Curdione Plays an Important Role in the Inhibitory Effect ofCurcuma aromaticaon CYP3A4 in Caco-2 Cells

Curcuma aromaticais a plant belonging to genusCurcumaof familyZingiberaceaeand is widely used as supplements in Japan. Rhizomes ofC. aromaticahave curcumin as a major yellow pigment and curdione as a main ingredient of essential oils. In this study, we investigated the affect ofC. aromaticaon CYP3A4 using 1α,25-(OH)2-D3-treated Caco-2 clone cells. Caco-2 cells were treated with methanol extract (0.1 mg ml−1), its hexane soluble fraction (0.1 mg ml−1), curcumin (4 μM) and curdione (20 μM) for 72 hours. Nifedipine was used as a substrate of CYP3A4. Methanol extract, hexane fraction and curdione inhibited the formation of oxidized nifedipine by 50–70%, and curcumin showed no effect. The IC50s of methanol extract, hexane fraction and curdione to oxidized nifedipine formation were 21, 14 and 3.9 μg ml−1(16.9 μM), respectively. The content of curdione in methanol extract was 11.4%. Moreover, all of methanol extract, hexane fraction and curdione decreased CYP3A4 protein expression but had no affect on CYP3A4 mRNA expression. Our results showed that these drugs further decreased the CYP3A4 protein expression level after the protein synthesis was inhibited by cychroheximide. These findings suggest that curdione plays an important role in the CYP3A4 inhibitory activity ofC. aromaticaand curdione might inhibit the activity by accelerating the degradation of CYP3A4.

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Long non-coding RNA RP11-284P20.2 promotes cell proliferation and invasion in hepatocellular carcinoma by recruiting EIF3b to induce c-met protein synthesis

Bioscience Reports ◽

10.1042/bsr20200297 ◽

2020 ◽

Vol 40 (3) ◽

Cited By ~ 1

Author(s):

Qin-Liang Fang ◽

Jian-Yin Zhou ◽

Yu Xiong ◽

Cheng-Rong Xie ◽

Fu-Qiang Wang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Protein Synthesis ◽

Protein Expression ◽

Mrna Expression ◽

Translation Initiation Factor ◽

Expression Level ◽

Mrna Expression Level ◽

Protein Expression Level ◽

Proliferation And Invasion

Abstract A newly identified lncRNA designated as RP11-284P20.2 has been identified to be up-regulated in hepatocellular carcinoma (HCC), but its role in HCC remain poorly understood. Quantitative PCR and immunocytochemical analysis were performed using the HCC tissues to identify the potential interaction partners of RP11-284P20.2. Moreover, RP11-284P20.2 was knocked down in HCC cell lines, HepG2 and SMMC7721, to investigate the influence of this lncRNA on cell growth properties. Additionally, RNA fluorescence in situ hybridization and immunofluorescence, RNA immunoprecipitation, and RNA pull-down assays were performed to determine the interaction of RP11-284P20.2 with c-met mRNA and eukaryotic translation initiation factor 3b (EIF3b). Silencing RP11-284P20.2 inhibited cell viability, migration, invasion, and colony formation, and increased apoptosis. Overexpression of c-met abolished these effects of RP11-284P20.2 in HCC cells. Histopathological examination showed that HCC tissues with high RP11-284P20.2 expression had higher c-met protein level than that in HCC tissues with low RP11-284P20.2 expression. However, there was no positive correlation between the expression levels of RP11-284P20.2 and c-met mRNA. RP11-284P20.2 knockdown led to a decease in c-met protein expression level, but did not affect the c-met mRNA expression level. These data suggest that RP11-284P20.2 regulates c-met protein expression level, which is independent of c-Met mRNA expression level. It was also confirmed that RP11-284P20.2 has high affinity toward both c-met mRNA and EIF3b protein, and hence RP11-284P20.2 probably recruits EIF3b protein to c-met mRNA and further facilitates its translation. RP11-284P20.2 promotes cell proliferation and invasion in hepatocellular carcinoma by recruiting EIF3b to induce c-met protein synthesis.

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Comprehensive Analysis of ESRRA in Endometrial Cancer

Technology in Cancer Research & Treatment ◽

10.1177/1533033821992083 ◽

2021 ◽

Vol 20 ◽

pp. 153303382199208

Author(s):

Shufang Wang ◽

Xinlong Huo

Keyword(s):

Endometrial Cancer ◽

Protein Expression ◽

Immune Cell ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

The Cancer Genome Atlas ◽

Expression Level ◽

Operating Characteristics ◽

Protein Expression Level ◽

Normal Tissues

Background: Estrogen-related receptor alpha (ESRRA) was reported to play an important role in multiple biological processes of neoplastic diseases. The roles of ESRRA in endometrial cancer have not been fully investigated yet. Methods: Expression data and clinicopathological data of patients with uteri corpus endometrial carcinoma (UCEC) were obtained from The Cancer Genome Atlas (TCGA). Comprehensive bioinformatics analysis was performed, including receiver operating characteristics (ROC) curve analysis, Kaplan-Meier survival analysis, gene ontology (GO) enrichment analysis, and Gene Set Enrichment Analysis (GSEA). Immunohistochemistry was used to detect the protein expression level of ESRRA and CCK-8 assay was performed to evaluate the effect of ESRRA on the proliferation ability. Results: A total of 552 UCEC tissues and 35 normal tissues were obtained from the TCGA database. The mRNA and protein expression level of ESRRA was highly elevated in UCEC compared with normal tissues, and was closely associated with poor prognosis. ROC analysis indicated a very high diagnostic value of ESRRA for patients with UCEC. GO and GSEA functional analysis showed that ESRRA might be mainly involved in cellular metabolism processes, in turn, tumorigenesis and progression of UCEC. Knockdown of ESRRA inhibited the proliferation of UCEC cells in vitro. Further immune cell infiltration demonstrated that ESRRA enhanced the infiltration level of neutrophil cell and reduced that of T cell (CD4+ naïve), NK cell, and cancer associated fibroblast (CAF). The alteration of immune microenvironment will greatly help in developing immune checkpoint therapy for UCEC. Conclusions: Our study comprehensively analyzed the expression level, clinical value, and possible mechanisms of action of ESRRA in UCEC. These findings showed that ESRRA might be a potential diagnostic and therapeutic target.

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Lipopolysaccharide-induced sensitization of adenylyl cyclase activity in murine macrophages

AJP Cell Physiology ◽

10.1152/ajpcell.00171.2005 ◽

2006 ◽

Vol 290 (1) ◽

pp. C143-C151 ◽

Cited By ~ 20

Author(s):

Y. Osawa ◽

H. T. Lee ◽

C. A. Hirshman ◽

D. Xu ◽

C. W. Emala

Keyword(s):

Protein Synthesis ◽

Adenylyl Cyclase ◽

Actinomycin D ◽

Inhibitory Effect ◽

Cytokine Release ◽

Adenylyl Cyclase Activity ◽

Phosphorylation State ◽

Murine Macrophages ◽

Dependent Pathway ◽

New Protein

LPS is known to modulate macrophage responses during sepsis, including cytokine release, phagocytosis, and proliferation. Although agents that elevate cAMP reverse LPS-induced macrophage functions, whether LPS itself modulates cAMP and whether LPS-induced decreases in proliferation are modulated via a cAMP-dependent pathway are not known. Murine macrophages (RAW264.7 cells) were treated with LPS in the presence or absence of inhibitors of prostaglandin signaling, protein kinases, CaM, Giproteins, and NF-κB translocation or transcription/translation. LPS effects on CaMKII phosphorylation and the expression of relevant adenylyl cyclase (AC) isoforms were measured. LPS caused a significant dose (5–10,000 ng/ml)- and time (1–8 h)-dependent increase in forskolin-stimulated AC activity that was abrogated by pretreatment with SN50 (an NF-κB inhibitor), actinomycin D, or cycloheximide, indicating that the effect is mediated via NF-κB-dependent transcription and new protein synthesis. Furthermore, LPS decreased the phosphorylation state of CaMKII, and pretreatment with a CaM antagonist attenuated the LPS-induced sensitization of AC. LPS, cAMP, or PKA activation each independently decreased macrophage proliferation. However, inhibition of NF-κB had no effect on LPS-induced decreased proliferation, indicating that LPS-induced decreased macrophage proliferation can proceed via PKA-independent signaling pathways. Taken together, these findings indicate that LPS induces sensitization of AC activity by augmenting the stimulatory effect of CaM and attenuating the inhibitory effect of CaMKII on isoforms of AC that are CaMK sensitive.

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Identification of Secondary Metabolites and Antibacterial Activity of Non Polar Fraction from Heterotrigona itama Propolis

Journal of Fundamental and Applied Pharmaceutical Science ◽

10.18196/jfaps.v2i1.12406 ◽

2021 ◽

Vol 2 (1) ◽

pp. 23-33

Author(s):

Abdul Aziz ◽

Veggy Nadya Yuliawan ◽

Paula Mariana Kustiawan

Keyword(s):

Antibacterial Activity ◽

Methanol Extract ◽

Phenolic Content ◽

Polar Fraction ◽

Hexane Fraction ◽

Diffusion Method ◽

Total Phenolic ◽

Phytochemical Analysis ◽

Phytochemical Content ◽

Color Changes

Propolis is one of the natural products produced by kelulut bees and is still not widely used. The type of stingless bee that is the prima donna in the community is Heterotrigona itama. This study aims to determine the phytochemical content of the n-hexane fraction of Heterotrigona itama bee propolis collected from Kutai Kartanegara, East Kalimantan. The n-hexane fraction was obtained from the methanol extract of H. itama propolis by the liquid-liquid partition method. After obtaining the n-hexane fraction, the research continued with a qualitative phytochemical test to identify the compound and determine total phenolic. Antibacterial activity was determined by the agar well diffusion method with a serial concentration in Escherichia coli bacteria. Qualitative phytochemical analysis in the form of color changes showed that the n-hexane fraction of H. itama propolis contained flavonoids, alkaloids, saponins, and tannins. Based on the results, the total phenolic content of the n-hexane fraction sample was 490 mgGAE/100 g. It caused the n-hexane fraction to have lower phenolic content than the methanol extract (792 mg GAE100 g). Furthermore, this result indicated that the non-polar fraction was not substantial enough to extracted phenolic compounds. It correlated to the antibacterial activity of the n-hexane fraction, which was very weak (2 mm ± 1.5) at 200µg/mL concentration.

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The silencing of long non-coding RNA ANRIL suppresses invasion, and promotes apoptosis of retinoblastoma cells through the ATM-E2F1 signaling pathway

Bioscience Reports ◽

10.1042/bsr20180558 ◽

2018 ◽

Vol 38 (6) ◽

Cited By ~ 8

Author(s):

Yang Yang ◽

Xiao-Wei Peng

Keyword(s):

Protein Expression ◽

Signaling Pathway ◽

Inhibitory Effect ◽

Down Regulation ◽

Reading Frame ◽

Non Coding Rna ◽

Retinoblastoma Cells ◽

Human Retinoblastoma ◽

Y79 Cells ◽

Long Non Coding Rna

As one of the most common primary intraocular carcinomas, retinoblastoma generally stems from the inactivation of the retinoblastoma RB1 gene in retinal cells. Antisense non-coding RNA in the INK4 locus (ANRIL), a long non-coding RNA (lncRNA), has been reported to affect tumorigenesis and progression of various cancers, including gastric cancer and non-small cell lung cancer. However, limited investigations emphasized the role of ANRIL in human retinoblastoma. Hence, the current study was intended to investigate the effects of ANRIL on the proliferation, apoptosis, and invasion of retinoblastoma HXO-RB44 and Y79 cells. The lentivirus-based packaging system was designed to aid the up-regulation of ANRIL and ATM expressions or employed for the down-regulation of ANRIL in human retinoblastoma cells. Afterward, ANRIL expression, mRNA and protein expression of ATM and E2F1, and protein expression of INK4b, INK4a, alternate reading frame (ARF), p53 and retinoblastoma protein (pRB) were determined in order to elucidate the regulation effect associated with ANRIL on the ATM-E2F1 signaling pathway. In addition, cell viability, apoptosis, and invasion were detected accordingly. The results indicated that the down-regulation of ANRIL or up-regulation of ATM led to an increase in the expressions of ATM, E2F1, INK4b, INK4a, ARF, p53, and pRB. The silencing of ANRIL or up-regulation of ATM exerted an inhibitory effect on the proliferation and invasion while improving the apoptosis of HXO-RB44 and Y79 cells. In conclusion, the key observations of our study demonstrated that ANRIL depletion could act to suppress retinoblastoma progression by activating the ATM-E2F1 signaling pathway. These results provide a potentially promising basis for the targetted intervention treatment of human retinoblastoma.

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Development of mouthwash with Rosmarinus officinalis extract

Brazilian Journal of Pharmaceutical Sciences ◽

10.1590/s1984-82502014000400020 ◽

2014 ◽

Vol 50 (4) ◽

pp. 851-858 ◽

Cited By ~ 1

Author(s):

Isabela Moreira Baumgratz de Paula ◽

Flávia Costa Moraes ◽

Orlando Vieira de Souza ◽

Célia Hitomi Yamamoto

Keyword(s):

Sodium Fluoride ◽

Inhibitory Activity ◽

Raw Materials ◽

Ethanol Extract ◽

Inhibitory Effect ◽

Rosmarinus Officinalis ◽

Hexane Fraction ◽

Diffusion Method ◽

Oral Infections

Rosmarinus officinalis, which belongs to the Lamiaceaefamily, is a species of medicinal flora with therapeutic properties. In order to exploit the benefits of these properties, a mouthwash formulation was developed, with careful selection of raw materials to meet pharmacotechnical requirements. Extracts of the plant were incorporated into a mouthwash, which was shown to have inhibitory action in vitro against the micro-organisms commonly found in periodontics. Controls for assessing the quality of the drugs were carried out, quantifying phenols and flavonoids as chemical markers. Mouthwash solutions were formulated containing 0.1, 5 and 10% ethanol extract of R. officinalis; and 0.05, 5 and 10% of the hexane fraction of R. officinalis. In order to evaluate synergism, ethanol extract and hexane fraction were also added to formulations containing 0.05% sodium fluoride and 0.12% chlorhexidine digluconate. These formulations were assessed for inhibitory effect against the specific microorganisms involved in the process of bacterial plaque formation, S. mutans(ATCC25175) and C. albicans(ATCC 10231), frequently found in cases of oral infections. The agar diffusion method was used to evaluate the inhibitory activity of extracts and formulations. All mouthwash solutions displayed inhibitory activity having higher sensitivity to S. mutansfor the 5% ethanol extract+0.05% sodium fluoride, and greater sensitivity to C. albicansfor the 10% hexane fraction. Results were characterized by the appearance of a growth inhibition halo, justifying the utilization and association of extracts of R. officinalis.

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Remedial Hypoglycemic Activity of n-Hexane Fraction of Hydro-Methanol Extract of Seed ofSwietenia mahagoni(L.) jacq. in Streptozotocin-Induced Diabetic Rat: A Comparative Evaluation

Journal of Herbs Spices & Medicinal Plants ◽

10.1080/10496475.2014.895920 ◽

2014 ◽

Vol 21 (1) ◽

pp. 38-58 ◽

Cited By ~ 1

Author(s):

Tushar Kanti Bera ◽

Kausik Chatterjee ◽

Debidas Ghosh

Keyword(s):

Comparative Evaluation ◽

Methanol Extract ◽

Hypoglycemic Activity ◽

Hexane Fraction ◽

Diabetic Rat

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The radical scavenging activity of 2-2’ diphenyl -1- picrylhydrazil (DPPH) on the methanol extracts and Ethyl acetate fractions of red dragon fruit peel (Hylocereus polyrhizus (F.A.C.Weber) Britton dan Rose)

International Journal of Phytomedicine ◽

10.5138/09750185.1945 ◽

2017 ◽

Vol 9 (1) ◽

pp. 79

Author(s):

Sri Wahdaningsih ◽

Subagus Wahyuono ◽

Sugeng Riyanto ◽

Retno Murwanti

Keyword(s):

Antioxidant Activity ◽

Ethyl Acetate ◽

Methanol Extract ◽

Soluble Fraction ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Insoluble Fraction ◽

Fruit Peel ◽

Activity Test ◽

Dragon Fruit

Red dragon fruit (H. Polyrhizus) is one of the the plants that has a great potential as natural antioxidant. This study tested the activity of radical scavenging of 2-2' diphenyl -1- pikril hidrazil (DPPH) in the methanol extract, as well as in the soluble and insoluble fractions of ethyl acetate of red dragon fruit peel. This research is carried out through various stages, such as: extraction and fractionation to obtain both insoluble fraction and soluble fractions of ethyl acetate. Antioxidant activity test is conducted by the method of thin layer chromatography and spectrophotometry. Antioxidant activity test, IC50 values of methanol extract, ethyl acetate soluble fraction, and insoluble fraction of ethyl acetate had been obtained consecutively as much as 241.19 µg /mL, 8.34 µg/mL, 46.84 µg/mL. The soluble fraction of ethyl acetate had greater antioxidant activity compared to the methanol extract and the insoluble fractions of ethyl acetate.

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