Methanolic extract of Woodfordia fruticosa Kurz flowers ameliorates carbon tetrachloride-induced chronic hepatic fibrosis in rats

2014 ◽  
Vol 32 (7) ◽  
pp. 1224-1236 ◽  
Author(s):  
A Nitha ◽  
SP Prabha ◽  
PN Ansil ◽  
MS Latha
Keyword(s):  
Carbon Tetrachloride ◽  
Hepatic Fibrosis ◽  
Methanolic Extract ◽  
Mass Spectrometry Analysis ◽  
Dependent Manner ◽  
Spectrometry Analysis ◽  
Collagen Iii ◽  
Male Wistar Rats ◽  
Matrix Accumulation ◽  
Woodfordia Fruticosa

Hepatic fibrosis, characterized by extracellular matrix accumulation, is the common cause of chronic liver failure and is a leading cause of morbidity and mortality worldwide. The aim of the present study was to evaluate the effect of dried flowers of Woodfordia fruticosa on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rat model. Hepatic fibrosis was induced in male Wistar rats by CCl4 administration (150 μl/100 g rat weight, oral) twice a week for 10 weeks. In preventive model, administration of daily doses of methanolic extract of W. fruticosa (MEWF) at two different doses (100 mg/kg, body weight (b.w.) and 200 mg/kg, b.w.) was started 1 week before the onset of CCl4 administration and continued for 10 weeks. In curative model, MEWF at 100 and 200 mg/kg were given for last 2 weeks after the establishment of fibrosis. MEWF at a dose of 200 mg/kg was able to exert a more pronounced effect as evidenced histologically by significant reduction in fibrotic septa formation in liver tissue, immunohistochemically by abridged expression of collagen III, and also biochemically by serum and tissue antioxidant status, lipid peroxidation, and hydroxyproline level. Liquid chromatography–mass spectrometry analysis revealed the presence of confertin, quercetin methyl ether, ellagic acid, and stigmasterol in MEWF, which could be responsible for its antifibrotic activity. These results indicate the effective protection exerted by MEWF against CCl4-induced hepatic fibrosis in a dose-dependent manner.

scholarly journals ENIGMATIC INDUCTION OF CYTOMIXIS IN ALLIUM CEPA ROOT MERISTEM BY AGLAIA EDULIS ROXB. LEAF EXTRACT AND ITS PHYTOCHEMICAL RATIONALE

2020 ◽  
pp. 168-171
Author(s):  
ARCHANA ELAMKULAM RAVINDRAN ◽  
JOHN ERNEST THOPPIL
Keyword(s):  
Allium Cepa ◽  
Leaf Extract ◽  
Root Meristem ◽  
Methanolic Extract ◽  
Synergistic Action ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Phytochemical Analysis ◽  
Spectrometry Analysis ◽  
Dose Dependent

Objective: The present study aims to analyze the potential of Aglaia edulis Roxb. leaf extract to induce cytological aberrations in Allium cepa root meristem and to determine the phytoconstituents in the extract. Methods: Cytotoxicity evaluation of the leaf methanolic extract was done using Allium cepa assay using various concentrations. Volatile phytoconstituents in the extract were determined using gas chromatography–mass spectrometry analysis. Results: Considerable number of cytomictic cells along with other aberrations was observed. The occurrence of cytomixis was found to be dose dependent where it ranged from 6.58±0.35 to 29.45±0.45. The percentage of cytomictic cells among the total aberrant cells was observed between 35.19±1.67 and 77.39±1.39. The phytochemical analysis of the plant extract revealed the presence of active secondary metabolites. Conclusion: The synergistic action of the active compounds might have triggered the phenomenon of cytomixis which, in turn, could be exploited for the production of polyploids.

Arginine Methylation Of RBM15 By PRMT1 Controls Alternative Splicing Of Genes Involved In Megakaryocyte Differentiation

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2443-2443
Author(s):  
Xinyang Zhao ◽  
Li Zhang ◽  
Rui Wang ◽  
Ngoc Tung Trans ◽  
Hairui Su ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Rna Binding ◽  
Conflicts Of Interest ◽  
Chromosome Translocation ◽  
Arginine Methylation ◽  
Mass Spectrometry Analysis ◽  
Dependent Manner ◽  
Hematopoietic Stem ◽  
Spectrometry Analysis ◽  
Megakaryocyte Differentiation

Abstract More than 90% of under one year old infants with acute megakaryoblastic leukemia (AMKL) have chromosome translocation t(1;22)(p13;q13) with RBM15 fused to MKL1. RBM15 encodes an RNA binding protein important for hematopoietic stem cell self-renewal and differentiation. In agreement with its roles in AMKL, RBM15 is required for normal megakaryocyte differentiation. We found that higher expression of PRMT1 (Protein Arginine Methyltransferase) is commonly seen in M7 leukemia patient samples than other types of myeloid leukemia and that RBM15 is a bona fide methylation target for PRMT1. Using mass spectrometry analysis, we mapped the PRMT1 mediated mono-methylated site. The enzymatic activity of the PRMT1 V2 isoform is required for RBM15 degradation, as both shRNA molecules knocking down PRMT1 and small chemical PRMT1 inhibitors stabilize the RBM15 protein. Mutation of the methylation site to lysine blocks the ubiquitylation mediated degradation. Thus the degradation is a methylation dependent process. We identified the E3 ligase responsible for the degradation. Down-regulation of the RBM15 protein changes the isoform ratio of genes including GATA1 critical megakaryocyte differentiation. We found that RBM15 regulates its interaction with SF3B1A in methylation dependent manner during alternative splicing of GATA1 pre-mRNA. Thus, via methylation triggered RBM15 degradation, the megakaryocyte progenitor cells maintain a delicate balance between differentiation and proliferation by keeping the proper ratio of GATA1s and GATA1-full length mRNA. SF3B1A has been shown to be mutated in myeloid dysplasia syndrome and in several different types of leukemia. Methylation by PRMT1 links the two types of leukemic genes into a single pathway. Our results imply that targeting PRMT1/RBM15 pathway might be a potential therapy for AMKL and other blood malignancies. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Antiproliferative Activities of Methanolic Extract and Fractions of Tetrapleura Tetraptera Fruit

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Anastasia Rosebud Aikins ◽  
Peggy Afua Birikorang ◽  
Mary Chama ◽  
Eunice Dotse ◽  
Abigail Anning ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Methanolic Extract ◽  
Colorimetric Assay ◽  
Jurkat Cells ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Cancer Chemotherapeutics ◽  
Tetrapleura Tetraptera ◽  
Antiproliferative Activities ◽  
Mcf 7

Most of the current cancer chemotherapeutics are associated with harsh and undesirable side effects, including toxicity and chemoresistance, driving the need for safer and more effective alternatives. In this study, the antiproliferative activities of the methanolic extract of Tetrapleura tetraptera fruits and nine different fractions (C1–C9) from the column chromatographic separation of the extract against leukemia (Jurkat) and human breast cancer (MCF-7) cell lines were investigated using a tetrazolium-based colorimetric assay. Phytochemical screening of the extract and fractions found alkaloids, carbohydrates, flavonoids, glycosides, phenols, saponins, steroids, tannins, and terpenoids in the methanolic extract. Most of the fractions exhibited antiproliferative activity (>100 μg/mL) with the Jurkat cells being more susceptible than the MCF-7 cells. Four of the collected fractions C4, C3, C5, and C2 had good selective indices in decreasing order of activity, in the case of Jurkat cells. Liquid chromatography-mass spectrometry analysis of all samples (except for C4 and C9) revealed that C1, C2, C3, and C5 each had a single component. More importantly, fractions C2, C3, and C5, which were selective to Jurkat cells, also had the same retention time of 1.846 min. Fractions C6 and C8 had two components, with C7 having four components. This study serves as a basis for further work to isolate and characterize potential anticancer agents from the fractions of extracts of T. tetraptera fruits.

Hypoxia-driven secretion of extracellular matrix proteins in the exosomes reflects the asymptomatic pathology of rotator cuff tendinopathies

2020 ◽  
Author(s):  
Finosh G. Thankam ◽  
Devendra K. Agrawal
Keyword(s):  
Extracellular Matrix ◽  
Rotator Cuff ◽  
Molecular Pathology ◽  
Matrix Proteins ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Tendon Cells ◽  
Collagen Iii ◽  
Asymptomatic Phase ◽  
Expression Status

The major hallmark of rotator cuff tendinopathies (RCT) is the disorganization of tendon extracellular matrix (ECM) which is due to decrease in the collagen I-to-collagen III ratio. In addition, the pathology of tendon matrisome remains asymptomatic and hypoxia has been identified to be the priming signal to initiate the molecular pathology of RCT. Also, the secretome content of hypoxia challenged tendon cells (tenocytes) reflects the pathological status of RCT. With this background, the present study was designed to establish the expression status and the molecular crosstalk of the ECM component proteins contained in the exosomes of the hypoxia challenged swine tenocytes. The mass spectrometry analysis revealed the upregulation of COL1A2, P4HA1, PRDX2, P3H1, COL6A1, PPIB, LCN1, and COL3A1 and the downregulation of COLA12, PDIA4, COLG, FN1, CTSK, and TNC in the exosomes of hypoxic tenocytes. These proteins interact with diverse proteins and operate multiple pathways associated with ECM homeostasis and repair as determined by NetworkAnalyst. The functional analysis of these proteins reflects the pathology of tendon ECM which is correlated with the asymptomatic phase of RCT. Understanding the signaling mediated by these proteins would reveal the underlying molecular pathology and offers translational significance in the diagnosis/management of RCT.

Anti-Fibrotic Effects of Neferine on Carbon Tetrachloride-Induced Hepatic Fibrosis in Mice

2015 ◽  
Vol 43 (02) ◽  
pp. 231-240 ◽  
Author(s):  
Mo-Si Chen ◽  
Jia-Hua Zhang ◽  
Jia-Ling Wang ◽  
Lu Gao ◽  
Xiao-Xu Chen ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Hepatic Fibrosis ◽  
Liver Tissue ◽  
Intraperitoneal Injection ◽  
Dependent Manner ◽  
Model Group ◽  
Liver Index ◽  
Seed Embryo ◽  
Dose Dependent ◽  
Tgf Β1

The effects of neferine, a bisbenzylisoquinline alkaloid extracted from the seed embryo of the Chinese traditional medicine Nelumbo nucifera Gaertn, on carbon tetrachloride ( CCl 4)-induced hepatic fibrosis in mice were evaluated. Adult male Kunming mice were administered with CCl 4 1 ml/kg via intraperitoneal injection twice a week for 8 weeks. At the beginning of the 9th week, mice were treated with normal saline, colchicine (0.1 mg/kg), and neferine (5, 10, 20 mg/kg) via intraperitoneal injection once a day for 2 weeks. The liver index and histological examination, plasma ALT/AST levels, hydroxyproline and TGF-β1 content of liver tissue were examined. In the model group, the liver index, the hydroxyproline content of liver tissue and plasma ALT/AST levels were increased, and a high expression of TGF-β1 was observed. The abnormal changes could be improved by neferine in a dose-dependent manner. Our data showed that neferine had an antifibrosis effect on CCl 4-induced hepatic fibrosis in mice, possibly partly due to the decreased expression of TGF-β1 in the liver.

scholarly journals Efficacy of the Aqueous Extract of Azadirachta indica Against the Marine Parasitic Leech and Its Phytochemical Profiling

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 1908
Author(s):  
Balu Alagar Venmathi Maran ◽  
Dawglas Josmeh ◽  
Jen Kit Tan ◽  
Yoong Soon Yong ◽  
Muhammad Dawood Shah
Keyword(s):  
Mass Spectrometry ◽  
Aqueous Extract ◽  
Azadirachta Indica ◽  
Total Mortality ◽  
Mass Spectrometry Analysis ◽  
Economic Losses ◽  
Dependent Manner ◽  
Spectrometry Analysis ◽  
Orbitrap Mass Spectrometry ◽  
Q Exactive

Zeylanicobdella arugamensis (Hirudinea), a marine parasitic leech, not only resulted in the mortality of the host fish (Groupers) but also caused economic losses. The current study aimed to elucidate the antiparasitic efficacy of the aqueous extract of the Azadirachta indica leaves against Z. arugamensis and to profile the composition via LC-Q Exactive HF Orbitrap mass spectrometry. Different concentrations (25, 50 and 100 mg/mL) of A. indica extract were prepared and tested on the parasitic leeches. The total mortality of leeches was noticed with an exposure to the A. indica aqueous extract. The average times required for the aqueous extract at concentrations of 25, 50 and 100 mg/mL to kill the leeches were 42.65 ± 9.20, 11.69 ± 1.11 and 6.45 ± 0.45 min, respectively, in a dose-dependent manner. The Orbitrap mass spectrometry analysis indicated the presence of five flavonoids (myricetin 3-O-galactoside, trifolin, isorhamnetin, quercetin and kaempferol), four aromatics (4-methoxy benzaldehyde, scopoletin, indole-3-acrylic acid and 2,4-quinolinediol), three phenolics (p-coumaric acid, ferulic acid and phloretin) and two terpenoids (pulegone and caryophyllene oxide). Thus, our study indicates that A. indica aqueous extract is a good source of metabolites with the potential to act as a biocontrol agent against the marine parasitic leech in aquaculture.

scholarly journals YTHDF2 alleviates cardiac hypertrophy via regulating Myh7 mRNA decoy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hongfei Xu ◽  
Zhen Wang ◽  
Miao Chen ◽  
Wenting Zhao ◽  
Tingting Tao ◽  
...  
Keyword(s):  
Cardiac Hypertrophy ◽  
Mass Spectrometry Analysis ◽  
Immunofluorescent Staining ◽  
Dependent Manner ◽  
Protective Effects ◽  
Spectrometry Analysis ◽  
Pathological Cardiac Hypertrophy ◽  
Therapeutic Mechanisms ◽  
M6a Rna Methylation ◽  
Underlying Mechanisms

Abstract Background Pathological cardiac hypertrophy is a major contributor of heart failure (HF), which seriously threatens human’s health world widely. Deregulation of m6A RNA methylation, and m6A methyltransferases and de-methyltransferases have been demonstrated to act essential roles in cardiac hypertrophy and HF. Here, we studied the potential roles and its underlying mechanisms of m6A Reader YTHDF proteins in HF. In this study, we constructed HF mouse model by transverse aortic constriction surgery. Primary cardiomyocytes were isolated and stimulated with isoproterenol (ISO) or phenylephrine (PHE) to induce myocardial hypertrophy. Results Through single-cell RNA-seq analysis, immunofluorescent staining, HE staining, Western blotting, and real time-PCR detections, we found that YTHDF2 mRNA and protein level, but not YTHDF1 or YTHDF3, was significantly increased during HF development. YTHDF2 overexpression could efficiently alleviate cardiac hypertrophy. Furthermore, through immunoprecipitation accompanied with mass spectrometry analysis, Gene Ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, we found that ISO stimulation did not evidently affect YTHDF2-interacting proteins. However, ISO or PHE stimulation significantly increased YTHDF2 protein interacting with Myh7 (beta-myosin heavy chain) mRNA, an important cardiac hypertrophy marker, in an m6A-dependent manner. Knockdown of Myh7 or deletion of the YTH domain of YTHDF2 reversed the protective effects of YTHDF2 on cardiac hypertrophy. Finally, we found that ISO or PHE stimulation promoted YTHDF2 protein expression through enhancing Ythdf2 mRNA stability in an m6A-dependent manner in cardiomyocytes. Conclusions Overall, our results indicate that the m6A Reader YTHDF2 suppresses cardiac hypertrophy via Myh7 mRNA decoy in an m6A-dependent manner. This study highlights the functional importance of YTHDF2-dependent cardiac m6A mRNA regulation during cardiac hypertrophy, and provides a novel mechanistic insight into the therapeutic mechanisms of YTHDF2.

scholarly journals Soluble Fas ligand drives autoantibody-induced arthritis by binding to DR5/TRAIL-R2

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Dongjin Jeong ◽  
Hye Sung Kim ◽  
Hye Young Kim ◽  
Min Jueng Kang ◽  
Hyeryeon Jung ◽  
...  
Keyword(s):  
Fas Ligand ◽  
Affinity Purification ◽  
Mass Spectrometry Analysis ◽  
Dependent Manner ◽  
Chemokine Production ◽  
Spectrometry Analysis ◽  
Soluble Fas ◽  
Soluble Fas Ligand ◽  
Deficient Mice

To date, no study has demonstrated that soluble Fas ligand (sFasL)-mediated inflammation is regulated via interaction with Fas in vivo. We found that FasL interacts specifically with tumor necrosis factor receptor superfamily (TNFRSF)10B, also known as death receptor (DR)5. Autoantibody-induced arthritis (AIA) was attenuated in FasL (Faslgld/gld)- and soluble FasL (FaslΔs/Δs)-deficient mice, but not in Fas (Faslpr/lpr and Fas–/–)- or membrane FasL (FaslΔm/Δm)-deficient mice, suggesting sFasL promotes inflammation by binding to a Fas-independent receptor. Affinity purification mass spectrometry analysis using human (h) fibroblast-like synovial cells (FLSCs) identified DR5 as one of several proteins that could be the elusive Fas-independent FasL receptor. Subsequent cellular and biochemical analyses revealed that DR5 interacted specifically with recombinant FasL–Fc protein, although the strength of this interaction was approximately 60-fold lower than the affinity between TRAIL and DR5. A microarray assay using joint tissues from mice with arthritis implied that the chemokine CX3CL1 may play an important downstream role of the interaction. The interaction enhanced Cx3cl1 transcription and increased sCX3CL1 production in FLSCs, possibly in an NF-κB-dependent manner. Moreover, the sFasL–DR5 interaction-mediated CX3CL1–CX3CR1 axis initiated and amplified inflammation by enhancing inflammatory cell influx and aggravating inflammation via secondary chemokine production. Blockade of FasL or CX3CR1 attenuated AIA. Therefore, the sFasL–DR5 interaction promotes inflammation and is a potential therapeutic target.

The functional significance of ATM phosphorylation of Bub3 on Serine 135 in sensitivity to DNA damaging agents.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e15048-e15048
Author(s):  
Siyue Zhang ◽  
Mingming Xiao ◽  
Zhuang Liu ◽  
Yaqi Mo ◽  
Hong Liu ◽  
...  
Keyword(s):  
Dna Damage ◽  
Survival Rates ◽  
Mitotic Checkpoint ◽  
Immunofluorescence Assay ◽  
Mass Spectrometry Analysis ◽  
Dependent Manner ◽  
Atm Kinase ◽  
Spectrometry Analysis ◽  
Dna Damaging Agents ◽  
Mitotic Checkpoint Protein

e15048 Background: Identification of biomarkers to assess and modify the sensitivity of cancer cells to radiotherapy and chemotherapy is critical to improve cancer treatment outcome. Budding uninhibited by benzimidazoles 3 (Bub3) is a mitotic checkpoint protein, and it is frequently overexpressed in many cancers and associated with low survival rates. Bub3 is involved in the repair of DNA damage induced by radiotherapy and chemotherapy. We recently identified that the ATM kinase phosphorylated Bub3 on Serine 135 (S135) via the stable isotope labeled amino acid in cell culture -mass spectrometry analysis. This study aims to explore the mechanism of ATM-phosphorylated Bub3 in the DNA damage response (DDR) and its effect on tumor sensitivity to DNA damaging agents. Methods: The radiosensitivity of the cells was detected by clonal formation assay, the proliferation ability of the cells was detected by the MTS assay. The expression of DDR protein γ-H2AX in the nucleus was detected by immunofluorescence assay. Genomic instability was observed by multinuclear formation. Co-immunoprecipitation and Western Blot were used to explore the internal mechanism of ATM phosphorylation of Bub3 in DDR. Results: We showed that ionizing radiation (IR) could induce Bub3 S/TQ (the specific ATM consensus motif) phosphorylation in an ATM dependent manner. Mutation of Bub3 Serine 135 to alanine (S135A) led to a phosphorylation defect. Phenotypic experiments showed hypersensitivity to IR in cells expressing Bub3 S135A. Bub3 S135A prolonged existence of γ-H2AX foci and increased the proportion of cells containing micronuclei. Further, we found that Ku70 and Ku80 showed a significant increase after IR in their interactions with Bub3, while Bub3 S135A mutation significantly reduced the interaction, leading to impaired DNA repair. Conclusions: We demonstrate that Bub3 S135 phosphorylation mediated by ATM is essential for an optimal DDR and disruption of this pathway increases tumor sensitivity to DNA damaging agents.

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