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asialoglycoprotein receptor and liposome synergistically mediate the gene transfer into primary rat hepatocytes

Science China(Life Sciences) 1999年第4期42卷 435-440页

作者：李崇辉温守明翟海峰孙曼霁 1. Institute of Pharmacology and Toxicology Academy of Military Medical Sciences 100850 Beijing China 2. Department of Clinic Pharmacology General Hospital of Air Force 100036 Beijing China 3. Institute of Microbiology and Epidemiology Academy of Military Medical Sciences 100850 Beijing China

Gene transfer into primary rat hepatocytes was performed by employing cationic liposome as DNA carrier and the specific ligand of hepatic asialoglycoprotein receptor (ASGPR), asialofetuin, as liver-targeting ligand. The results showed that asialofetuin, when added to the gene transfer complexes, could significantly increase the hepatocyte transfection efficiency, and alleviate the cellular toxicity of Lipofectin. Several synthetic ligands of ASGPR (galactosyl albumin) could also increase the transfection efficiency of hepatocyte like asialofetuin. It was proved that ASGPR and cationic liposome could synergistically mediate the gene transfer into primary rat hepatocytes. This novel gene delivery system provided a safer, more simple and efficient gene transfer method for primary hepatocytes, and showed prospecting application in hepatic gene therapy.

关键词： asialoglycoprotein receptor cationic liposome receptor-mediated gene transfer primary hepatocyte.

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Lactosamination of liposomes and hepatotropic targeting research

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World Journal of Gastroenterology 2000年第4期6卷 593-596页

作者： Chen YP Zhang L Lu QS Feng XR Luo KX First Mil Med Univ Dept Infect Dis Nanfang Hosp Guangzhou Guangdong Prov Peoples R China

Site-specific delivery of therapeutic drugs to their target cells is a major scientific challenge for the pharmaceutical sciences. It offers a number of advantages over conventional drug administration. With drug targeting, high local concentrations of the drug can be achieved, thus circumventing many unwanted side effects. Various carriers have been suggested for the delivery of drugs, including liposomes[1 - 5] and (neo ) glycoproteins[6-8]. The asialoglycoprotein receptor (ASGP-R) has frequently been utilized for targeting drugs to the parenchymal liver cell[6- 12]. Liposomes have several advantageous characteristics as drug carrier, and particularly, ligandtacked liposomes achieve a highly effective targeting[13]. Hara et al reported that asialofetuin (AF)-tacked liposomes distributed to rat hepatocytes selectively in vivo[14], and ASGP-R mediated the uptake of AF-liposomes encapsulating IFN-γ by isolated rat hepatocytes in vitro[15]. Lactosaminated human serum albumin (L-HSA) is a neoglycoprotein taking number of galactose residue as terminal sugar[6].

关键词： liposomes asialoglycoprotein liver interferon-alpha antigens viral drug carriers drug therapy rats

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Improved method increases sensitivity for circulating hepatocellular carcinoma cells

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World Journal of Gastroenterology 2015年第10期21卷 2918-2925页

作者： Hui-Ying Liu Hai-Hua Qian Xiao-Feng Zhang Jun Li Xia Yang Bin Sun Jun-Yong Ma Lei Chen Zheng-Feng Yin Molecular Oncology Laboratory Eastern Hepatobiliary Surgery HospitalSecond Military Medical University

AIM:To improve an asialoglycoprotein receptor(ASGPR)-based enrichment method for detection of circulating tumor cells(CTCs)of hepatocellular carcinoma(HCC).METHODS:Peripheral blood samples were collected from healthy subjects,patients with HCC or various other cancers,and patients with hepatic lesions or *** were enriched from whole blood by extracting CD45-expressing leukocytes with monoclonal antibody coated-beads following density gradient *** remaining cells were cytocentrifuged on polylysine-coated *** cells were treated by triple immunofluorescence staining with CD45antibody and a combination of antibodies against ASGPR and carbamoyl phosphate synthetase 1(CPS1),used as liver-specific markers,and costained with *** cell slide was imaged and stained tumor cells that met preset criteria were ***,sensitivity and specificity of the detection methods were determined and compared by spiking experiments with various types of cultured human tumor cell *** of ASGPR and CPS1 in cultured tumor cells and tumor tissue specimens was analyzed by flow cytometry and triple immunofluorescence staining,***:CD45 depletion of leukocytes resulted in a significantly greater recovery of multiple amounts of spiked HCC cells than the ASGPR+selection(P s<0.05).The expression rates of either ASGPR or CPS1were different in various liver cancer cell lines,ranging between 18%and 99%for ASGPR and between 9%and 98%for *** both human HCC tissues and liver cancer cell lines,there were a few HCC cells that did not stain positive for ASGPR or *** mixture of monoclonal antibodies against ASGPR and CPS1identified more HCC cells than either antibody ***,these antibodies did not detect any tumor cells in blood samples spiked with the human breastcancer cell line MCF-7 and the human renal cancer cell line ***+or/and CPS1+CTCs were detected in 29/32(91%)patients with HCC,but not in patients w

关键词： asialoglycoprotein receptor Carbamoyl phosphate sy

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Cellular fi bronectin stimulates hepatocytes to produce factors that promote alcohol-induced liver injury

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World Journal of Hepatology 2011年第2期3卷 45-55页

作者： Razia S Aziz-Seible Benita L McVicker Kusum K Kharbanda Carol A Casey Department of Biochemistry and Molecular Biology University of Nebraska Medical Center985870Nebraska Medical CenterOmahaNE68198-5870United States Liver Study Unit Omaha Veterans Affairs Medical Center4101 Woolworth AvenueOmahaNE68105United States

AIM:To examine the consequences of cellular f ibronectin(cFn)accumulation during alcohol-induced injury,and inv estigate whether increased cFn could have an effect on hepatocytes(HCs)by producing factors that could cont ribute to alcohol-induced liver ***:HCs were isolated from rats fed a control or ethanol liquid diet for four to six *** c Fn(up to 7.5 μg/mL)was added to cells cultured for 20 h,and viability(lactate dehydrogenase),apoptosis(caspase activity)and se cretion of proinflammat-ory cytokines(tumor ne c rosis fac tor alpha,TNF-α and interleukin 6,IL-6),mat rix metalloproteinases(MMPs)and their inhibitors(tissue inhibitors of metall-oproteinases,TIMPs)was det *** ation of iodinated cFn was det ermined over a 3 h time period in the ***:cFn degradation is impaired in HCs isolated from ethanol-fed animals,leading to its accumulation in the *** of exogenous cFn did not affect viability of HCs from control or ethanolfed animals,and apoptosis was affected only at the higher *** retion of MMPs,TIMPs,TNF-α and IL-6,however,was increased by exogenously added cFn,with HCs from ethanolfed animals showing increased susceptibility compared to the ***:These results suggest that the elevated amounts of cFn observed in alcoholic liver injury can stimulate hepatocytes to produce factors which promote further tissue damage.

关键词： Alcoholic liver diseases Hepatocytes Fibronectin asialoglycoprotein receptor Inflammation Fibrosis

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