本试验旨在考察牛膝多糖(ABPS)对脂多糖(LPS)免疫应激下仔猪空肠上皮细胞(IPEC-J2)促炎细胞因子分泌和表达的影响,并探讨ABPS调控IPEC-J2免疫应激可能的作用机制。选用4~5代的IPEC-J2,培养基中分别添加0(对照)、300、600、900、1 200μg...本试验旨在考察牛膝多糖(ABPS)对脂多糖(LPS)免疫应激下仔猪空肠上皮细胞(IPEC-J2)促炎细胞因子分泌和表达的影响,并探讨ABPS调控IPEC-J2免疫应激可能的作用机制。选用4~5代的IPEC-J2,培养基中分别添加0(对照)、300、600、900、1 200μg/m L ABPS和10μg/m L LPS,每组12个重复,每孔为1个重复。培养72 h后,采用酶联免疫吸附测定(ELISA)方法检测ABPS对促炎细胞因子白细胞介素1(IL-1)、白细胞介素6(IL-6)、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)分泌量的影响,采用实时定量PCR测定Toll样受体4(TLR4)、核转录因子κB(NF-κB)的mRNA表达量,采用Western blot法测定TLR4、NF-κB、磷酸化核转录因子κB(p-NF-κB)蛋白表达量。结果显示:与对照组相比,300、600、900和1 200μg/m L ABPS组能显著减少IL-1、IL-6、IL-8和TNF-α的分泌量(P<0.05);300μg/m L ABPS组能显著减少p-NF-κB蛋白的表达量(P<0.05),900和1 200μg/m L ABPS组能显著减少TLR4、NF-κB的mRNA和NF-κB蛋白的表达量(P<0.05)。由此可见,ABPS通过TLR4/NF-κB信号转导途径来调控促炎细胞因子的分泌,从而缓解免疫应激,低浓度ABPS通过直接抑制NF-κB磷酸化过程来降低免疫应激,高浓度ABPS则是通过抑制TLR4 mRNA、NF-κB mRNA和NF-κB蛋白的表达量来缓解免疫应激。展开更多
目的:探讨高迁移率族蛋白B1(high mobility group protein B1,HMGB1)和Toll样受体4(Toll-like receptor 4,TLR4)在人食管鳞状细胞癌(食管鳞癌)组织中的表达及其临床意义.方法:选择72例食管鳞癌标本,15例癌旁正常组织标本,采用En Vision...目的:探讨高迁移率族蛋白B1(high mobility group protein B1,HMGB1)和Toll样受体4(Toll-like receptor 4,TLR4)在人食管鳞状细胞癌(食管鳞癌)组织中的表达及其临床意义.方法:选择72例食管鳞癌标本,15例癌旁正常组织标本,采用En Vision免疫组织化学染色法检测HMGB1和TLR4在食管鳞癌组织及癌旁正常组织中的表达,并应用统计学方法对其表达与临床病理因素进行分析.结果:食管鳞癌组织中HMGB1、TLR4的表达显著高于正常组织(P<0.05),且与淋巴结转移及TNM分期相关(P<0.05),与肿瘤大小、分化程度等无相关性.食管麟癌组织HMGB1和TLR4的表达呈显著正相关(r=0.377,P<0.01).结论:食管鳞癌组织中HMGB1、TLR4的表达显著高于癌旁正常组织,且其表达与淋巴结转移及TNM分期相关,联合检测二者可能有助于评估食管鳞癌的恶性程度.因此,HMGB1/TLR信号通路有可能作为反映食管癌预后的重要生物学指标及抗食管癌的重要靶点.展开更多
AIM:To determine if the presence H pylori or its viru- lence affect toll-like receptor 4 (TLR4) and TLR5 mRNA expression levels. METHODS:For the in vivo assays, gastric biopsies were obtained from 40 patients and H py...AIM:To determine if the presence H pylori or its viru- lence affect toll-like receptor 4 (TLR4) and TLR5 mRNA expression levels. METHODS:For the in vivo assays, gastric biopsies were obtained from 40 patients and H pylori status was determined. For the in vitro assays, human gastric adenocarcinoma mucosal cells (AGS) were cultured in the presence or absence of twelve selected H pylori strains. H pylori strains isolated from culture-positive patients and selected strains were genotyped for cagA and vacA. The cDNA was obtained from mRNA extracted from biopsies and from infected AGS cells. TLR4 and TLR5 mRNA levels were examined by real-time PCR. RESULTS: The presence of H pylori did not affect the mRNA levels of TLR4 or TLR5 in gastric biopsies. The mRNA levels of both receptors were not influenced by the vacA status (P > 0.05 for both receptors) andthere were no differences in TLR4 or TLR5 mRNA levels among the different clinical presentations/histological fi ndings (P > 0.05). In the in vitro assay, the mRNA levels of TLR4 or TLR5 in AGS cells were not influenced by the vacAs1 status or the clinical condition as-sociated with the strains (P > 0.05 for both TLR4 and TLR5). CONCLUSION: The results of this study show that the mRNA levels of TLR4 and TLR5 in gastric cells, both in vivo and in vitro, are independent of H pylori colonization and suggest that vacA may not be a significant player in the first step of innate immune recognition mediated by TLR4 or TLR5.展开更多
文摘背景与目的:有研究证实,正丁酸钠(sodium butyrate,Na B)对肿瘤细胞具有抑制增殖、诱导分化和促凋亡的作用,但对涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC)的作用机制尚不明确。该研究主要探讨不同浓度Na B对SACC细胞株ACC-M侵袭、迁移的影响及其作用机制。方法:MTT法探索Na B作用ACC-M细胞的最佳浓度并观察细胞的生长情况,Transwell小室实验检测Na B对ACC-M细胞侵袭、迁移能力的影响,实时荧光定量PCR(real-time PCR,RT-PCR)和蛋白质印迹法(Western blot)分别检测5组浓度药物作用后ACC-M细胞中高迁移率蛋白-1(high mobility group box-1,HMGB1)、toll样受体-4(toll like receptor-4,TLR4)m RNA和蛋白的表达。结果:与对照组相比,加入Na B 0.625、1.25、2.5、5及10 mmol/L的5组均能抑制ACC-M细胞增殖且呈明显浓度依赖性(P<0.05)。5组浓度Na B均可抑制ACC-M细胞体外侵袭和迁移能力(P<0.05),同时能降低ACC-M细胞HMGB1、TLR4 m RNA及蛋白的表达(P<0.05);相关性分析显示TLR4蛋白表达的降低与HMGB1的抑制呈正相关(r=0.810,P<0.05)。结论:Na B可抑制ACC-M细胞增殖,显著降低ACC-M细胞的侵袭和迁移能力,同时降低HMGB1、TLR4 m RNA和蛋白的表达,且2者表达量呈明显正相关,提示Na B可能是通过下调HMGB1、TLR4 m RNA和蛋白的表达来实现对ACC-M细胞侵袭、迁移能力的抑制。
文摘本试验旨在考察牛膝多糖(ABPS)对脂多糖(LPS)免疫应激下仔猪空肠上皮细胞(IPEC-J2)促炎细胞因子分泌和表达的影响,并探讨ABPS调控IPEC-J2免疫应激可能的作用机制。选用4~5代的IPEC-J2,培养基中分别添加0(对照)、300、600、900、1 200μg/m L ABPS和10μg/m L LPS,每组12个重复,每孔为1个重复。培养72 h后,采用酶联免疫吸附测定(ELISA)方法检测ABPS对促炎细胞因子白细胞介素1(IL-1)、白细胞介素6(IL-6)、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)分泌量的影响,采用实时定量PCR测定Toll样受体4(TLR4)、核转录因子κB(NF-κB)的mRNA表达量,采用Western blot法测定TLR4、NF-κB、磷酸化核转录因子κB(p-NF-κB)蛋白表达量。结果显示:与对照组相比,300、600、900和1 200μg/m L ABPS组能显著减少IL-1、IL-6、IL-8和TNF-α的分泌量(P<0.05);300μg/m L ABPS组能显著减少p-NF-κB蛋白的表达量(P<0.05),900和1 200μg/m L ABPS组能显著减少TLR4、NF-κB的mRNA和NF-κB蛋白的表达量(P<0.05)。由此可见,ABPS通过TLR4/NF-κB信号转导途径来调控促炎细胞因子的分泌,从而缓解免疫应激,低浓度ABPS通过直接抑制NF-κB磷酸化过程来降低免疫应激,高浓度ABPS则是通过抑制TLR4 mRNA、NF-κB mRNA和NF-κB蛋白的表达量来缓解免疫应激。
文摘目的:探讨高迁移率族蛋白B1(high mobility group protein B1,HMGB1)和Toll样受体4(Toll-like receptor 4,TLR4)在人食管鳞状细胞癌(食管鳞癌)组织中的表达及其临床意义.方法:选择72例食管鳞癌标本,15例癌旁正常组织标本,采用En Vision免疫组织化学染色法检测HMGB1和TLR4在食管鳞癌组织及癌旁正常组织中的表达,并应用统计学方法对其表达与临床病理因素进行分析.结果:食管鳞癌组织中HMGB1、TLR4的表达显著高于正常组织(P<0.05),且与淋巴结转移及TNM分期相关(P<0.05),与肿瘤大小、分化程度等无相关性.食管麟癌组织HMGB1和TLR4的表达呈显著正相关(r=0.377,P<0.01).结论:食管鳞癌组织中HMGB1、TLR4的表达显著高于癌旁正常组织,且其表达与淋巴结转移及TNM分期相关,联合检测二者可能有助于评估食管鳞癌的恶性程度.因此,HMGB1/TLR信号通路有可能作为反映食管癌预后的重要生物学指标及抗食管癌的重要靶点.
文摘AIM:To determine if the presence H pylori or its viru- lence affect toll-like receptor 4 (TLR4) and TLR5 mRNA expression levels. METHODS:For the in vivo assays, gastric biopsies were obtained from 40 patients and H pylori status was determined. For the in vitro assays, human gastric adenocarcinoma mucosal cells (AGS) were cultured in the presence or absence of twelve selected H pylori strains. H pylori strains isolated from culture-positive patients and selected strains were genotyped for cagA and vacA. The cDNA was obtained from mRNA extracted from biopsies and from infected AGS cells. TLR4 and TLR5 mRNA levels were examined by real-time PCR. RESULTS: The presence of H pylori did not affect the mRNA levels of TLR4 or TLR5 in gastric biopsies. The mRNA levels of both receptors were not influenced by the vacA status (P > 0.05 for both receptors) andthere were no differences in TLR4 or TLR5 mRNA levels among the different clinical presentations/histological fi ndings (P > 0.05). In the in vitro assay, the mRNA levels of TLR4 or TLR5 in AGS cells were not influenced by the vacAs1 status or the clinical condition as-sociated with the strains (P > 0.05 for both TLR4 and TLR5). CONCLUSION: The results of this study show that the mRNA levels of TLR4 and TLR5 in gastric cells, both in vivo and in vitro, are independent of H pylori colonization and suggest that vacA may not be a significant player in the first step of innate immune recognition mediated by TLR4 or TLR5.
