Molecular Mechanism of Hepatitis C Virus-Induced Glucose Metabolic Disorders

Hepatitis C virus (HCV) infection causes not only intrahepatic diseases but also extrahepatic manifestations, including metabolic disorders. Chronic HCV infection is often associated with type 2 diabetes. However, the precise mechanism underlying this association is still unclear. Glucose is transported into hepatocytes via glucose transporter 2 (GLUT2). Hepatocytes play a crucial role in maintaining plasma glucose homeostasis via the gluconeogenic and glycolytic pathways. We have been investigating the molecular mechanism of HCV-related type 2 diabetes using HCV RNA replicon cells and HCV J6/JFH1 system. We found that HCV replication down-regulates cell surface expression of GLUT2 at the transcriptional level. We also found that HCV infection promotes hepatic gluconeogenesis in HCV J6/JFH1-infected Huh-7.5 cells. HCV infection transcriptionally up-regulated the genes for phosphoenolpyruvate carboxykinase (PEPCK) and glucose 6-phosphatase (G6Pase), the rate-limiting enzymes for hepatic gluconeogenesis. Gene expression of PEPCK and G6Pase was regulated by the transcription factor forkhead box O1 (FoxO1) in HCV-infected cells. Phosphorylation of FoxO1 at Ser319 was markedly diminished in HCV-infected cells, resulting in increased nuclear accumulation of FoxO1. HCV NS5A protein was directly linked with the FoxO1-dependent increased gluconeogenesis. This paper will discuss the current model of HCV-induced glucose metabolic disorders

Promising Anti-Hepatitis C Virus Compounds from Natural Reseurcees

Hepatitis C virus (HCV) infection is a major worldwide problem, which involves approximately 170 million people. High morbidity of patients is caused by chronic infection, which leads to liver cirrhosis, hepatocellular carcinoma and other HCV-related diseases. The sustained virological response (SVR) has been markedly improved to be >90% by the current standard interferon (IFN)-free treatment regimens with a combination of direct-acting antiviral agents (DAAs) targeting the viral NS3 protease, NS5A multi-function protein and NS5B RNA-dependent RNA polymerase, compared with 50–70% of SVR rates achieved by the previous standard IFN-based treatment regimens with or without an NS3 protease inhibitor. However, the emergence of DAA-resistant HCV strains and the limited access to the DAAs due to their high cost could be major concerns. Also, the long-term prognosis of patients treated with DAAs, such as the possible development of hepatocellular carcinoma, still needs to be further evaluated. Natural resources are considered to be good candidates to develop anti-HCV agents. Here, we summarize anti-HCV compounds obtained from natural resources, including medicinal plant extracts, their isolated compounds and some of their derivatives that possess high antiviral potency against HC

Aktivitas Sitotoksisitas Ekstrak Metanol Daun Sirsak (Annona muricata L.) terhadap Karsinoma Hepatoseluler Strain HUH7IT-1 Cell Line

ABSTRAKLatar Belakang: Karsinoma hepatoseluler (HCC) merupakan tumor ganas hati primer dengan prognosis pada umumnya dapat menyebabkan kematian. Studi awal penelitian antiviral hepatitis C pada tumbuhan Sirsak (Annona muricata L.) pada konsentrasi 20 μg/mL memperlihatkan toksisitas yang sangat tinggi terhadap Huh7it-1 cell line, yang diindikasi memiliki potensi anti kanker terhadap sel hati, sehingga penelitian ini bertujuan menguji beberapa konsentrasi lebih rendah pada ekstrak metanol daun Annona muricata L. (EMDAM) terhadap Karsinoma Hepatoseluler strain Huh7it-1 cell line.Metode: Sel diuji dengan konsentrasi 20, 10, 5, 2.5, 1.25, 0.6, 0.3 μg/mL selama 48 jam. Sitotoksisitas EMDAM terhadap Huh7it-1 dilihat dengan mikrokop inverted dan selanjutnya diukur dengan metode MTT [3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium].Hasil: Hasil uji menunjukkan sel memperlihatkan bentuk tidak monolayer pada mikroskop inverted dengan sitotoksisitas hingga konsentrasi terendah pada 0.3 μg/mL mencapai 84,7%, sehingga konsentrasi 50% Sitotoksisitas (CC50) < 0.3 μg/mL.Simpulan: Hasil uji mengindikasi bahwa EMDAM memiliki potential terhadap aktivitas anti kanker hati. Studi lebih lanjut diperlukan untuk purifikasi untuk senyawa aktif sebagai antikanker atau target mekanisme terhadap aktivitas anti kanker hati.Kata kunci: Karsinoma Hepatoseluler, Huh7it-1, Sitotoksisitas, Annona muricataABSTRACTBackground: Hepatocellular carcinoma (HCC) is a malignant tumor of liver cells with prognosis can cause death within 2-3 months. Previous studies of Annona muricata L. on anti-HCV studies at concentrations of 20 μg / mL showed very high toxicity to Huh7it-1 cell line, it was indicated to have anti-cancer potential of liver cells, so this study tested the potency of anticancer activity extract methanol leaf Annona muricata L. (EMDAM) against Hepatocellular Carcinoma Huh7it-1 strain cell line with low dose.Methods: Cells were tested with concentrations of 20, 10, 5, 2.5, 1.25, 0.6, 0.3 μg / mL for 48 hours. The EMDAM cytotoxicity of Huh7it-1 was seen with an inverted microcomputer and then measured with MTT assay [3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium].Results: The results showed that the cells presented non-monolayer form in an inverted microscope with cytotoxicity until the lowest concentration of 0.3 μg / mL reached 84.7%, thus concentrating 50% cytotoxicity (CC50) <0.3 μg / mL.Conclusion: The results indicate that EMDAM has the potential for anti-liver cancer activity. Further studies are needed for purification for active compounds as anticancer or target mechanisms against anti-liver cancer activity.Keywords: Hepatocellular carcinoma, Huh7it-1, Cytotoxicity, Annona muricat

Identification of an antiviral component from the venom of the scorpion Liocheles australasiae using transcriptomic and mass spectrometric analyses

Scorpion venom contains a variety of biologically active peptides. Among them, neurotoxins are major components in the venom, but it also contains peptides that show antimicrobial activity. Previously, we identified three insecticidal peptides from the venom of the Liocheles australasiae scorpion, but activities and structures of other venom components remained unknown. In this study, we performed a transcriptome analysis of the venom gland of the scorpion L. australasiae to gain a comprehensive understanding of its venom components. The result shows that potassium channel toxin-like peptides were the most diverse, whereas only a limited number of sodium channel toxin-like peptides were observed. In addition to these neurotoxin-like peptides, many non-disulfide-bridged peptides were identified, suggesting that these components have some critical roles in the L. australasiae venom. In this study, we also isolated a component with antiviral activity against hepatitis C virus using a bioassay-guided fractionation approach. By integrating mass spectrometric and transcriptomic data, we successfully identified LaPLA₂-1 as an anti-HCV component. LaPLA₂-1 is a phospholipase A₂ having a heterodimeric structure that is N-glycosylated at the N-terminal region. Since the antiviral activity of LaPLA₂-1 was inhibited by a PLA₂ inhibitor, the enzymatic activity of LaPLA₂-1 is likely to be involved in its antiviral activity

Anti-viral activity of Phyllanthus niruri against hepatitis C virus

Hepatitis C virus (HCV) infection is a global problem that causes liver disease and hepatocellular carcinoma. Although the current standard treatment provided a significant improvement on response rate with sustain virology response more than 90%, however, the high cost was remaining limited access to this therapy, resistance emergence and serious side effects which provide the necessities to find the new anti-HCV agents. The current study, we evaluated the ethanol extract of Phyllanthus niruri for its anti-HCV activities. Anti-HCV activity was determined by in vitro culture cells of Huh 7it. Anti-HCV activity of P. niruri extract revealed strong inhibition against HCV with IC50 values of 4.14 µg/mL and yield stronger activity in the entry step of the HCV life cycle. Moreover, the P. niruri extract enhanced anti-HCV activity of simeprevir (NS3 protease inhibitor) with increase the activity up to 4-fold compared to a single treatment of simeprevir. Docking analysis was performed to predict the interaction phyllanthin and hypophyllantin, known compounds of P. niruri against HCV receptor. Both of phyllantin and hypophyllantin were mediated a strong interaction with 4GAG, a protein that involved in entry step of HCV. These results suggested that the ethanol extract of P. niruri may be good candidates for the development of anti-HCV drugs

Another novel subgenotype of hepatitis B virus genotype C from papuans of Highland origin

Hepatitis B virus (HBV) genotypes and subtypes have been identified worldwide. As HBV genotypes/subtypes, the HBV subgenotypes seem to be associated with their geographical distribution and ethnic origin. A previous study showed the novel HBV subgenotype C6 based on the complete genome sequences of isolates in Papua, Indonesia. In the present study, further characterization of HBV in Jayapura (capital of Papua Province), particularly from native people of Papua originating from the highland (highland Papuans) and those from the lowland (lowland Papuans) were examined. Of 32 HBV isolates from both highland and lowland Papuan blood donors with HBsAg positive, part of the S gene and the core gene sequences were analyzed. Analyses of some isolates from highland Papuans were confirmed by the complete genome sequences. Most HBV isolates were classified into genotype C (78.1%), followed by genotype B (18.8%), and genotype D (3.1%). The subtype adr was predominant (71.9%), followed by adw2 (25.1%), and ayw2 (3.1%). As with previous findings, phylogenetic analyses revealed that most HBV isolates from Papuans, C/adr, belonged to subgenotype C6. Interestingly, some C/adr isolates from highland Papuans formed a distinct cluster from all reported subgenotypes of HBV/C, and they differed from HBV/C1‐C10 by 4.2–7.2% over the complete genome. SimPlot analysis showed no evidence of recombination with HBV/C1–C10. The isolated life and closed social systems of highland Papuans, even though some have been moving to Jayapura, likely contribute to the formation of this unique cluster of infection with a novel subgenotype of HBV, named C11

AIRE Functions As an E3 Ubiquitin Ligase

Autoimmune regulator (AIRE) gene mutation is responsible for the development of autoimmune-polyendocrinopathy-candidiasis ectodermal dystrophy, an organ-specific autoimmune disease with monogenic autosomal recessive inheritance. AIRE is predominantly expressed in medullary epithelial cells of the thymus and is considered to play important roles in the establishment of self-tolerance. AIRE contains two plant homeodomain (PHD) domains, and the novel role of PHD as an E3 ubiquitin (Ub) ligase has just emerged. Here we show that the first PHD (PHD1) of AIRE mediates E3 ligase activity. The significance of this finding was underscored by the fact that disease-causing missense mutations in the PHD1 (C311Y and P326Q) abolished its E3 ligase activity. These results add a novel enzymatic function for AIRE and suggest an indispensable role of the Ub proteasome pathway in the establishment of self-tolerance, in which AIRE is involved

AntiHepatitis C Virus Activity of Alectryon serratus Leaves Extract

Hepatitis C Virus (HCV) has infected approximately 2-3% (130-170 million) of the world's population. No vaccine is available to prevent HCV infection. Investigation of anti-HCV agent is thus deemed necessary. Various plants have been explored for their anti-HCV activity. A. serratus is a member of Sapindaceae family, which fruit and seed were traditionally used as insecticide. Anti-HCV activity tested on A.serratus leaves extract has been done. The result showed that leaves extract exhibited anti-HCV with IC50 value of 14.9 μg/ml and 9.8 μg/ml against HCV J6/JFH1 and JFH1a, respectively. The cytotoxicity assay results showed that A.serratus leaves extract was not toxic and has CC50 >100 μg/ml. Mode of action experiment results suggested that A.serratus extract inhibited HCV at the post-entry step. Further fractionation of leaves extract by open column chromatography resulted in 4 fractions. Only Fraction 1 (AP-5F.1) exhibited anti-HCV with IC50 value of 1.2 μg/ml against HCV JFH1a. Separation of AP-5F.1 by open column chromatography resulted in 15 fractions. Fraction number 13 (AP-5F.1.13) exhibited anti-HCV with IC50 value of 0.43 μg/ml against HCV JFH1a. Separation of AP-5F.1.13 by semi preparative-HPLC resulted in isolate identified by TLC and LC-MS method as chlorophyll derivate. There was a possibility that chlorophyll derivate has participated in performing the anti-HCV activity of fractions and extract besides the other compounds contained. In this study, we concluded that A. serratus leaves extract, AP-5F.1, and AP-5F.1.13 exhibited anti-HCV activity against JFH1a virus

ANTIHEPATITIS C VIRUS ACTIVITY OF INDONESIAN MAHOGANY (TOONA SURENI)

 Objective: Toona sureni (Indonesian mahogany) is a member of Meliaceae family and locally known as suren. Previous study reported that T. sureni leaves extract exhibited antiviral activity with 50% inhibitory concentration (IC50) value of 13.9 ± 1.6 μg/ml against hepatitis C virus (HCV) J6/JFH1. Cytotoxicity analysis of T. sureni leaves extract did not reveal any cytotoxicity effect; therefore, further study was taken to investigate the active substances from the extract.Methods: Bioassay-guided isolation of anti-HCV was conducted using Huh-7.5 cells infected with HCV J6/JFH1 in the presence of extracts, fractions, or compounds from the plant.Results: Ethyl acetate fraction (Fr E) exhibited high anti-HCV activity with IC50 value of 1.7 μg/ml. Further, separation of Fr E by open column chromatography resulted in nine sub-fractions (sub-Fr E1-E9). Sub-Fr E3 and E4 have IC50 value of 29.90 μg/ml and 7.68 μg/ml, respectively. Polyphenols compounds have been isolated from sub-Fr E3 and E4. The structures have been determined to be ethyl gallate (1), methyl gallate (2), catechin (3), gallic acid (4), and quercetin 3-O-rhamnoside (5). Among the isolated compounds, gallic acid showed to possess strong anti-HCV activity with IC50 value of 15.9 μg/ml.Conclusion: T. sureni and its isolated compound, gallic acid, may be good candidates to develop for alternative and/or complementary agents of anti-HCV infection