目的研究杞精明目汤药物血浆对结膜松弛症成纤维细胞肿瘤坏死因子α刺激基因6(TSG-6)和穿透素3(PTX3)表达的影响,探讨结膜松弛症的发病机制及杞精明目汤对其影响作用。方法制备杞精明目汤药物血浆和无药血浆,分别用含20%、10%、5%药物...目的研究杞精明目汤药物血浆对结膜松弛症成纤维细胞肿瘤坏死因子α刺激基因6(TSG-6)和穿透素3(PTX3)表达的影响,探讨结膜松弛症的发病机制及杞精明目汤对其影响作用。方法制备杞精明目汤药物血浆和无药血浆,分别用含20%、10%、5%药物血浆或无药血浆的细胞全培养基培养结膜松弛症成纤维细胞,4 h、24 h后分别提取m RNA和蛋白质,采用实时荧光定量PCR(real time PCR)技术和Western blot法检测细胞TSG-6、PTX3以及基质金属蛋白酶MMP-1、MMP-3的表达。结果 20%和10%的含药血浆均可有效上调结膜松弛症成纤维细胞TSG-6 m RNA和蛋白的表达,且作用接近(P>0.05),而5%浓度的含药血浆对TSG-6 m RNA和蛋白的表达无明显上调作用。三种浓度的含药血浆对结膜松弛症成纤维细胞PTX3 m RNA和蛋白的表达无明显作用。三种浓度的含药血浆均可明显抑制结膜松弛症成纤维细胞MMP-1 m RNA和蛋白的表达,其中20%和10%浓度对MMP-1 m RNA抑制作用较强,效果接近(P>0.05),其次为5%;20%浓度含药血浆对MMP-1蛋白表达的抑制作用最明显,10%和5%浓度对MMP-1蛋白表达的抑制作用无明显差异(P>0.05)。三种浓度的含药血浆均可有效抑制MMP-3 m RNA和蛋白的表达,其中对MMP-3 m RNA及蛋白的抑制作用最显著的均为20%浓度的含药血浆,其次为10%。结论 20%和10%浓度的杞精明目汤药物血浆能有效上调TSG-6的表达;20%、10%、5%三种浓度的含药血浆对PTX3的表达均无明显作用;三种浓度的杞精明目汤药物血浆均能有效抑制MMP-1、MMP-3的表达。展开更多
Background Pentraxin 3 （PTX3） is expressed in the heart under inflammatory conditions and plays an important role in atherogenesis. Patients with increased PTX3 levels may suffer from higher rates of cardiac events....Background Pentraxin 3 （PTX3） is expressed in the heart under inflammatory conditions and plays an important role in atherogenesis. Patients with increased PTX3 levels may suffer from higher rates of cardiac events. Regulation of specific genes by promoter methylation is important in atherogenesis. The factors influencing PTX3 levels and the association between epigenetics and PTX3 levels have not been investigated. Methods Blood samples were collected from 64 patients admitted to the Department of Cardiology, 35 who had coronary artery disease （CAD）, and 29 who were CAD-free. Plasma levels of PTX3 were measured by ELISA. PTX3 promoter methylation was evaluated via methyl-specific PCR. The severity of coronary artery lesion was evaluated by angiography. Results The level of PTX3 promoter methylation in the CAD group was 62.69% ± 20.57%, significantly lower than that of the CAD-free group, which was 72.45% ± 11.84% （P = 0.03）. Lower PTX3 promoter methylation levels in the CAD group were associated with higher plasma PTX3 concentrations （r = -0.29, P = 0.02）. Furthermore, lower PTX3 promoter methylation levels were associated with higher neutrophil to lymphocyte ratio （NLR） in men （r = -0.58, P = 0.002）. Conclusions The present study provides new evidence that methylation of the PTX3 promoter is associated with PTX3 plasma levels and NLR in coronary artery disease. This study also shows that modification of epigenetics by chronic inflamma- tion might be a significant molecular mechanism in the atherosclerotic processes that influence plasma PTX3 concentrations.展开更多
AIM:To identify the novel methylation-silenced gene pentraxin 3(PTX3) in esophageal squamous cell carcinoma(ESCC).METHODS:PTX3 mRNA expression was examined in six human ESCC cell lines,one human immortalized normal es...AIM:To identify the novel methylation-silenced gene pentraxin 3(PTX3) in esophageal squamous cell carcinoma(ESCC).METHODS:PTX3 mRNA expression was examined in six human ESCC cell lines,one human immortalized normal esophageal epithelial cell line,primary ESCC tumor tissue,and paired adjacent nontumor tissue using reverse transcription polymerase chain reaction(RTPCR).Semi-quantitative immunohistochemistry was used to examine cellular localisation and protein levels.Methylation specific PCR and bisulphite genomic sequencing were employed to investigate the methylation of the candidate gene.RESULTS:In the majority of ESCC cell lines,we found that PTX3 expression was down-regulated due to gene promoter hypermethylation,which was further confirmed by bisulphite genomic sequencing.Demethylation treatment with 5-aza-2'-deoxycytidine restored PTX3 mRNA expression in ESCC cell lines.Methylation was more common in tumor tissues(85%) than in adjacent nontumor tissues(25%)(P < 0.01).CONCLUSION:PTX3 is down-regulated through promoter hypermethylation in ESCC,and could potentially serve as a biomarker of ESCC.展开更多
Objective To determine whether the onset of acute lung injury （ALl） induces the up-regulation of pentraxin 3 （PTX3） expression in mice and whether PTX3 concentration in the biofluid can help recognizing sepsis-ind...Objective To determine whether the onset of acute lung injury （ALl） induces the up-regulation of pentraxin 3 （PTX3） expression in mice and whether PTX3 concentration in the biofluid can help recognizing sepsis-induced ALI. Methods Wild-type C57BL/6 mice （12-14 weeks old） were randomly divided into 3 groups. Mice in the group 1 （n=12） and group 2 （n=12） were instilled with lipopolysaccharide via intratracheal or intraperitoneal routes, respectively. Mice in the group 3 （n=8） were taken as blank controls. Pulmonary morphological and functional alterations were measured to determine the presence of experimental ALl. PTX3 expression in the lung was quantified at both protein and mRNA levels. PTX3 protein concentration in blood and bronchoalveolar lavage fluid was measured to evaluate its ability to diagnose sepsis-induced ALI by computing area under receiver operator characteristic curve （AUROCC）. Results ALl was commonly confirmed in the group 1 but never in the other groups. PTX3 expression was up-regulated indiscriminately among lipopolysaccharide-challenged mice. PTX3 protein concentration in the biofluid was unable to diagnose sepsis-induced ALl evidenced by its small AUROCC. PTX3 concentration in bronchoalveolar lavage fluid did not correlate with that in serum. Conclusions Lipopolysaccharide challenges induced PTX3 expression in mice regardless of the presence ofALI. PTX3 may act as an indicator of inflammatory response instead of organ injury per se.展开更多
AIM： To investigate the expression of pentraxin 3（PTX3） in rat corneal epithelium at the early stage of Aspergillus fumigatus（A. fumigatus） infection. METHODS： A total of 50 Wistar rats were randomly divided int...AIM： To investigate the expression of pentraxin 3（PTX3） in rat corneal epithelium at the early stage of Aspergillus fumigatus（A. fumigatus） infection. METHODS： A total of 50 Wistar rats were randomly divided into control group, Sham group and experimental group（fungal keratitis group, FK group）. The right eye was chosen as the experiment one and infected by A. fumigatus. Rats were executed at 8, 16 and 24 h after the experimental models being established. Corneal epithelia were collected to assess the expression of PTX3 by quantitative reverse transcription polymerase chain reaction（q RT-PCR） and Western blot analysis. RESULTS： Corneal inflammation scores increased as infection prolonged（P〈0.05, P〈0.001）. PTX3 m RNA expression was low in normal and Sham group rats＇ corneas. Level of PTX3 m RNA in infected rat cornea was elevated at 8 h and peaked at 16 h. The difference was significant compared with control group（P〈0.001）. Western blot analysis also showed a significant increase of PTX3 protein in experimental group at 8 h and peaked at 16 h（P〈0.001）. The synchronous expression of control group and experimental group were also in significant difference（P〈0.001）. CONCLUSION： PTX3 exists in cornea epithelium and is significantly increased after A. fumigatus infection. PTX3 plays an important role in the early stage of cornea innate immunity against A. fumigatus.展开更多
文摘目的研究杞精明目汤药物血浆对结膜松弛症成纤维细胞肿瘤坏死因子α刺激基因6(TSG-6)和穿透素3(PTX3)表达的影响,探讨结膜松弛症的发病机制及杞精明目汤对其影响作用。方法制备杞精明目汤药物血浆和无药血浆,分别用含20%、10%、5%药物血浆或无药血浆的细胞全培养基培养结膜松弛症成纤维细胞,4 h、24 h后分别提取m RNA和蛋白质,采用实时荧光定量PCR(real time PCR)技术和Western blot法检测细胞TSG-6、PTX3以及基质金属蛋白酶MMP-1、MMP-3的表达。结果 20%和10%的含药血浆均可有效上调结膜松弛症成纤维细胞TSG-6 m RNA和蛋白的表达,且作用接近(P>0.05),而5%浓度的含药血浆对TSG-6 m RNA和蛋白的表达无明显上调作用。三种浓度的含药血浆对结膜松弛症成纤维细胞PTX3 m RNA和蛋白的表达无明显作用。三种浓度的含药血浆均可明显抑制结膜松弛症成纤维细胞MMP-1 m RNA和蛋白的表达,其中20%和10%浓度对MMP-1 m RNA抑制作用较强,效果接近(P>0.05),其次为5%;20%浓度含药血浆对MMP-1蛋白表达的抑制作用最明显,10%和5%浓度对MMP-1蛋白表达的抑制作用无明显差异(P>0.05)。三种浓度的含药血浆均可有效抑制MMP-3 m RNA和蛋白的表达,其中对MMP-3 m RNA及蛋白的抑制作用最显著的均为20%浓度的含药血浆,其次为10%。结论 20%和10%浓度的杞精明目汤药物血浆能有效上调TSG-6的表达;20%、10%、5%三种浓度的含药血浆对PTX3的表达均无明显作用;三种浓度的杞精明目汤药物血浆均能有效抑制MMP-1、MMP-3的表达。
文摘Background Pentraxin 3 （PTX3） is expressed in the heart under inflammatory conditions and plays an important role in atherogenesis. Patients with increased PTX3 levels may suffer from higher rates of cardiac events. Regulation of specific genes by promoter methylation is important in atherogenesis. The factors influencing PTX3 levels and the association between epigenetics and PTX3 levels have not been investigated. Methods Blood samples were collected from 64 patients admitted to the Department of Cardiology, 35 who had coronary artery disease （CAD）, and 29 who were CAD-free. Plasma levels of PTX3 were measured by ELISA. PTX3 promoter methylation was evaluated via methyl-specific PCR. The severity of coronary artery lesion was evaluated by angiography. Results The level of PTX3 promoter methylation in the CAD group was 62.69% ± 20.57%, significantly lower than that of the CAD-free group, which was 72.45% ± 11.84% （P = 0.03）. Lower PTX3 promoter methylation levels in the CAD group were associated with higher plasma PTX3 concentrations （r = -0.29, P = 0.02）. Furthermore, lower PTX3 promoter methylation levels were associated with higher neutrophil to lymphocyte ratio （NLR） in men （r = -0.58, P = 0.002）. Conclusions The present study provides new evidence that methylation of the PTX3 promoter is associated with PTX3 plasma levels and NLR in coronary artery disease. This study also shows that modification of epigenetics by chronic inflamma- tion might be a significant molecular mechanism in the atherosclerotic processes that influence plasma PTX3 concentrations.
基金Supported by National High Technology Research and Development Program of China (863 Program),No. 2007AA02Z4Z4China Postdoctoral Science Foundation,No. 20090460394Beijing Municipal Natural Science Foundation,No. 7072022
文摘AIM:To identify the novel methylation-silenced gene pentraxin 3(PTX3) in esophageal squamous cell carcinoma(ESCC).METHODS:PTX3 mRNA expression was examined in six human ESCC cell lines,one human immortalized normal esophageal epithelial cell line,primary ESCC tumor tissue,and paired adjacent nontumor tissue using reverse transcription polymerase chain reaction(RTPCR).Semi-quantitative immunohistochemistry was used to examine cellular localisation and protein levels.Methylation specific PCR and bisulphite genomic sequencing were employed to investigate the methylation of the candidate gene.RESULTS:In the majority of ESCC cell lines,we found that PTX3 expression was down-regulated due to gene promoter hypermethylation,which was further confirmed by bisulphite genomic sequencing.Demethylation treatment with 5-aza-2'-deoxycytidine restored PTX3 mRNA expression in ESCC cell lines.Methylation was more common in tumor tissues(85%) than in adjacent nontumor tissues(25%)(P < 0.01).CONCLUSION:PTX3 is down-regulated through promoter hypermethylation in ESCC,and could potentially serve as a biomarker of ESCC.
基金Partly supported by a grant from Jie-shou Li Academician Gut Barrier Research Fund
文摘Objective To determine whether the onset of acute lung injury （ALl） induces the up-regulation of pentraxin 3 （PTX3） expression in mice and whether PTX3 concentration in the biofluid can help recognizing sepsis-induced ALI. Methods Wild-type C57BL/6 mice （12-14 weeks old） were randomly divided into 3 groups. Mice in the group 1 （n=12） and group 2 （n=12） were instilled with lipopolysaccharide via intratracheal or intraperitoneal routes, respectively. Mice in the group 3 （n=8） were taken as blank controls. Pulmonary morphological and functional alterations were measured to determine the presence of experimental ALl. PTX3 expression in the lung was quantified at both protein and mRNA levels. PTX3 protein concentration in blood and bronchoalveolar lavage fluid was measured to evaluate its ability to diagnose sepsis-induced ALI by computing area under receiver operator characteristic curve （AUROCC）. Results ALl was commonly confirmed in the group 1 but never in the other groups. PTX3 expression was up-regulated indiscriminately among lipopolysaccharide-challenged mice. PTX3 protein concentration in the biofluid was unable to diagnose sepsis-induced ALl evidenced by its small AUROCC. PTX3 concentration in bronchoalveolar lavage fluid did not correlate with that in serum. Conclusions Lipopolysaccharide challenges induced PTX3 expression in mice regardless of the presence ofALI. PTX3 may act as an indicator of inflammatory response instead of organ injury per se.
基金Supported by National Natural Science Foundation of China (No.81170825 No.81470609)+2 种基金Specialized Research Fund for the Doctoral Program of Higher Education (No.20123706110003)the Youth Natural Science Foundation of Shandong Province (No.ZR2013HQ007)the Key Project of Natural Science Foundation of Shandong Province (No.ZR2012HZ001).
文摘AIM： To investigate the expression of pentraxin 3（PTX3） in rat corneal epithelium at the early stage of Aspergillus fumigatus（A. fumigatus） infection. METHODS： A total of 50 Wistar rats were randomly divided into control group, Sham group and experimental group（fungal keratitis group, FK group）. The right eye was chosen as the experiment one and infected by A. fumigatus. Rats were executed at 8, 16 and 24 h after the experimental models being established. Corneal epithelia were collected to assess the expression of PTX3 by quantitative reverse transcription polymerase chain reaction（q RT-PCR） and Western blot analysis. RESULTS： Corneal inflammation scores increased as infection prolonged（P〈0.05, P〈0.001）. PTX3 m RNA expression was low in normal and Sham group rats＇ corneas. Level of PTX3 m RNA in infected rat cornea was elevated at 8 h and peaked at 16 h. The difference was significant compared with control group（P〈0.001）. Western blot analysis also showed a significant increase of PTX3 protein in experimental group at 8 h and peaked at 16 h（P〈0.001）. The synchronous expression of control group and experimental group were also in significant difference（P〈0.001）. CONCLUSION： PTX3 exists in cornea epithelium and is significantly increased after A. fumigatus infection. PTX3 plays an important role in the early stage of cornea innate immunity against A. fumigatus.
