Anti-inflammatory activity of Alpinia officinarum hance on rat colon inflammation and tissue damage in DSS induced acute and chronic colitis models

The purpose of this study was to investigate the possible prophylactic effects of Alpinia officinarum hance on experimentally induced acute and chronic colitis models,in-vivo and in-vitro.Acute and chronic colitis were induced in Male Wistar rats by administration of Dextran Sulfate Sodium(DSS)in drinking water.DSS induction exhibited colon shrinkage,increased the Disease Activity Index(DAI)score,increased the levels of inflammatory markers and caused severe anemia.DSS induced animals,co-treated with the hexane extract of Alpinia officinarum(HEAO)(200 mg/kg body wt),effectively suppressed colonic injury that was evidenced by the reduced DAI score,colon weight/length ratio,histological damage,proinflammatory markers and MPO activity.Further,it restored the colonic antioxidants near to normal levels by regulating the oxidative stress via attenuation of lipid peroxidation.Our results revealed that the degree of colitis caused by the administration of DSS was significantly attenuated by HEAO.In addition,the in-vitro study showed that HEAO treatment inhibited the proliferation of HT-29 cells and down regulated the mRNA expression of NF-B and COX-2.Taken together,these results suggest that HEAO is a promising anti-oxidant and anti-inflammatory agent that support its possible therapeutic role in the treatment of colitis.

Potential antioxidant and cytoprotective effects of essential oil extracted from Cymbopogon citratus on OxLDL and H_(2)O_(2) LDL induced Human Peripheral Blood Mononuclear Cells(PBMC)

Cymbopogon citratus(lemon grass)is commonly used in traditional folk medicine.The essential oil extracted from C.citratus has been reported as a potential anti-oxidant and anti-inflammatory agent.This study has been designed to explore the protective effect of C.citratus(lemon grass)against modified LDL(OxLDL and H2O2 LDL)induced cytotoxicity in Peripheral Blood Mononuclear Cells(PBMC).The essential oil extracted from C.citratus(EOC)was subjected to FT-IR spectroscopic analysis for the identification of functional groups.In vitro antioxidant assays were carried out to assess the electron donating capability of EOC as compared with a known standard L-ascorbic acid.The cytoprotective effects of EOC were determined in PBMC induced with modified LDL.Spectra obtained from FT-IR analysis showed the presence of functional groups in EOC such as H-bonded,O-H stretching,N-H stretching,aldehyde-C-H stretching,aldehyde/ketone-C=O stretching,-C=C-stretching,-CH_(3) bending,-C-H in plane bending.EOC has greater antioxidant property when compared with the standard L-ascorbic acid.EOC at all test concentrations demonstrated free radical scavenging activity and cytoprotective effect when challenged against modified LDL in PBMC.The above results show EOC as a promising antioxidant and cytoprotective agent.

Defensive nature of Sargassum polycystum (Brown alga) against acetaminophen-induced toxic hepatitis in rats: Role of drug metabolizing microsomal enzyme system, tumor necrosis factor-a and fate of liver cell structural integrity

AIM： To assess the defensive nature of Sargassum polycystum （S. polycystum） （Brown alga） against acetaminophen （AAP）-induced changes in drug metabolizing microsomal enzyme system, tumor necrosis factor （TNF-α） and fine structural features of the liver during toxic hepatitis in rats. METHODS： Male albino Wistar strain rats used for the study were randomly categorized into 4 groups. Group Ⅰ consisted of normal control rats fed with standard diet. Group II rats were administered with acetaminophen （800 mg/kg body weight, intraperitoneally）. Group Ⅲ rats were pre-treated with S. polycystum extract alone. Group IV rats were orally pre-treated with S. polycystum extract （200 mg/kg body weight for 21 d） prior to acetaminophen induction （800 mg/kg body weight, intraperitoneally）. Serum separated and liver was excised and microsomal fraction was isolated for assaying cytochrome P450, NADPH Cyt P450 reductase and bs. Serum TNF-α was detected using ELISA. Fine structural features of liver were examined by transmission electron microscopy. RESULTS： Rats intoxicated with acetaminophen showed considerable impairment in the activities of drug metabolizing microsomal enzymes, such as cytochrome P450, NADPH Cyt P450 reductase and bs when compared with the control rats. The rats intoxicated with acetaminophen also significantly triggered serum TNF-a when compared with the control rats. These severe alterations in the drug metabolizing enzymes were appreciably pretreated with S. polycystum. prevented in the rats The rats pretreated withS. polycystum showed considerable inhibition in the elevation of TNF-α compared to the rats intoxicated with acetaminophen. The electron microscopic observation showed considerable loss of structural integrity of the endoplasmic reticulum, lipid infiltration and ballooning of mitochondria in the acetaminophen-intoxicated rats, whereas the rats treated with S. polycystum showed considerable protection against acetaminophen-induced alterations in structural integrity. CONCLUSION： These observations suggest that the animals treated with S. polycystum extract may have the ability to protect the drug metabolizing enzyme system and mitochondrial functional status from free radical attack, thereby showing its defense mechanism in protecting hepatic cells from acetaminophen toxic metabolite N-acetyl-para-benzoquinone-imine （NAPQI）.

Ferroptosis regulates key signaling pathways in gastrointestinal tumors:Underlying mechanisms and therapeutic strategies

Ferroptosis is an emerging novel form of non-apoptotic,regulated cell death that is heavily dependent on iron and characterized by rupture in plasma membrane.Ferroptosis is distinct from other regulated cell death modalities at the biochemical,morphological,and molecular levels.The ferroptotic signature includes high membrane density,cytoplasmic swelling,condensed mitochondrial membrane,and outer mitochondrial rupture with associated features of accumulation of reactive oxygen species and lipid peroxidation.The selenoenzyme glutathione peroxidase 4,a key regulator of ferroptosis,greatly reduces the lipid overload and protects the cell membrane against oxidative damage.Ferroptosis exerts a momentous role in regulating cancer signaling pathways and serves as a therapeutic target in cancers.Dysregulated ferroptosis orchestrates gastrointestinal(GI)cancer signaling pathways leading to GI tumors such as colonic cancer,pancreatic cancer,and hepatocellular carcinoma.Crosstalk exists between ferroptosis and other cell death modalities.While apoptosis and autophagy play a detrimental role in tumor progression,depending upon the factors associated with tumor microenvironment,ferroptosis plays a decisive role in either promoting tumor growth or suppressing it.Several transcription factors,such as TP53,activating transcription factors 3 and 4,are involved in influencing ferroptosis.Importantly,several molecular mediators of ferroptosis,such as p53,nuclear factor erythroid 2–related factor 2/heme oxygenase-1,hypoxia inducible factor 1,and sirtuins,coordinate with ferroptosis in GI cancers.In this review,we elaborated on key molecular mechanisms of ferroptosis and the signaling pathways that connect ferroptosis to GI tumors.

Thymoquinone inhibits the migration of mouse neuroblastoma(Neuro-2a) cells by down-regulating MMP-2 and MMP-9

Thymoquinone(TQ), an active component derived from the medial plant Nigella sativa, has been used for medical purposes for more than 2 000 years. Recent studies have reported that TQ blocked angiogenesis in animal model and reduced migration, adhesion, and invasion of glioblastoma cells. We have recently shown that TQ could exhibit a potent cytotoxic effect and induce apoptosis in mouse neuroblastoma(Neuro-2a) cells. In the present study, TQ treatment markedly decreased the adhesion and migration of Neuro-2a cells. TQ down-regulated MMP-2 and MMP-9 protein expression and m RNA levels and their activities. Furthermore, TQ significantly down-regulated the protein expression of transcription factor NF-κB(p65) but not significantly altered the expression of N-Myc. Taken together, our data indicated that TQ's inhibitory effect on the migration of Neuro-2a cells was mediated through the suppression of MMP-2 and MMP-9 expression, suggesting that TQ treatment can be a promising therapeutic strategy for human malignant neuroblastoma.

Lycopene stabilizes lipoprotein levels during D-galactosamine/lipopolysaccharide induced hepatitis in experimental rats

Objective:To investigate the effect of lycopene on lipoprotein metabolism during D-galactosamine/lipopolysacchoride(D-Gal/LPS)induced hepatitis in experimentul rats.Methods:The efficacy of lycopene was validated during D-Gal/LPS induced hepatitis by analyzing the activity of lipid metabolizing enzymes such as lipoprotein lipase(LPL),lecithincholesterol acyl transferase(LCAT)and hepatic triglyceride lipase(HTCL).Lipo protein analyses were done by the estimation of very low density lipoprotein cholesterol(VLDL),low density lipoprotein cholesterol(LDL)and high density lipoprotein cholesterol(HDL).Remits:The toxic insult of D-galactosamine/lipopolysaccharide(D-Gal/LPS)in experimental group of animals reduces the normal values of lipid metabolizing enzymes due to liver injury.The significant drop in the levels of HDL and concomitant increase in the values of VLDL and LDL were observed.The pretreatment of lycopene restore these altered values to near normal level in experimental group of animals.Conclusions:In the light of results,it can be concluded that administration lycopene stabilizes the lipoprotein levels by regulating the lipid metabolizing enzymes through its antioxidant defense and helps to maintain the normal lipid metabolism during toxic injury in liver.

Hesperetin conjugated PEGylated gold nanoparticles exploring the potential role in anti-inflammation and anti-proliferation during diethylnitrosamine-induced hepatocarcinogenesis in rats

Liver cancer is the fifth most common cancer and one of the leading causes of death in the world, and second most common cause of death in men. Natural products emerge as the most enduring approaches in the development of anticancer targeting drug. Hesperetin(HP), one of the abundant flavonoids found naturally in citrus fruits, has received considerable attention in anti-cancer promotion and progression. The present study was conducted to decipher the role of 0.5 ml hesperetin conjugated gold nanoparticles(Au-m PEG(5000)-S-HP NPs) during diethylnitrosamine(DEN)-induced hepatocarcinogenesis in male Wistar albino rats and shows the better antioxidant that possesses anti-inflammatory,anti-proliferation and anticarcinogenic properties and may modulate signaling pathways.The confirmation of polymer functionalized gold nanoparticles and drug loaded polymer gold nanoparticles were characterized by HR-TEM with EDAX, and DLS with Zeta potential techniques. The drug encapsulation efficiency and release properties were carried out in PBS at pH 7.4 for Au-mPEG(5000)-S-HP and compared with the control pure hesperetin(HP).Here, we review the role of mast cell counts, tumor necrosis factor alpha(TNF-α), transcription factor nuclear factor-κB(NF-κB), levels of glycoconjugates, proliferating cell nuclear antigen(PCNA) and argyrophilic nucleolar organizing regions, are the master regulator of inflammation and proliferation, in the development of hepatocellular injury, liver fibrosis and HCC. DEN-administered animals showed increased mast cell counts, tumor necrosis factor alpha, transcription factor nuclear factor-κB, glycoconjugates, proliferating cell nuclear antigen, and argyrophilic nucleolar organizing regions. Whereas Au-mPEG(5000)-S-HP NPs supplementation considerably suppressed all the above abnormalities. These results suggest that the Au-m PEG(5000)-S-HP NPs exhibited the better potential anticancer activity by inhibiting cell inflammation and proliferation in DEN-induced hepatocellular carcinogenesis.

Asiaticoside, a trisaccaride triterpene induces biochemical and molecular variations in brain of mice with parkinsonism

Background:Parkinson’s disease characterized by oxidative stress and mitochondrial damage in the pars compacta of substantia nigra remains a challenge to manage with an added disadvantage of side effects of L-levo dopa,the standard drug used for therapy.Thus,an alternative approach of utilizing natural components would be beneficial in the management of the disease.The present study was aimed to investigate the potential role of asiaticoside(As),a trisaccaride triterpene against1–methyl 4–phenyl 1,2,3,6 tetrahydropyridine(MPTP)-induced neurotoxicity in experimental mice.Methods:Mice were divided into 4 groups:Group I received vehicle saline,group II was treated with 20 mg/kg of body weight of MPTP(2 doses with 2 h intervals),group III received MPTP along with 50 mg/kg body weight of As for the 21 consecutive days starting from the day of MPTP intoxication.Group IV received 50 mg/kg body weight of asiaticoside for the same period serving as drug control.Animals were sacrificed at the end of experimental period and the striatum and midbrain samples were analyzed for enzyme assays,transmission electron microscopic(TEM)analysis.Immunofluorescent assay was performed to study the expression of GFAP to detect astrocyte,which are activated due to neuronal damage.Imunohistochemical studies were carried out to quantify the expression of Bax and Bcl2,the molecular signatures that would provide clues of the extent of neurodegeneration.Results:The activities of enzymes were increased on As administration when compared with those of group II animals.Expressions of Bax and Bcl2 along with GFAP did show significant variations(p<0.05)on MPTP treatment when compared to control animals and the changes were found to be reversed significantly(p<0.05)after treatment with asiaticoside.TEM analysis also showed attenuated degenerative architecture on As administration.The mice which received As alone(drug control IV)did not show significant variation from that of the control mice.Conclusion:The observations suggest that asiaticoside may be efficacious in protecting neurons from the oxidative damage caused by the insult of MPTP.

Acute and 28-day repeated oral toxicological evaluation of Kuruthi Azhal Chooranam-a Siddha preparation on rodents

Objective:To determine the effect of phytochemicals in acute and repeated dose of 28-day oral toxicity of Kuruthi Azhal Chooranam(KAC)in Sprague Dawley rats of both sexes.Methods:Acute oral toxicity was conducted with 2000 mg/kg body weight of KAC orally and the treated animals were observed for signs of toxicity at 30 min,1,2,4 and 24 h and for up to 14 days.In repeated 28-day oral toxicity study,the KAC formulation was administered orally with 600,900 and 1200 mg/kg body weight/day to all the three groups of rats.The animals were observed for clinical signs of toxicity,mortality and morbidity throughout the study.Also body weight,feed consumption,haematological,plasma biochemistry and serum electrolytes,gross pathology,weights of the organ and histology were studied for no-observed-adverse-effect level.High dose of KAC formulation and control reversal groups were also included for delayed toxic effects determination.Results:In the acute toxicity study of KAC formulation,2000 mg/kg body weight dose exhibited no toxic signs and mortality during study.In sub-acute 28-day repeated dose toxicity study,there was no significant difference found between control and KAC treated groups(body weight,haematology,biochemistry and serum electrolytes).No abnormalities was found in gross pathology,organs weight and histological observation after KAC treatment.Conclusions:The current study suggests that LD_(50)of KAC was>2000 mg/kg and no-observed-adverse-effect level was>1200 mg/kg/day in rats.KAC could be used as Siddha drug for various indications.

Carvacrol Promotes Cell Cycle Arrest and Apoptosis through PI3K/AKT Signaling Pathway in MCF-7 Breast Cancer Cells

Objective:To examine the role of carvacrol in modulating PI3 K/AKT signaling involved in human breast cancer pathogenesis using in vitro experimental model MCF-7 cells.Methods:MTT and lactate dehydrogenase assays were performed with cells treated with different doses of carvacrol(0–250μmol/L)at different time points(24 and 48 h).The nuclear morphology was assessed in MCF-7 cells with propidium iodide(PI)and acridine orange/ethidium bromide(AO/EB)staining and analyzed by fluorescence microscopy.Events like cell cycle arrest and apoptosis were observed by flow cytometric analysis and expressions of p-Rb,cyclin D1,cyclin-dependent kinase 4(CDK4),CDK6,Bax,Bcl-2,PI3 K/p-AKT were analyzed by immunoblot.Results:Carvacrol significantly reduced cell viability with the half maximal inhibitory concentration value of 200μmol/L at 24 and 48 h(P<0.05).importantly,there was a significant increase in the accumulation of the G0/G1 phase upon treatment with carvacrol in MCF-7 cells(P<0.05 or P<0.01).A remarkable decrease in protein expressions of p-Rb,cyclin D1,CDK4 and CDK6 denoted cell cycle arrest(P<0.05 or P<0.01).In addition,carvacrol treatment significantly inhibited PI3 K/p-AKT protein expressions leading to induction of apoptosis mediated by decreased Bcl2 and increased Bax protein expressions.Further,Annexin V/PI staining by FACS analysis,dual staining by AO/EB and PI staining studies suggested induction of apoptosis by carvacrol through PI3 K/Akt signaling pathway in MCF-7 cells.Conclusion:Carvacrol significantly inhibited the breast cancer MCF-7 cell proliferation and induced apoptosis via suppressing PI3/AKT signaling pathway.