AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transfor...AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.展开更多
摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-...摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-2012年11月在我院行冠脉造影检查确诊冠心病患者100例，并根据冠脉病变的支数分组，并人选同时期在我院行冠脉造影术明确无冠脉狭窄的非冠心病组（30例）。采用聚合酶链式反应（PCR）测定外周血单个核细胞中TIPE2的基因水平，采用双抗体夹心ELISA法测定血清中MCP-1和MMP-9水平。结果：①外周血单个核细胞TIPE2基因表达水平测定的结果：冠脉病变患者PBMC中TIPE2的基因表达水平明显低于非冠心病组，双支病变组低于单支病变组，多支病变组明显低于单支病变组和双支病变组，差异均具有统计学意义（P〈0．01）；②TIPE2基因表达水平与MCP-1、MMP-9水平呈负相关，相关系数分别为r=-0．560，r=-0．569，均P〈0．05）。结论：冠心病患者中TIPE2表达水平随冠脉病变血管支数的增加而降低，而且与MCP-1和MMP-9呈负相关。展开更多
目的探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中TIPE2(tumor necrosis factor-α induced protein 8 like 2)表达及其与SLE疾病活动度的关系。方法连续入选20例SLE患者做为SLE组,20名门诊健康体检者为对照组。采用实时荧光...目的探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中TIPE2(tumor necrosis factor-α induced protein 8 like 2)表达及其与SLE疾病活动度的关系。方法连续入选20例SLE患者做为SLE组,20名门诊健康体检者为对照组。采用实时荧光定量聚合酶链反应(RT-PCR),检测两组PBMCs中TIPE2mRNA的表达水平。以ΔCt=Ct(待测基因)-Ct(内参基因)来代表基因表达水平。结果 SLE患者PBMCsTIPE2mRNA表达水平(ΔCt=4.04±1.02)明显低于对照组(ΔCt=8.06±1.13,P<0.01);SLE患者PBMCs中TIPE2mRNA的表达水平与SLE疾病活动评分指数(SLEDAI)呈负相关(r=-0.568,P<0.01),而与SLE其他临床指标(如C3、C4、血沉等)无相关性。结论 SLE患者PBMCsTIPE2基因表达下降,且其表达水平与SLE疾病活动评分指数负相关,TIPE2基因可能在SLE的发病过程中发挥了一定作用。展开更多
文摘AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.
文摘摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-2012年11月在我院行冠脉造影检查确诊冠心病患者100例，并根据冠脉病变的支数分组，并人选同时期在我院行冠脉造影术明确无冠脉狭窄的非冠心病组（30例）。采用聚合酶链式反应（PCR）测定外周血单个核细胞中TIPE2的基因水平，采用双抗体夹心ELISA法测定血清中MCP-1和MMP-9水平。结果：①外周血单个核细胞TIPE2基因表达水平测定的结果：冠脉病变患者PBMC中TIPE2的基因表达水平明显低于非冠心病组，双支病变组低于单支病变组，多支病变组明显低于单支病变组和双支病变组，差异均具有统计学意义（P〈0．01）；②TIPE2基因表达水平与MCP-1、MMP-9水平呈负相关，相关系数分别为r=-0．560，r=-0．569，均P〈0．05）。结论：冠心病患者中TIPE2表达水平随冠脉病变血管支数的增加而降低，而且与MCP-1和MMP-9呈负相关。
文摘目的探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中TIPE2(tumor necrosis factor-α induced protein 8 like 2)表达及其与SLE疾病活动度的关系。方法连续入选20例SLE患者做为SLE组,20名门诊健康体检者为对照组。采用实时荧光定量聚合酶链反应(RT-PCR),检测两组PBMCs中TIPE2mRNA的表达水平。以ΔCt=Ct(待测基因)-Ct(内参基因)来代表基因表达水平。结果 SLE患者PBMCsTIPE2mRNA表达水平(ΔCt=4.04±1.02)明显低于对照组(ΔCt=8.06±1.13,P<0.01);SLE患者PBMCs中TIPE2mRNA的表达水平与SLE疾病活动评分指数(SLEDAI)呈负相关(r=-0.568,P<0.01),而与SLE其他临床指标(如C3、C4、血沉等)无相关性。结论 SLE患者PBMCsTIPE2基因表达下降,且其表达水平与SLE疾病活动评分指数负相关,TIPE2基因可能在SLE的发病过程中发挥了一定作用。
