共查询到20条相似文献，搜索用时 31 毫秒
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AIM：To investigate the antiviral effect of hepatocyte-targeting and cell-penetrating peptide and Musca domestica cecropin (HTPP-MDC) fusion polypeptide against hepatitis B virus (HBV) in vitro, and to observe the penetrating ability of HTPP-MDC in hepatocytes. METHODS： HepG2.2.15 cells and Chang liver cells were co-cultured in vitro with HTPP-MDC at different concentrations. MTT assay was used to detect the cytotoxicity of HTPP-MDC in vitro. The levels of HBsAg, HBeAg and HBV DNA in HepG2.2.15 cell culture supernatants were measured by ELISA and real-time fluorescence quantitative PCR. The penetrating ability of HTPP-MDC was detected by laser confocal microscopy. RESULTS：The levels of HBsAg, HBeAg and HBV DNA in the supernatants of HepG2.2.15 cells treated with HTPP-MDC were remarkably reduced. The inhibitory effect of HTPP-MDC depended on the dose and action time of the drug. FITC-labeled HTPP-MDC was observed inside the cells under laser confocal microscope. CONCLUSION：HTPP-MDC strongly inhibits HBV replication in HepG2.2.15 cells. The penetrating ability of HTPP-MDC into hepatocytes indicates that HTPP-MDC is useful in clinic therapy for chronic hepatitis B-related diseases in the future. 相似文献
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AIM:To investigate the effects of xeroderma pigmentosum D (XPD) and p53 on the replication of hepatitis B virus (HBV). METHODS:Recombinant plasmid pEGFP-N2/XPD and vacant vector plasmid pEGFP-N2 were transfected into HepG2.2.15 cells by liposome. On the next day, these cells were incubated with pifithrin-α, a p53 inhibitor, at a concentration of 20 μmol/L for 24 h. The cells were divided into 5 groups: blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, pEGFP-N2/XPD+pifithrin-α group and pifithrin-α group. The mRNA expression of XPD, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus X protein (HBx) was detected by RT-PCR. The content of HBsAg and HBeAg in the supernatants of culture medium was measured by ELISA. The content of HBV-DNA in the supernatants of culture medium was examined by fluorescence quantitative PCR. Using the method of bDNA, the content of HBV-DNA in the core particles was assessed. RESULTS:The expression of XPD mRNA was elevated by the transfection of recombinant plasmid pEGFP-N2/XPD. The increase in XPD expression significantly down-regulated the mRNA expression of HBsAg, HBeAg and HBx. The content of HBsAg and HBeAg in the supernatants of culture medium was significantly decreased by the increase in XPD expression. The results of fluorescence quantitative PCR showed that the content of HBV-DNA in the supernatants of culture medium was significantly down-regulated by the increase in XPD expression. bDNA results showed that the content of HBV-DNA in the core particles was significantly decreased by the increase in XPD expression.Pifithrin-α abolished the above-mentioned effects of XPD (all P<0.01). CONCLUSION: XPD inhibits the replication of HBV through p53 pathway. Therefore, XPD and p53 may be the targets for antiviral therapy of hepatitis B. 相似文献
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KONG Xiang-li ZHANG Ping HE Fang ZHANG Min TANG Hong FENG Yun WU Qi HUANG Ning WANG Bo-yao 《园艺学报》2009,25(8):1606-1611
AIM: We previously demonstrated that HMGN2 is an antibacterial effector molecule in human LAK cells. This study was to examine the antiviral activity of HMGN2 against human hepatitis virus.METHODS: The purification and identify of HMGN2 proteins including preparative acid-urea polyacymide gel electrophoresis elution, reverse-phase high-performance liquid chromatography, mass spectrum, Western blotting and antimicrobial assay were conducted. The cellular toxicity of HMGN2 to the HepG2.2.15 cells was detected by MTT assay. HBeAg and HBsAg expressions were measured by ELISA. HBV DNA copies were determined by real time quantitative PCR.RESULTS: A bulk of HMGN2 was isolated and purified from the acid soluble proteins of human lymph node, and identified. The HBV-transfected HepG2.2.15 cell line was used in the in vitro assay system.In the range of testing 1-100 mg/L of HMGN2, no cytotoxicity to HepG2.2.15 cells was detected by MTT assay.When incubated with HMGN2 at 1-5 mg/L for 72 h or 144 h, a significant reduction in HBeAg and HBsAg expression and in HBV DNA copies was observed in the supernatant of HepG2.2.15 cells. CONCLUSION: HMGN2 protein markedly inhibits HBV expression and replication in vitro. 相似文献
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Effect of proprotein convertases on transforming growth factor β1-induced HBV replication inhibition
AIM: To study the effect of proprotein convertases (PCs) on the transforming growth factor (TGF) β1-induced inhibition of HBV replication.METHODS: HepG2.2.15 cells cultured regularly were exposed to recombinant TGFβ1 at concentration of 2 μg/L or 5 μg/L and/or PC inhibitor at concentration of 20 μmol/L for 18 h. The total RNA and HBV core particle DNA were extracted from these cells, and PC mRNA and core-associated HBV DNA were detected by real-time PCR technique. RESULTS: The mRNA expression levels of 7 PCs in HepG2.2.15 cells were observed with various degrees. Recombinant TGFβ1 significantly up-regulated the mRNA expression of all PCs except for the down-regulation of PC5/6, though PC1/3 and PC2 were up-regulated most obviously. Furin and PACE4 were the predominant PCs before and after TGFβ1 exposure when the basic mRNA expression was taken into account. Further study showed that TGFβ1-induced the inhibition of HBV replication was abrogated by PC inhibitor in HepG2.2.15 cells. CONCLUSION: TGFβ1-induced the inhibition of HBV replication is mediated by the up-regulation of PCs, which might be of many implications in efficient interferences of TGFβ1 on HBV replication. 相似文献
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SHI Cheng-gang HU Ai-ling ZENG Xiao-lin YIN Qiong-li LI Mei CHENG Gui-feng 《园艺学报》2013,29(10):1815-1820
AIM：To detect the expression of preS1/S2 antigen (preS1/S2-Ag) and other antigens of hepatitis B virus (HBV) in renal tissues of patients with HBV-associated glomerulonephritis (HBV-GN), and to analyze their roles in the diagnosis of HBV-GN.
METHODS：Patients hospitalized in our department from January in 2003 to January in 2013 were retrospectively studied. A total of 49 patients with positive HBV surface antigen (HBsAg) serology, clinical manifestations of hematuria and/or proteinuria, and pathological diagnosis of glomerulonephritis, without systemic lupus erythematosus, anaphylactic purpura, diabetes or hepatitis C, were selected. PreS1/S2-Ag, HBV e antigen (HBeAg), HBsAg and HBV core antigen (HBcAg) in the renal tissues were examined. Five cases of glomerular minimal-change disease (MCD) with negative HBsAg and 5 cases of non-glomerulonephritis with positive HBsAg served as controls. RESULTS：The positive rates of preS1/S2-Ag, HBeAg, HBsAg and HBcAg in the renal tissues from the 49 patients of glomerulonephritis with HBV infection were 32.7% (16 cases), 38.8% (19 cases), 14.3% (7 cases) and 46.9% (23 cases), respectively. Total antigen positive rate was 70.2% (36 cases). The expression of preS1/S2-Ag was located in the cytoplasm of renal tubular epithelial cells, glomerular epithelial cells, endothelial cells and mesangial cells, and positively correlated with the expression of HBcAg (r=0.459, P<001). The 4 antigens were not detected in the 5 cases of HBsAg-negative patients with glomerular MCD. In the 5 cases of HBsAg-positive patients with non-glomerulonephritis, there were 2 cases expressing HBeAg and 1 case expressing HBcAg, but no cases expressing preS1/S2-Ag or HBsAg. CONCLUSION：The expression of preS1/S2-Ag in renal tissues suggests that HBV may invade the cells of renal tissue. Combined detection of the 4 antigens could elevate the rate of diagnosis of HBV-GN. 相似文献
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AIM: To study the effect of hepatitis virus B proteins on peripheral blood mononuclear cells (PBMCs) from patients among various types of chronic hepatitis B virus (HBV) infection.METHODS: 80 patients of various types of chronic HBV infection were observed, including 40 HBeAg positive with abnormal alanine aminotransferase (ALT) (A group), 20 HBeAg positive with persistent normal ALT(B group), 20 HBeAg and HBV-DNA negative with persistent normal ALT level(C group). IL-10, IFN-γ in CD8+CD28+T cells, after stimulation with PHA, HBeAg and HBcAg for 48 h, were inspected respectively in PBMCs.RESULTS: IFN-γ was significantly lower in HBeAg positive patients. IL-10 was significantly higher in HBeAg positive with normal ALT. CD8+CD28+T were significantly lower than others. CONCLUSION: In HBeAg positive group, secretion of cytotoxic T lymphocyte (CTL) and Th1 type cellular immunologic reaction is decreased, Th2 type cellular immunologic reaction is enhanced. 相似文献
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LI Jin-ge ZHOU Yong-xing LIAN Jian-qi JIA Zhan-sheng FENG Zhi-hua BAI Xian-guang 《园艺学报》2000,16(9):792-796
AIM: To observe the inhibition of HBc/HBeAg expression in the 2.2.15 cell transfected by two-unit ribozyme. METHODS: By use of subclone technique, two-unit ribozyme gene which was cutted from pGEM-Rz123 was ligated into the eukaryotic expression vector pBBS212. The recombinant plasmid was cotransfected into 2.2.15 cell using lipofectamine. Ribozyme was detected by dot-blot hybridization. The s and e/c antigen of HBV were detected by using ELISA, immunohistochemical technique, image analysis system and Western blot. RESULTS: After the transfected cell was selected two weeks by hygromycin B and G418. We found by dot-blot hybridization that ribozyme can express on 2.2.15 cell. HBeAg level can be reduced by 48.6% in the transfected 2.2.15 cell with two-ribozyme. Using immunohistochemical technique and image analysis system, Western blot, we observed that the level of HBcAg expressed in endocellular went down. CONCLUSION:Al these results strongly indicate that two-unit ribozyme can inhibit hepatitis B virus expression in the cell. 相似文献
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ZHOU Hong-zhong LIU Bo REN Ji-hua TAO Na-na CHEN Xiang LI Wan-yu CHEN Juan 《园艺学报》2016,32(8):1425-1429
AIM: To investigate the role of heat shock protein 70(HSP70)in hepatitis B virus (HBV) replication. METHODS: The effect of HBV replication on the expression of HSP70 was analyzed by RT-qPCR. The overexpression efficiency of HSP70 was confirmed by Western blot. The effect of HSP70 overexpression on HBV DNA replicative intermediates was analyzed by RT-qPCR and Southern blot. The effects of HSP70 overexpression on the expression level of HBV 3.5 kb mRNA and HBV core protein were measured by RT-qPCR and Western blot, respectively. The Effect of HSP70 overexpression on HBV promoter activity was detected by dual luciferase reporter system. RESULTS: The mRNA levels of HSP70 were inhibited by HBV replication. Overexpression of HSP70 repressed the expression of HBV DNA replicative intermediates, 3.5 kb mRNA and core protein, as well as HBV core promoter activity. CONCLUSION: HBV replication inhibits the expression of HSP70. Overexpression of HSP70 represses HBV replication. These data suggest that HSP70 repressed HBV replication by inhibiting HBV core promoter activity. 相似文献
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AIM: To assess effects of atorvastatin (Ator) on cardiac myocytes (CM) hypertrophy of neonatal rat induced by angiotensinⅡ (AngⅡ) in vitro and Toll like receptor 4 (TLR4) expression for theoretical bases of preventing and treating myocardial hypertrophy.METHODS: CM of neonatal Sprague-Dawley (SD) rats were isolated with trypsin digestion method and those growth-arrested cells were stimulated with 10-7 mol/L AngⅡ in the presence of various concentrations of Ator.The method of coomassie brilliant blue was adopted to evaluate the protein contents of CM.The changes in β-MHC,AT1 receptor and TLR4 mRNA expression were observed by RT-PCR.RESULTS: ① AngⅡ increased the protein content of CM and β-MHC mRNA expression significantly,upregulated AT1 receptor and TLR4 gene expression.② In a dose-dependent manner,Ator inhibited the increases in the protein contents of CM and β-MHC mRNA expression induced by AngⅡ.③ In a dose-dependent manner,Ator downregulated the AT1 receptor and TLR4 mRNA expression of CM hypertrophy of neonatal rat induced by AngⅡ.CONCLUSION: Ator inhibits CM hypertrophy,downregulates the AT1 receptor and TLR4 gene expression. 相似文献
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YANG Ping L&# Shang-rui LI Wen-ling ZHANG Yu-qin PAN Qi XU Meng-yao YANG Bo HU Ya-e 《园艺学报》2019,35(7):1153-1162
AIM:To evaluate the expression and biological role of Toll-like receptor 4 (TLR4) in human non-small-cell lung cancer (NSCLC) cells. METHODS:The mRNA and protein levels of TLR4 in NSCLC tissue were exa-mined by RT-qPCR, Western blot, and immunohistochemistry. After treating the A549 cells and SPC-A-1 cells with TLR4 stimulator lipopolysaccharide (LPS) and inhibitor TAK-242, RT-qPCR, Western blot and flow cytometry were performed to detect the expression of TLR4. The migration and invasion abilities were detected by Transwell assay, and the mRNA expression of matrix metalloproteinase (MMP)-2, MMP-9, and vascular endothelial growth factor (VEGF) was also detected. RESULTS:The mRNA and protein levels of TLR4 were higher in the NSCLC tissue than those in the noncancerous tissue (P<0.01). LPS stimulation significantly increased the mRNA and protein expression levels of TLR4 in the NSCLC cell lines A549 and SPC-A-1 (P<0.01). The LPS-induced TLR4 activation enhanced the migration and invasion abilities of A549 cells and SPC-A-1 cells (P<0.01). LPS increased the expression levels of MMP-2, MMP-9 and VEGF in the A549 cells and SPC-A-1 cells (P<0.01). Moreover, the expression levels of TLR4, MMP-2, MMP-9 and VEGF, as well as the migration and invasion abilities of the cells were blocked by TAK-242 (P<0.01). CONCLUSION:TLR4 might be involved in the migration and invasion of NSCLC cells, and TLR4 inhibition might be considered as a therapeutic target for treatment of NSCLC. 相似文献
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AIM: To study the effect of NF-κB decoy oligodeoxynucleotides(ODNs) on TLR4 and IL-8 expression in LPS-induced SW480 cells. METHODS: SW480 cells were cultured in vitro and stimulated for 3 h with LPS (10 μg/L). NF-κB decoy oligodeoxynucleotides mediated by lipofectin 2000 were added into the cell culture for 6 h. The supernatants were collected and messured for IL-8 by ELISA. TLR4 mRNA and IL-8 mRNA were examined by RT-PCR, respectively. The results were compared with control group, Scrambled ODNs group and lipofectin 2000 group. RESULTS: After SW480 cells were stimulated by LPS, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly increased, and the difference compared with control group was obvious. After treated with NF-κB decoy oligodeoxynucleotides, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly inhibited. The Scrambled ODNs group and lipofectin 2000 group had no effect on them. CONCLUSION: NF-κB decoy ODNs will become a new gene drug for treating inflammatory bowel disease(IBD). 相似文献
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AIM：To investigate the effects of shRNA-mediated seven in absentia homolog 2 (SIAH2; one of ubiquitin ligases) gene silencing on cell cycle and apoptosis of human hepatoma cell line HepG2. METHODS：The specific recombinant vector pGenesil-SIAH2 was transiently transfected into HepG2 cells with Lipofectamine 2000. RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of SIAH2. MTS assay was employed to evaluate the cell proliferation. Flow cytometry was used to determine the cell cycle and apoptosis of the transfected cells. RESULTS：Compared with control groups, the mRNA and protein levels of SIAH2 were reduced by pGenesil-SIAH2 transfection in HepG2-SIAH2 group. The proliferation of HepG2-SIAH2 cells was significantly inhibited. The percentage of G1-phase cells and the early apoptotic rate were significantly higher in HepG2-SIAH2 cells. CONCLUSION: Tansfection of pGenesil-SIAH2 effectively inhibits the proliferation of HepG2 cells, arrests the cells in G1 phase and induces apoptosis, indicating an experimental basis of SIAH2-targeting gene therapy for hepatocellular carcinoma. 相似文献
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LI Wei CAO Ji ZHOU Ling-li LUO Wang YANG Chun LUO Cheng-piao LI Yuan SU Jian-jia 《园艺学报》2014,30(12):2142-2147
AIM: To investigate the effect of silencing cell division cycle 25a (CDC25a) gene on the proliferation of human hepatoma HepG2 cells. METHODS: CDC25agene in human hepatoma HepG2 cells was silenced by RNA interference. Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells. Western blotting was applied to detect the expression of CDC25a at protein level. In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells. RESULTS: The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P＜0.05). The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P＜0.05). The cell proliferation in silence group was lower than that in negative control group and normal control group (P＜0.05). The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase. CONCLUSION: Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effectively inhibits the CDC25agene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25agene may be a key target for the treatment of liver cancer. 相似文献
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HAI Guang-fan NIU Bing-xuan LI Pin-pin GUO Lan-qing CUI Tai-zhen LI Sheng-ying 《园艺学报》2014,30(10):1879-1882
AIM:To investigate the effects of nodosin extracted from Chinese traditional medicine on the proliferation of HepG2 cells cultured in vitroand to detect the protein expression of Bcl-2 and Bax in HepG2 cells. METHODS:HepG2 cells were treated with different concentrations (1.25, 2.5, 5, 10 and 20 μmol/L) of nodosin for 24 h. The morphological changes of HepG2 cells were observed under inverted microscope. The inhibitory rates of HepG2 cell growth were detected by MTT assay. The apoptotic rates and the protein expression of Bcl-2 and Bax were analyzed by flow cytometry. RESULTS:Shrunken and suspended HepG2 cells increased with the increases in the concentrations of nodosin. The apoptotic rates and the expression of Bax increased with the increases in the doses of nodosin, while the expression of Bcl-2 decreased. CONCLUSION: Nodosin inhibits the growth of HepG2 cells in a dose-dependent manner. The inhibition of HepG2 cell growth is induced by decreasing Bcl-2 and increasing Bax, thus promoting cell apoptosis. 相似文献
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今年是《园艺学报》创刊发行50周年。50年来,《园艺学报》坚持为学术交流服务,为促进学科发展作贡献的办刊原则,以"科学性;创新性;对生产和科研有参考启迪作用"的标准,收录和发表了大量高水平的论文,记载了几代科技工作者呕心沥血创新之作,反映了中国园艺科学技术和园艺产业的发展历程。 相似文献
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AIM: To explore whether miR-21 low expression enhances the effect of matrine (MAT) on the apoptosis of hepatocellular carcinoma cells.METHODS: Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of miR-21 in the HepG2 cells treated with different concentrations of MAT. The effect of miR-21 on MAT-induced HepG2 cell apoptosis was analyzed by flow cytometry. The mRNA and protein expression of Bcl-2 and Bax in the HepG2 cells treated with MAT was determined by RT-qPCR and Western blot.RESULTS: The expression of miR-21 increased with the increasing concentration of MAT. Low expression of miR-21 promoted MAT-induced apoptosis, and enhanced the expression of Bax at mRNA and protein levels (P<0.05), while inhibited the expression of Bcl-2 at mRNA and protein levels (P<0.05).CONCLUSION: Low expression of miR-21 enhances MAT-induced HepG2 cell apoptosis by inhibiting the expression of Bcl-2 and promoting Bax expression. 相似文献