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Proteomics to display tissue repair opposing injury response to LPS-induced liver injury

World Journal of Gastroenterology 2004年第18期10卷 2701-2705页

作者： Xiao-WeiLiu Fang-GenLu Guang-SenZhang Xiao-PingWu YuYou chun-huiouyang Dong-YeYang DepartmentofGastroenterology theSecondXiangyaHospitalCentralSouthUniversityChangsha410011HunanProvinceChina DepartmentofHematology theSecondXiangyaHospitalCentralSouthUniversityChangsha410011HunanProvinceChina

AIM: To examine the protein expression alterations in liver injury/repair network regulation as a response to gut-derived lipopolysaccharide (LPS) treatment, in order to anticipate the possible signal molecules or biomarkers in signaling LPS-related liver ***: Male BALB/c mice were treated with intraperitoneal (i.p.) LPS (4 mg/kg) and sacrificed at 0, 6, 24 and 30 h to obtain livers. The livers were stained with hematoxylin and eosin for histopathologic analyses. Total liver protein was separated by two-dimensional gel electrophoresis (2-DE). The peptide mass of liver injury or repair related proteins were drawn up and the protein database was searched to identify the ***: Observations were as follows: (1) TRAIL-R2 was down regulated in livers of LPS-treated mice. TNFAIP1 was significantly up regulated at 6 h, then down- regulated at 24, 30 h with silent expression during senescent stage.(2) The amount of metaxin 2 and mitochondria import inner membrane translocase subunit TIM8a (TIMM8A) was increased upon treatment with LPS, (3) P34 cdc2 kinase was significantly up-regulated 30 h after LPS administration with silent expression during senescent, 6, 24 h treated stage. (4) The amount of proteasome activator 28 alpha subunit (PA28), magnesium dependent protein phosphatase(MDPP) and lysophospholipase 2 was decreased 6 h after LPS treatment but recovered or up-regulated 24 and 30 h after LPS ***: LPS-treated mouse liver displaying a timedependent liver injury can result in expression change of some liver injury or repair related proteins.

关键词：组织修复免疫反应反作用 LPS-感应肝损伤脂多糖

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Study on relationship between expression level and molecular conformations of gene drugs targeting to hepatoma cells in vitro

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World Journal of Gastroenterology 2003年第9期9卷 1954-1958页

作者： Dong-YeYang Fang-GenLu Xi-XiangTang Shui-PingZhao chun-huiouyang Xiao-PingWu Xiao-WeiLiu Xiao-YingWu DepartmentofGastroenterology XiangyaSecondHospitalCentralSouthUniversityChangsha410011&InstituteofShengLifeGeneDrugsChangsha410003HunanChina DepartmentofCardiovascularology XiangyaSecondHospitalCentralSouthUniversityChangsha410011HunanChina DepartmentofElectronMicroscope XiangyaMedicalSchoolCentralSouthUniversityChangsha410011Hunan.China

AIM: To increase exogenous gene expression level bymodulating molecular conformations of targeting gene ***: The full length cDNAs of both P40 and P35subunits of human interleukin 12 were amplified throughpolymerase chain reaction (PCR) and cloned into eukaryoticexpressing vectors pcDNA3.1(±) to construct plasmids of P(+)/IL-12, P(+)/P40 and P(-)/P35. These plasmids werecombined with ASOR-PLL to form two targeting gene drugs[ASOR-PLL-P(+)/IL-12 and ASOR-PLL-P(+)/P40 + ASOR-PLL-P(-)/P35] in optimal ratios. The conformations of these twodrugs at various concentrations adjuvant were examined underelectron microscope (EM) and the drugs were transfected intoHepG2 (ASGr+) cells. Semi-quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) was performed withtotal RNA extracted from the transfected cells to determinethe hiL12 mRNA transcript level. The hiL12 protein in thecultured supernatant was measured with enzyme-linkedimmunosorbent assay (ELISA) 48 hours after ***: Targeting gene drugs, whose structures weregranular and circle-like and diameters ranged from 25 nmto 150 nm, had the highest hIL-12 expression level. ThehIL-12 expression level in the group co-transfected withASOR-PLL-P(+)/P4o and ASOR-PLL-P(-)/P35 was higher thanthat of ASOR-PLL-P(+)/IL-12 transfected ***: The molecular conformations of targetinggene drugs play an important role in exogenous geneexpression level, the best structures are granular and circle-like and their diameters range from 25 nm to 150 nm. Thesizes and linking styles of exogenous genes also have someeffects on their expression level.

关键词：肝癌基因疗法分子构造酶联免疫吸附试验

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