JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2023
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IF 6.1
Xuefeng Wang,Baifei Hu,Haiming Hu,Shuhan Zhou,Mingzhu Yin,Xue Cheng,Zhigang Zhang,Hongtao Liu
Hepatitis B virus (HBV) infection is a serious global health problem that threatens the health of human. Tannic acid (TA), a natural polyphenol in foods, fruits, and plants, exhibits a variety of bioactive functions. In our research, we decide to explore the pharmacological mechanism of TA against HBV replication. Our results showed that TA effectively reduced the content of HBV DNA and viral antigens (HBsAg and HBeAg) in HepG2.2.15 cells. Meanwhile, TA significantly decreased the mRNA expression of HBV RNA, which include total HBV RNA, HBV pregenomic RNA, and HBV precore mRNA. Besides, TA evidently downregulated the activity of HBV promoters in HepG2.2.15 cells. Furthermore, we found that TA upregulated the expression of IL-8, TNF-α, IFN-α, and IFN-α-mediated antiviral effectors in HepG2.2.15 cells. On the contrary, TA downregulated the expression of IL-10 and hepatic nuclear factor 4 (HNF4α). In addition, TA activated the NF-κB and MAPK pathways that contributed to the inhibition of HBV replication. Finally, TA treatment led to the occurrence of autophagy, which accelerated the elimination of HBV components in HepG2.2.15 cells. Taken together, our results elucidated the suppressive effect of TA on HBV replication and provided inspiration for its clinical application in HBV treatment.
研究领域:
病毒学
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药理学
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分子生物学
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细胞生物学
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PubMed
高影响因子
Biochimica et Biophysica Acta-Gene Regulatory Mechanisms
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2021
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IF 4.49
Da Cheng,Cichun Wu,Ying Li,Yao Liu,Juan Mo,Lei Fu,Shifang Peng
Hepatitis B virus (HBV)-associated acute-on-chronic liver failure (ACLF) is a clinical syndrome of severe liver damage. HBV infection is affected by N6-methyladenosine (m 6 A) RNA modification. Here, we investigated whether methyltransferase-like 3 (METTL3)-mediated m 6 A methylation can affect ACLF. Human hepatic cells (THLE-2) were treated with lipopolysaccharide (LPS) to induce cell damage. Proliferation, apoptosis and m 6 A modification were measured by MTT assay , flow cytometry and Dot blot assay. Our results showed that HBV infection significantly enhanced the levels of m 6 A modification and elevated the expression of METTL3 and mature-miR-146a-5p in THLE-2 cells, which was repressed by cycloleucine (m 6 A inhibitor). METTL3 overexpression enhanced m 6 A modification and promoted mature-miR-146a-5p expression. METTL3 overexpression promoted HBV replication and apoptosis, enhanced the levels of pro-inflammatory cytokines, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg), and repressed cell proliferation in THLE-2 cells, which attributed to repress miR-146a-5p maturation. Moreover, a severe liver failure mouse model was established by HBV infection to verify the impact of METTL3 knockdown on liver damage in vivo . HBV-infection led to a severe liver damage and increase of apoptosis in hepatic tissues of mice, which was abolished by METTL3 knockdown. METTL3 knockdown reduced METTL3 expression and impeded miR-146a-5p maturation in HBV-infected mice. In conclusion, this work demonstrates that METTL3 inhibition ameliorates liver damage in mouse with HBV-associated ACLF, which contributes to repress miR-146a-5p maturation. Thus, this article suggests a novel therapeutic avenue to prevent and treat HBV-associated ACLF.
研究领域:
分子生物学
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肝病学
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病毒学
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药理学
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 7
·1-10
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IF 5.833
Wang Lingyun,Yang Junyu,Tang Li,Luo Lianghui,Chen Chunyan,Gong Hang,Cai Changqun
An approach is reported based on the combination of aptamer and metal organic frameworks (MOF) to prepare a molecularly imprinted sensor that recognizes viruses with high specificity and sensitivity. Using MIL-101-NH 2 as a polymer carrier, viral aptamers were introduced into the carrier surface through an amide reaction to specifically identify the target, and surface imprinting is carried out through tetraethyl silicate (TEOS) self-polymerization. The MIL-101-NH 2 is also used as the reference fluorescence signal (λex/λem = 290/460 nm) and rhodamine B as the change signal (λex/λem = 550/570 nm). The ratiometric fluorescence detection and dual recognition strategy not only reduce environmental interference but also greatly improve the sensor’s anti-interference ability, the obtained imprinting factor was 5.72, and the detection limit as low as 1.8 pmol L −1 . Therefore, the molecular imprinting sensor designed realizes the specific and highly sensitive identification of viruses, which provides theoretical support for the application of molecular imprinting technology in clinical diagnosis of viruses.
Aptamer-molecular imprinting polymer based on metal-organic framework ratiometric fluorescent detect virus.
研究领域:
生物化学
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医学诊断
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材料科学
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PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2023
·Vol. 33
·期 7
·2210860
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IF 19
Chenya Zhuo,Huiming Kong,Ke Yi,Chun-Wei Chi,Jiabing Zhang,Ran Chen,Haixia Wang,Caixia Wu,Yeh-Hsing Lao,Yu Tao,Mingqiang Li
Chronic hepatitis B infection remains incurable due to the stable presence of various forms of hepatitis B virus (HBV) genome, especially the HBV covalently closed circular DNA (cccDNA). The emergence of clustered regularly interspaced short palindromic repeat (CRISPR) technology provides a new opportunity to potentially cure the HBV infection. However, the efficiency and specificity remain unsatisfactory, especially for nonviral CRISPR/Cas9 delivery. To tackle these, a liver-specific CRISPR/Cas9 magnetic nanosystem FMNP pAG333/sgXPP is constructed based on fluorinated polyethylenimine-coated magnetic nanoparticles and liver-specific ApoE.HCR.hAAT promoter-driven Cas9-T2A-EGFP plasmid with dual sgRNAs. The elaborate system enables magnetic field-induced spatially specific distribution and hepatocyte-specific promoter-driven liver-specific gene editing. Moreover, this CRISPR/Cas9 magnetic nanosystem is designed to disrupt the two conserved sites in X opening reading frame and Pol opening reading frame of the HBV genome, thereby significantly inactivating the HBV genome without showing off-target effects. Treatment with FMNP pAG333/sgXPP for 7 days reduces serum HBsAg levels by 76% with a total editing efficiency of ≈20% in the two conserved sites. Collectively, this study demonstrates spatiotemporal liver genome editing as well as the feasibility of applying a nonviral CRISPR/Cas9 vector for HBV treatment, which may open up a new application for CRISPR therapeutics.
研究领域:
分子生物学
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纳米技术
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病毒学
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治疗学
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高影响因子
ANALYST
·
2021
·Vol. 146
·期 12
·3924-3932
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IF 4.616
Siyu Chen,Lianghui Luo,Lingyun Wang,Chunyan Chen,Hang Gong,Changqun Cai
Highly selective and highly efficient identification of large viruses has been a major obstacle in the field of virus detection. In this work, a novel sandwich resonance light scattering sensor was designed based on molecularly imprinted polymers (MIPs) and aptamers for the first time. One of the recognition probes was obtained by molecular imprinting using environmentally friendly carbon spheres as carriers and the other by modification of the aptamer that can specifically recognize hepatitis B virus (HBV) on the surface of silicon spheres. In the presence of both probes, an MIP-HBV-aptamer sandwich structure was formed continuously in the system with the increase in HBV concentration, resulting in a strong resonance light scattering response. Finally, satisfactory selectivity and sensitivity were obtained, and the imprinting factor was as high as 7.56, which was higher than that reported in previous works of viral molecular imprinting sensor. In addition, it is of great significance to solve the problem of insufficient selectivity of traditional detection methods for macromolecular targets.
研究领域:
分析化学
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生物技术
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病毒学
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PubMed
高影响因子
TALANTA
·
2026
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IF 6.7
Mingqi Yang,Jiali Hu,Zhenlin Zhang,Liting Huang,Yahan Yu,Dexing Qiu,Yexin Zu,Yanyan Liu,Zhenyu Lin
Infection with hepatitis B virus (HBV) represents a major challenge to global health, especially in areas of inadequate healthcare infrastructure, rapid and on-site detection plays a crucial role in effective disease management and control. Consequently, it is necessary to develop some simple but sensitive HBV screening techniques. In this work, a homogeneous electrochemical biosensor was constructed for sensitive and specific HBV DNA detection through the integration of CRISPR/Cas12a target recognition and magnetic bead-based separation. The sensor exploits the collateral cleavage activity of CRISPR/Cas12a protein upon recognition of target double-stranded DNA, enabling the degradation of methylene blue-labeled single-stranded DNA captured on magnetic beads (MBs). Then the MBs are separated and redissolved in the solution and the electrochemical response of the system can be tested in homogeneous solution. The electrochemical signal has relationship with the concentration of HBV DNA within a range of 10 fM - 10 nM, and the LOD is 3.74 fM. The sensor demonstrates excellent selectivity and biological stability, which has potential application in clinical diagnosis, especially in resource-limited environments.
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高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2026
·
IF 2.5
Han Liu,Zhuo Li,Xilin Zhu,Ying Liu,XiaoPan Wu
Background & aims Caspase-10 (CASP10) participates in death receptor–mediated apoptosis, but the relevance of the CASP10 coding variant rs13006529 (T > A; p.Leu522Ile) to PEG-IFN-α response in chronic hepatitis B (CHB) is unknown. We assessed this association and whether differential apoptotic activity explains variant-associated HBV burden. Methods rs13006529 was genotyped in 262 patients receiving PEG-IFN-α with or without nucleos(t)ide analogues. Allele and genotype frequencies were compared by χ 2 tests; sustained virological response (SVR) was analyzed by univariable and multivariable logistic regression. HBV-infected HepG2-NTCP cells expressing empty vector or either CASP10 variant were compared; variant-expressing cells received vehicle or zVAD-fmk. Apoptosis was assessed by flow cytometry, caspase-3/7 activity, and Western blotting. HBV total RNA and pgRNA, intracellular DNA, cccDNA, and HBsAg/HBeAg were measured by RT-qPCR, qPCR, Southern blotting, and ELISA, respectively. Results The A allele was more frequent in patients without than with SVR (30.6% vs 17.1%; P = 0.002) and independently associated with lower odds of SVR (adjusted OR per A allele, 0.50; 95% CI, 0.27–0.86; P = 0.017). SVR rates were 36.9% in TT and 19.8% in AT/AA carriers (P = 0.003). Leu522-expressing cells showed greater apoptosis and lower HBV total RNA, pgRNA, intracellular DNA, and cccDNA than Ile522-expressing cells. zVAD-fmk attenuated the between-variant difference in apoptosis and partially narrowed the corresponding differences in HBV RNA and intracellular DNA. Conclusions rs13006529 was independently associated with PEG-IFN-α response. Differential caspase-dependent apoptotic activity partly accounts for variant-associated HBV burden differences.
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高影响因子
ANTIVIRAL RESEARCH
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2026
·
IF 4.3
Pu Wang,Xinchang Liu,Yong Xie,Yingjun Zhang,Baohua Gu,Jing Li,Yunfu Chen
CAM-A (GLS4) rapidly suppresses HBV DNA with delayed HBsAg decline and ALT flares. • CAM-A upregulates ISGs and activates IFN-α/γ pathways in HBV models. • CAM-A induces TUNEL + caspase-3-independent death of HBcAg + hepatocytes. • CAM-A effects persist without adaptive immunity; synergize with innate immunemodulators such as TLR7 agonist. Capsid assembly modulators (CAMs) are promising antiviral agents for chronic hepatitis B virus (HBV) infection. This study investigates the anti-HBV activity and immunomodulatory effects of Class I CAMs (CAM-A) in vitro and in vivo . In HepAD38 cells, CAM-A treatment significantly upregulated the expression of interferon-stimulated genes (IP-10, MX1, OAS1). In AAV-HBV mice, CAM-A (GLS4) induced rapid reductions in HBV DNA and delayed HBsAg decline accompanied by transient ALT elevation, upregulated interferons and Tnfsf10/Tnfrsf10b gene expression, while terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and immunohistochemistry (IHC) experiments indicated caspase-3-independent cell death in HBV-infected hepatocytes. Importantly, the antiviral and HBsAg-lowering effects of CAM-A did not require adaptive immunity. Furthermore, combination of CAM-A with a TLR7 agonist in AAV-HBV mice, or with Peg-IFN-α in primary human hepatocytes (PHH) resulted in synergistic antiviral effects, underscoring the therapeutic potential of these agents to promote sustained HBsAg decline and support functional cure strategies in chronic HBV infection.
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高影响因子
CHEMICO-BIOLOGICAL INTERACTIONS
·
2024
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IF 4.7
Hong Ding,Xuanxiao Lu,Xiaoguo Ji,Shijie Wang,Jiayang Jin,Mengyao Zhao,Xiaofeng Hang,Liming Zhao
Selenium supplements are beneficial to human health, however, concerns regarding the toxicity of inorganic selenium have stimulated research on safer organic compounds. The main objective of this study was to develop a novel glucosamine-selenium compound (Se-GlcN), clarify its structure, and subsequently investigate its oral toxicity and in vitro anti-hepatitis B virus (HBV) activity. Electron microscopy, infrared, ultraviolet spectroscopy, nuclear magnetic resonance and thermogravimetric analyses revealed a unique binding mode of Se-GlcN, with the introduction of the Se-O bond at the C6 position, resulting in the formation of two carboxyl groups. In acute toxicity studies, the median lethal dose (LD 50 ) of Se-GlcN in ICR mice was 92.31 mg/kg body weight (BW), with a 95% confidence interval of 81.88 to 104.07 mg/kg BW. A 30-day subchronic toxicity study showed that 46.16 mg/kg BW Se-GlcN caused livers and kidneys damage in mice, whereas doses of 9.23 mg/kg BW and lower were safe for the livers and kidneys. In vitro studies, Se-GlcN at 1.25 μg/mL exhibited good anti-HBV activity, significantly reducing HBsAg, HBeAg, 3.5 kb HBV RNA and total HBV RNA by 45%, 54%, 84%, 87%, respectively. In conclusion, the Se-GlcN synthesized in this study provides potential possibilities and theoretical references for its use as an organic selenium supplement.
研究领域:
药物化学
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毒理学
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病毒学
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药理学
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DOI
PubMed
高影响因子
RSC Advances
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2015
·Vol. 5
·期 55
·44714-44721
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IF 3.84
Siqi Zhang,Kun Wang,Zhenyu Li,Zhongmin Feng,Ting Sun
In this work, a one-pot signal amplified strategy was constructed based on generating a hemin/G-quadruplex horseradish peroxidase-mimicking DNAzyme as the color product for the detection of DNA sequences. A DNA sequence associated with the hepatitis B virus (HBV) was selected as a model target. The enzyme-free and label-free biosensor contained three oligonucleotide terms as target DNA, hairpin structures H1 and H2 which were partially complementary. It should be noted that the G-quadruplex structure was partially hidden in the hairpin structure H2. In the absence of THBV, hairpins H1 and H2 were stable enough due to the complementary sequences at the end of the oligodeoxynucleotides, which made it difficult to complete the self-assembly. Upon addition of THBV, self-assembly of THBV with H1 allows the rest of the DNA sequence of H1 to facilitate H1–H2 complex formation and releases THBV to the next cycle. In the presence of hemin and K+, the G-quadruplex at the end of the H1–H2 complex was liberated to form a hemin/G-quadruplex structure, which could catalyze achromatous tetramethylbenzidine (TMB) into a colored product. The color change of the solution could be quantitated by spectrophotometry and the naked eye. We also employed 20% ethanol in the buffer to accelerate H1–H2 complex formation in the DNA strand displacement reaction (DSDR), which cut down the reaction time of the detection process from 12 h to 1 h. The detection limit of the colorimetric sensor is 9.5 pM, and the linear range is 50 pM to 100 nM.
研究领域:
生物化学
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分子生物学
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分析化学
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医学诊断
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DOI
高影响因子
ARCHIVES OF VIROLOGY
·
2024
·Vol. 169
·期 11
·1-10
·
IF 2.5
Jiao Qian,Zhu Shu,Liao Baolin,Liu Huiyuan,Guo Xiaoyan,Wu Lina,Chen Chuming,Peng Liang,Xie Chan
NLR family member X1 (NLRX1) is an important member of the NOD-like receptor (NLR) family and plays unique roles in immune system regulation. Patients with hepatitis B virus (HBV) infection are more likely to have the NLRX1 mutation p.Arg707Cys than healthy individuals. It has been reported that NLRX1 increases the infection rate of HBV in HepG2 cells expressing sodium taurocholate cotransporting polypeptide (NTCP). However, the role of NLRX1 mutation ( p.Arg707Cys ) in hepatitis remains unclear. We constructed Huh7 cells that stably overexpressed NTCP, using LV003 lentivirus. First, wild-type (WT) and mutant (MT) NLRX1 overexpression plasmids were constructed. The MT plasmid contained a point mutation at position 707 of the WT overexpression plasmid. Then, Huh7-NTCP cells were transfected with the WT or MT NLRX1 overexpression plasmid, and subsequent NLRX1 expression was analyzed using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. HBV RNA levels were determined using RT-qPCR. HBsAg and HBcAg levels were confirmed immunohistochemically. Interferon alpha (IFN-α), interleukin 6 (IL-6), and type I interferon beta (IFN-β) levels were determined using enzyme-linked immunosorbent assay kits. p-p65, p-interferon regulatory factor (IRF) 3, and p-IRF7 expression levels were examined using western blot. The interaction of NLRX1 and retinoic acid-inducible gene (RIG)-1/mitochondrial antiviral signaling (MAVS) protein was confirmed by coimmunoprecipitation. The interaction of NLRX1 with IFN-α, IL-6, or IFN-β was analyzed by dual luciferase reporter gene assay. The levels of HBV RNA, HBsAg, and HBcAg in infected cells transfected with the WT NLRX1 or MT NLRX1 expression plasmid were higher than those in the untransfected control group; and these levels were lower in the cells transfected with MT NLRX1 than in those transfected with WT NLRX1. The levels of IFN-α, IFN-β, IL-6, p-p65, p-IRF3, and p-IRF7 were lower in cells transfected with WT NLRX1 or MT NLRX1 than in control cells. The levels of IFN-β, p-p65, p-IRF3, and p-IRF7 were higher in cells transfected with MT NLRX1 than in those transfected with WT NLRX1. Moreover, NLRX1 competitively inhibited RIG1 binding to MAVS, but the mutation in MT NLRX1 reduced this inhibitory effect. In addition, NLRX1 decreased the promoter activity of IFN-α, IFN-β, and IL-6. Our findings revealed that NLRX1 is a regulatory factor that inhibits the anti-HBV ability of hepatocytes and that the mutation p.Arg707Cys in NLRX1 suppresses HBV infection and activates the IFN/nuclear factor κB pathway.
研究领域:
免疫学
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病毒学
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分子生物学
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细胞生物学
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PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2022
·
IF 5.195
Shiqi Zhu,Haimei Wen,Wenling Wang,Yong Chen,Fengmei Han,Wentao Cai
Ethnopharmacological relevance Salvia miltiorrhiza (the roots of S. miltiorrhiza Bunge, Danshen in Chinese), a traditional Chinese medicine , has been clinically used to prevent and treat various diseases, such as cardiovascular and cerebrovascular diseases , diabetes, and hepatitis B, in China and some other Asian countries. Lithospermic acid (LA), a polyphenol derived from S . miltiorrhiza , has been reported to exhibit multiple pharmacological properties, such as anti-inflammatory, anti-HIV, and anti-carbon tetrachloride-induced liver injury activities. However, little is known about the anti-hepatitis B virus (HBV) activity of LA. Aim of the study The study was projected to investigate the anti-HBV activity of LA in vitro (HepG2.2.15 and pHBV1.3-transfected HepG2 cells) and in vivo (pAAV-HBV1.2 hydrodynamic injection [HBV-HDI] mice) and explore the potential mechanism as well. Materials and methods Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) contents were detected by ELISA kits. HBV DNA and hepatitis B core antigen (HBcAg) levels were evaluated by quantitative real-time polymerase chain reaction and immunohistochemistry assay, respectively. The proteins in autophagy process, lysosomal acidic function, and autophagy-related signaling pathways were examined by Western blot . Transmission electron microscopy was used to observe the number of autophagosomes and autolysosomes. Confocal microscopy was applied to analyze the autophagic flux and lysosomal acidification, using mCherry-enhanced green fluorescent protein (EGFP)-microtubule-associated protein light chain (LC)3 and lysosomal probes, respectively. Results LA exhibited anti-HBV activity by inhibiting HBV DNA replication in HepG2.2.15 and pHBV-transfected HepG2 cells in dose- and time-dependent manners and hampering HBsAg and HBeAg levels in HepG2.2.15 cells to a certain extent. LA reduced HBV DNA, HBsAg/HBeAg, and HBcAg levels in the serum/liver tissues of HBV-HDI C57BL/6 mice during the 3-week treatment and suppressed the withdrawal rebound of HBV DNA and HBsAg in the mice serum. LA increased LC3-II protein expression and the number of autolysosomes/autophagosomes and promoted the degradation of sequestosome 1(p62) protein in vitro and in vivo . LA enhanced the co-localization of LC3 protein with autolysosomes, further confirming the ability of LA to induce a complete autophagy. Knockdown of autophagy-related gene (Atg) 7 or 5 in vitro and administration of 3-methyladenine (an autophagic inhibitor) in vivo disabled the inhibitory efficacy of LA on HBV DNA replication, suggesting that the anti-HBV efficacy of LA depended on its ability of inducing autophagy. LA could enhance lysosomal acidification and improve the function of lysosomes by promoting the protein expression of lysosomal-associated membrane protein (LAMP)-1, LAMP-2, and mature cathepsin D , which may contribute to the autophagic induction of LA. LA inhibited the activation of AKT and mammalian target of rapamycin (mTOR) induced by HBV, which was reversed by IGF-1 (an agonist of the PI3K/AKT/mTOR signaling pathway), indicating that LA elicited autophagy through hampering the PI3K/AKT/mTOR signaling pathway. Conclusion We revealed the anti-HBV activity and mechanism of LA in vitro and in vivo . This study facilitates a new understanding of the anti-HBV potent components of S. miltiorrhiza and sheds light on LA for further development as an active constituent or candidate used in the therapy against HBV infection.
研究领域:
药理学
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病毒学
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分子生物学
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中医药
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PubMed
高影响因子
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