MECL was measured by estimating biochemical parameters such as AST (aspartate transaminase), ALT (alanine aminotransferase), SGPT (serum glutamine pyruvate transferase), SGOT (serum glutamic oxaloacetic transaminase). It was observed that administration of PCM, CCL4 decrease the level of proteins and increased the level of serum marker enzymes which is an evidence of existence of hepatotoxicity in rats. The present results provide strong evidence that MECL inhibits hepatotoxicity induced by CCL4 and PCM. The hepatoprotective action was much more significant at the dose of 2ml/kg when compared to 1mg/kg.

β-Caryophyllene, a natural bicyclic sesquiterpene attenuates β-adrenergic agonist-induced myocardial injury in a cannabinoid receptor-2 dependent and independent manner

The downregulation of cannabinoid type-2 receptors (CB2R) have been reported in numerous diseases including cardiovascular diseases (CVDs). The activation of CB2R has recently emerged as an important therapeutic target to mitigate myocardial injury. We examined whether CB2R activation can protect against isoproterenol (ISO)-induced myocardial injury (MI) in rats. In the present study, we investigated the cardioprotective effect of β-caryophyllene (BCP), a naturally occurring dietary cannabinoid in rat model of MI. Rats were pre- and co-treated with BCP (50 mg/kg, orally) twice daily for 10 days along with subcutaneous injection of ISO (85 mg/kg) at an interval of 24 h for two days (9th and 10th days). AM630 (1 mg/kg), a CB2 receptor antagonist, was injected intraperitoneal as a pharmacological challenge prior to BCP treatment to reveal CB2R-mediated cardioprotective mechanisms of BCP. Desensitization of beta-adrenergic receptor (β-AR) signaling, receptor phosphorylation and recruitment of adapter β-arrestins were observed in ISO-induced MI in rats. ISO injections caused impaired cardiac function, elevated the levels of serum cardiac marker enzymes, and enhanced oxidative stress markers along with altered PI3K/Akt and NrF2/Keap1/HO-1 signaling pathways. ISO also promoted lysosomal dysfunction along with activation of NLRP3 inflammasomes and TLR4-NFκB/MAPK signaling and triggered rise in proinflammatory cytokines. There was a concomitant mitochondrial dysfunction followed by the activation of endoplasmic reticulum (ER) stress-mediated Hippo signaling and intrinsic pathway of apoptosis as well as altered autophagic flux/mTOR signaling in ISO-induced MI. Furthermore, ISO also triggered dyslipidemia evidenced by altered lipids, lipoproteins and lipid marker enzymes along with ionic homeostasis malfunction. However, treatment with BCP resulted in significant protective effects on all biochemical and molecular parameters analyzed. The cardioprotective effects were further strengthened by preservation of cardiomyocytes and cell organelles as observed in histopathological and ultrastructural studies. Interestingly, treatment with AM630, a CB2R antagonist was observed to abrogate the protective effects of BCP on the biochemical and molecular parameters except hyperlipidemia and ionic homeostasis in ISO-induced MI in rats. The present study findings demonstrate that BCP possess the potential to protect myocardium against ISO-induced MI in a CB2-dependent and independent manner.

Molecular characterization and histo-physiological alterations induced by concurrent helminthosis in the liver of urban commensal rodents in Punjab, India.

Rodents harbour a number of parasites of public health importance, thus, they threaten human health and livestock. The present study aimed to characterize two helminthic species found in commensal rodents and record histo-physiological alterations induced by them. A total of 300 synanthropic rodents of three species: Rattus rattus (n=201), Bandicota bengalensis (n=90), and Mus musculus ( n=09 ) were live trapped and necropsied in different seasons during November 2017 to October 2019 at Ludhiana, Punjab, India. Liver of two species B. bengalensis (72.22%) and R. rattus (65.67%) were found infected with two helminthic parasites Taenia taeniaeformis, and Calodium hepaticum. These endoparasites were present either alone (4.33-6.33%) or as mixed infection (65.55%). The level of total proteins and liver marker enzymes including aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were found significantly higher in the liver of rodent species infected with single and mixed infection compared to those with no infection. In histopathological assay, granulomatous liver lesions and necrosis of hepatocytes were seen which were associated with eggs and adults of C. hepaticum and inflammatory reaction in hepatic parenchyma adjoining to cysts of T. taeniaeformis. Based upon scanning electron microscopy (SEM) identification and molecular characterization using mitochondrial cytochrome oxidase I (COI) region, the metacestodes in whitish cysts were confirmed to be of T. taeniaeformis for the first time in Punjab, India. The study highlights an alarmingly high infection of rodents with zoonotic parasites and suggests immediate pest (rodent) control to check the dissemination of zoonotic diseases by helminth species under study.

Synergistic effect of ursolic acid and piperine in CCl4 induced hepatotoxicity

Background Ursolic acid (UA) is a potent plant-based hepatoprotective agent having poor bioavailability, which hampers its therapeutic efficacy. The present study tries to overcome this limitation by combining it with piperine (PIP), a proven bioenhancer and hepatoprotective agent. Methods The type of interaction (synergism, addition, or antagonism) resulting between UA and PIP was analyzed and quantified by isobologram and combination index analysis. The hepatoprotective activity of UA and PIP was evaluated by measuring the level of hepatic marker enzymes. Pharmacokinetic analysis was carried out to ascertain the improvement of bioavailability. Results The combinations significantly decrease the enzyme levels, which indicate better hepatoprotective activity compared to single drugs. The relative oral bioavailability of UA was increased about tenfold (from AUC0– t =12.78 ± 2.59 µg/h/ml to 125.15 ± 1.84 µg/h/ml) along with the improvement of plasma concentration and elimination half-life. Conclusions The findings indicated that the combination of PIP and UA is an effective strategy in enhancing the bioavailability and hepatoprotective potential of UA. KEY MESSAGES Ursolic acid in a combination with piperine provides a synergistic hepatoprotective effect in carbon tetrachloride induced liver damage in rats. Piperine improves the pharmacokinetic properties of ursolic acid when given in combination. Piperine improves the relative oral bioavailability of ursolic acid by tenfold when combined together.

Anticancer Effect of Troxerutin in Human Non-Small-Cell Lung Cancer Cell A549 and Inhibition of Tumor Formation in BALB/c Nude Mice.

This study is intended to explore the anticancer, antiproliferative, and chemopreventive action of troxerutin (TX) in human non-small-cell lung cancer cell (A549) using BALB/c nude mice. 2 × 106 A549 cells were subcutaneously injected into mice, along with 10 μM and 20 μM/kg body weight of TX orally for 19 days. On the last day, tumor weight and volume were assessed. Stress marker enzymes such as Aryl hydrocarbon hydroxylase (AHH), lactate dehydrogenase (LDH), 5'Nucleotidase (5'ND), and γ-glutamyltranspeptidase (γ-GT) were estimated in the lung tissues. Cytotoxicity of TX was assessed using MTT assay. Expression of carcinoembryonic antigen (CEA) and inflammatory cytokines were also analyzed. Histopathological examination of tissue sections and immunohistochemical examination of proliferating cell nuclear antigen (PCNA) were also performed. mRNA expression of p53, p21, cyclin D1, P13k, Akt, and mTOR were analyzed using RT-PCR. TX administered orally in a dose-dependent manner markedly reverted the level of stress marker enzymes to a significant extent. TX also exhibited significant protection against lung cancer cells, as evidenced by cytotoxicity assay and histopathological studies. It was also found to reduce the expression of PCNA, cyclin D1, P13k, Akt, and mTOR, but increase the expression of p53 and p21. TX has also been shown to reduce cancer cell inflammation, as was evidenced by reduced expression of inflammatory cytokines. Thus TX could be used as an effective chemopreventive and anticancer agent in treating cancer.

Hepatoprotective Potential of Endophytic Aspergillus Niger Strain A6 Fractions Isolated from Phyllanthus Amarus Schum and Thonn

Aim To isolate characterize and screen for the in vitro antioxidant and hepatoprotective effects of P1EA and P1nB extracts from the endophytic fungi Aspergillus niger strain A6 PALF-1 isolated from leaves of Phyllanthus amarus in paracetamol induced hepatotoxicity.Materials and methods PALF-1 was isolated from Phyllanthus amarus leaves by standard procedures. Chloroform P1C ethyl acetate P1EA and n-butanol P1nB extracts were drawn up. P1C P1EA and P1nB were screened for in vitro antioxidant activity against 2 2-diphenyl-1-picrylhydrazyl DPPH hydroxyl radical followed by reducing power assay. P1EA 50 and 100 mgkg b. w. and P1nB 50 and 100 mgkg b. w. were screened for hepatoprotective activity against paracetamol model in rats. High performance thin layer chromatography HPTLC profile of P1EA and P1nB was also studied. Characterization of PALF-1 was done by polymerase chain reaction PCR sequential analysis.Results P1EA and P1nB found to scavenge DPPH hydroxyl radicals and also showed reductive ability. In animal studies administration of paracetamol 2 gmkg b. w. significantly increased the levels of liver biochemical marker enzymes in comparison to normal group. A marked decrease in the levels of superoxide dismutase SOD and catalase CAT was also observed. Administration of P1EA and P1nB 50 amp100 mg kg significantly reduced the actions of paracetamol by lowering the levels of serum biochemical parameters at various significance levels plt0.001 plt0.01 increasing the levels of SOD and CAT plt0.001 and decreased lipid peroxidation LPO. rDNA PCR sequencing analysis phylogenetic and molecular studies of PALF-1 confirmed the endophytic fungus as Aspergillus niger strain A6. HPTLC finger printing of P1EA and PnB showed the presence of polyvalent compounds.Conclusion P1EA and P1nB fractions of Aspergillus niger strain A6 showed significant antioxidant and hepatoprotective activity in paracetamol induced hepatotoxicity. This may be attributed to the antioxidant principles present in P1EA and P1nB which need to be explored.

Naringin, a natural flavonone glycoside attenuates N-nitrosodiethylamine- induced hepatocellular carcinoma in sprague-dawley rats

Background: Hepatocellular Carcinoma (HCC) being proclaimed as the world's fifth common cancer shows a high mortality rate due to delayed diagnosis. Regular day-to-day activities expose us to promotive factors that potentiate further to risk of liver damage, cirrhosis, and carcinogenesis. Matrix metalloproteinases (MMPs), a proteolytic enzyme, involved in cancer invasion are considered as a prognostic biomarker for HCC. Naringin (NAR), a flavanone glycoside, is recognized to have therapeutic efficacy in HCC. Objectives: The present research work focuses on evaluation of chemotherapeutic efficiency, NAR against HCC in Sprague–Dawley rats. Materials and Methods: In vitro studies were primarily carried out to estimate the cell viability of NAR in HepG2 cells followed by further justifications through in vivo studies in N-nitrosodiethylamine developed HCC. The efficacy of NAR at doses 30mg/kg/day and 60mg/kg/day was assessed through estimation of liver marker enzymes, antioxidant levels, glycoproteins, and level of MMP-2 and 9 to confirm the hypothesis. Results: While in vitro results revealed pronounced potency of NAR in inhibiting HepG2 cell viability, NAR concurrently stabilized the serum levels of liver marker enzymes, antioxidant levels, and serum glycoproteins. It also significantly reduced the levels of MMPs in treatment groups which was confirmed by zymography. Conclusion: With the support of these results, it can be concluded that NAR attenuates HCC by controlling the alterations in morphological, biochemical, and histopathological parameters that make it a suitable candidate as a potential chemotherapeutic agent against HCC.

Quercetin with lycopene modulates enzymic antioxidant genes pathway in isoproterenol cardiotoxicity in rats

ABSTRACT Quercetin (QN) is a naturally occurring phenolic compound found largely in vegetables and fruits. Lycopene (LY) is yet another natural phytocompound, found abundantly in red-colored fruits and vegetables. Both have been reported to have beneficial activities in humans. In this study, we document in vivo experimental model for isoproterenol (ISO) cardiac injury toxicity in Sprague-Dawley (SD) rats and treatment with a combined optimized concentration of quercetin and lycopene (QL). Male SD rats of different groups were treated with QL (80 mg/kg QN and 3 mg/kg LY together p.o.) for 10 days with ISO administration (100 mg/kg i.p.) on days 7 and 8. After experimental period, CK-MB, TROP, AST, ALT, LDH, GST, GPx, CAT, SOD, Vit.E, Vit. C, GSH, GSSG and MDA were estimated. SD rats administered with ISO showed an obvious rise in the serum marker enzyme levels and tissue oxidative stress markers (MDA and GSSG). Furthermore, marked reductions in the body weight and increases enzymic and non-enzymic antioxidant levels were noticed. Histological features of the heart also indicated a disruption in the cardiac myofibrils structure of ISO-intoxicated rats. Also, quantitative PCR analysis revealed an involvement of antioxidant and related pathway genes such as Nrf2, HO-1, NQO1, GSTµ, SOD1, SOD2, CAT and BCl-2 genes. QL pretreatment prevented all these adverse effects of ISO cardiotoxicity and significantly reduced the myocardial damage. Decrease in oxidative stress was observed, possibly through alterations in the expression levels of enzymic antioxidant genes (GSTµ, SOD1, SOD2 and CAT). In general, QL exert a strong protective effect through the modulations in enzymic antioxidant activity and associated molecular pathways-regulating effect in cardiovascular disease.

Dietary Black Seed Effects on Growth Performance, Proximate Composition, Antioxidant and Histo-Biochemical Parameters of a Culturable Fish, Rohu (Labeo rohita)

This feeding trial was conducted to investigate the effects of dietary black seed (Nigella sativa) supplementation on the growth performance, muscles proximate composition, antioxidant and histo-biochemical parameters of rohu (Labeo rohita). Fingerlings (8.503 ± 0.009 g) were fed on 0.0%, 1% and 2.5% black seed supplemented diets for 28 days. Fish sampling was done on the 7th, 14th, 21st and 28th day of experiment. The results of the present study indicated that black seed supplementation significantly increased growth performance and muscles protein contents of rohu over un-supplemented ones. Lipid peroxidation levels significantly decreased in all the studied tissues (liver, gills, kidney and brain) of black seed fed rohu, whereas the antioxidant enzymes (catalase, glutathione-S-transferase, glutathione peroxidase and reduced glutathione) activities were increased in all the studied tissues of black seed supplemented rohu at each sampling day. The hepatic-nephric marker enzymes levels were decreased for black seed fed rohu. The present study showed that tested black seed levels are safe for rohu. Black seed is cheaply available in local markets of Pakistan; therefore, based on the results of the present study, it is suggested that black seed has potential to be used as natural growth promoter and antioxidant in the diet of rohu.

Kuanxiong aerosol inhibits apoptosis and attenuates isoproterenol-induced myocardial injury through the mitogen-activated protein kinase pathway

Ethnopharmacological relevanceKuanxiong aerosol (KXA) is a common clinical drug based on Fangxiang Wentong (FXWT) therapy in the treatment of angina pectoris. However, the pharmacological mechanism of KXA in the prevention and treatment of myocardial injury (MI) is not clear. Aim of the studyThe purpose of this study was to explore the protective effect of KXA on isoproterenol (ISO)-induced MI in rats. Materials and methodsThe study included male Wistar Kyoto rats (age: 6 weeks). The rats were randomly divided into the following 5 groups (n = 6 per group): control group, ISO group, isosorbide mononitrate (ISMN) group (5 mg/kg), KXA-L group (0.1 mL/kg), and KXA-H group (0.3 mL/kg). The rats in the last three groups were given intragastric administration for 14 days, and rats in control group and ISO group were given the same amount of normal saline daily. ISO (120 mg/kg) was used to induce MI on the 13th and 14th days. We assessed electrocardiograms (ECGs), myocardial specific enzymes, histopathological changes, and apoptosis. ResultsWe found that KXA reduced the increase in the ST-segment amplitude (elevation or depression) and the levels of myocardial marker enzymes induced by ISO in MI rats, improved the pathological changes in myocardial tissue, and reduced cardiomyocyte apoptosis. At the same time, KXA significantly inhibited the up-regulation of caspase-3 and Bax expression and down-regulation of Bcl-2 expression induced by ISO. RNA sequencing showed that 90 up-regulated genes induced by ISO were down-regulated after KXA treatment, whereas 27 down-regulated genes induced by ISO were up-regulated after KXA treatment. In addition, KEGG pathway enrichment analysis showed that the mitogen-activated protein kinase (MAPK) signaling pathway may be an important target of KXA in the treatment of ISO-induced MI in rats. The results of RNA sequencing verified by Western blot analysis showed that KXA significantly inhibited the activation of the ISO-induced MAPK pathway. ConclusionsKXA improves cardiac function in MI rats by inhibiting apoptosis mediated by the MAPK signaling pathway.

Antioxidant, anti-cancer and ameliorative activities of Spirulina platensis and pomegranate juice against hepatic damage induced by CCl4

Due to the excessive impact of synthetic drugs, unravelling and employing safe, natural alternatives are now needed to resolve a number of diseases. In this research, we have evaluated hepatoprotective and antioxidant activities of Spirulina platensis and pomegranate juice in rats against hepatotoxicity induced by carbon tetrachloride (CCl4). Spirulina crude carotenoid extract was screened by UPLC-MS / MS. Activities of liver marker enzymes; measured aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and determined lipid peroxidation and antioxidant status as reduced glutathione (GSH) in liver homogenate. The infusion of CCl4 (2 ml/kg b.wt) greatly increases levels of liver marker enzymes and lipid peroxidation, resulting in depletion of antioxidants. Treatment of Spirulina platensis (Sp), pomegranate juice (Pj)or mixture (PJSP) of Spirulina water extract 10% and pomegranate juice 90% (1 ml/100 g b.wt) to CCl4-disrupted rats resulted in decreased activity of liver marker enzymes, lipid peroxidation with increased antioxidant status. Chromatographic separation showed that ß-carotene is the predominant carotenoid extract. This carotenoid extract was tested for colon carcinoma (HCT-116), liver carcinoma (HepG2) and intestinal carcinoma cell lines (CACO) LC50 for 21.8, 14 and 11.3 ug / ml, respectively. Total phenolic phytochemicals, total carotenoids and total flavonoids were also measured in Spirulina. Our study clearly demonstrates that Spirulina platensis and pomegranate juice had hepatoprotective effect on CCl4-caused hepatotoxicity in rats through its antioxidant activity.

Optimized piperine–phospholipid complex with enhanced bioavailability and hepatoprotective activity

Piper species is one of the most widely consumed spices for culinary purposes. Piperine (PIP) present in Piper species has a wide range of therapeutic activity including hepatoprotection. However, the major biological limitation of PIP is its low bioavailability after oral administration. Purpose of the study was to prepare an optimized and adequately characterized PIP–phospholipid complex (PPC) as a delivery system to overcome these limitations and to investigate the pharmacokinetics and hepato-protectivity of the formulation in the animal model. Response surface methodology was adopted to optimize the process parameters for PPC preparation. FT-IR, DTA, PXRD, SEM, molecular docking etc. were used for characterization. Solubility, log P, dissolution efficiency and in vivo pharmacokinetics were also investigated. PPC showed enhanced hepatoprotective potential as compared to pure PIP at the same dose level (25 and 50 mg/kg). PPC restored the levels of serum marker and antioxidant enzymes. PPC also increased the bioavailability of PIP in rat serum by 10.40-fold in comparison with pure PIP at the same dose level and enhanced the elimination half-life (t1/2 el) from 0.477 ± 1.76 to 9.80 ± 1.98 h. Results concluded that PPC enhanced the hepatoprotection of PIP which may be due to the improved bioavailability and pharmacokinetics of PIP in rats.