目的研究2型糖尿病大鼠肝脏肝细胞Tribbles同源蛋白(TRB)3的表达变化及丝胶的调节作用。方法选择36只SPF级雄性SD大鼠作为研究对象,随机分为正常对照组(12只)和造模组(24只)。造模组大鼠以高脂高糖饲料喂养+腹腔注射链脲佐菌素的方法构...目的研究2型糖尿病大鼠肝脏肝细胞Tribbles同源蛋白(TRB)3的表达变化及丝胶的调节作用。方法选择36只SPF级雄性SD大鼠作为研究对象,随机分为正常对照组(12只)和造模组(24只)。造模组大鼠以高脂高糖饲料喂养+腹腔注射链脲佐菌素的方法构建2型糖尿病大鼠模型,将成模的大鼠随机分为模型组和丝胶治疗组,每组12只,丝胶治疗组大鼠给予丝胶2.4 g/(kg·d)、其余组大鼠给予等体积生理盐水连续灌胃35 d。给药结束,取血液标本采用葡萄糖氧化酶法检测各组大鼠的空腹血糖;取肝脏组织采用SP免疫组织化学法、免疫印迹法和Real Time Q-PCR法分别检测各组大鼠肝细胞TRB3蛋白和mRNA的表达。结果与模型组大鼠相比,丝胶治疗组大鼠的空腹血糖水平、肝细胞TRB3蛋白和mRNA的表达明显降低(tSP免疫组织化学法=-2.32,t免疫印迹法=-4.89,t实时定量PCR法=-3.08,P<0.05)。结论丝胶降低血糖的作用,可能通过下调肝脏TRB3的表达进而降低对胰岛素磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路中Akt的抑制作用来实现。展开更多
AIM： To evaluate the possible role of Tribble 3 （TRB3） in a rat model of non-alcoholic fatty liver disease （NAFLD） and its signal transduction mechanism.METHODS： Thirty Sprague-Dawley rats were randomized into t...AIM： To evaluate the possible role of Tribble 3 （TRB3） in a rat model of non-alcoholic fatty liver disease （NAFLD） and its signal transduction mechanism.METHODS： Thirty Sprague-Dawley rats were randomized into three groups： normal control group, non-alcoholic fatty liver group A （fed on a high-fat diet for 8 wk） and group B （fed on a high-fat diet for 16 wk）. To determine the degree of hepatic steatosis in rats of each group, livers were stained with hematoxylin and eosin, and evaluated; real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction was performed to measure the expression levels of TRI33 mRNA, and Western blotting analysis was done to determine the expression levels of protein kinase B （Akt） and phosphorylated protein kinase B （p-Akt-Thr308, p-Akt-Ser473）.RESULTS： Hepatic steatosis was evident in both NAFLD groups： mild to moderate hepatic steatosis occurred in group A, mainly as mild steatosis.Moderate to severe hepatic steatosis occurred in group B, mainly as severe steatosis. The expression level of TRB3 mRNA in group B was significantly higher than in the control group （122.28 ± 95.37 vs 3.06 ± 2.33,P = 0.002） and group A （122.28 ± 95.37 vs 5.77 ± 4.20,P = 0.001）. There was no significant difference in the expression levels of Akt （1.03 ± 0.53 vs 1.12 ± 0.77,P = 0.729） and p-Akt-Thr308 （0.82 ± 0.45 vs 0.92 ± 0.38, P = 0.592） between group A and the control group. The expression level of Akt and p-Akt-Thr308 in group B was significantly lower than in group A （Akt 0.41 ± 0.16 vs 1.12 ± 0.77, P = 0.008; p-Akt-Thr308 0.47 ± 0.19 vs 0.82 ± 0.45, P = 0.036） and the control group （Akt 0.41 ± 0.16 vs 1.03 ± 0.53, P = 0.018;p-Akt-Thr308 0.47 ± 0.19 vs 0.92 ± 0.38, P = 0.010）.The expression level of p-Akt-Ser473 in group A was significantly higher than in group B （1.48 ± 0.50 vs 0.81± 0.39, P = 0.041） as well as the control group （1.48 ± 0.50 vs 0.45 ± 0.26, P = 0.003）.CONCLUSION： TRB3 blocks insulin signaling by inhibiting Akt activation, which contributes to insulin resistance. It may be an important factor in the occurrence and development of NAFLD.展开更多
目的:对Tribbles相关蛋白3(Tribbles related protein 3,TRB3)及CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein homologous protein,CHOP)在高脂高糖饮食诱导的大鼠非酒精性脂肪肝(nonalcoholic fatty liver disease,NA...目的:对Tribbles相关蛋白3(Tribbles related protein 3,TRB3)及CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein homologous protein,CHOP)在高脂高糖饮食诱导的大鼠非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)中的表达变化进行研究并探讨他们在NAFLD中的作用.方法:将30只Wistar大鼠随机分为正常组和NAFLD模型组,每组各15只.模型组大鼠采用高脂高糖饮食诱导NAFLD,正常组大鼠则给予普通饲料喂养,造模时间总计为16 wk.血清中总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白(highdensity lipoprotein,HDL)和低密度脂蛋白(lowdensity lipoprotein,LDL)的含量采用全自动生化分析仪进行检测;应用PCR技术对肝脏中TRB3及CHOP m RNA水平的改变进行检测;应用免疫组织化学技术对肝脏中TRB3及CHOP蛋白水平的改变进行检测;采用流式细胞仪检测细胞凋亡改变.结果:模型组大鼠血清中TC、TG和LDL含量较正常组大鼠显著升高(P<0.05),而HDL则明显低于正常组(P<0.05);与正常组相比,模型组大鼠肝脏中TRB3及CHOP m RNA水平均显著增加(P<0.05或P<0.01);免疫组织化学结果显示模型组大鼠肝脏中TRB3及CHOP蛋白表达水平较正常组大鼠显著升高(P<0.01);此外流式细胞仪对大鼠肝细胞凋亡进行检测发现,模型组大鼠肝细胞凋亡与正常组相比明显增多.结论:TRB3和CHOP在基因及蛋白水平表达上调可能与高脂高糖诱导的NAFLD的发生发展有关.展开更多
Objective Vascular endothelial growth factor A(VEGFA)is a key regulator of angiogenesis,which is a hallmark of cancer that promotes cancer growth and metastasis.It is of great significance to find new intervention tar...Objective Vascular endothelial growth factor A(VEGFA)is a key regulator of angiogenesis,which is a hallmark of cancer that promotes cancer growth and metastasis.It is of great significance to find new intervention targets and related regulatory mechanisms of VEGFA related angiogenesis for the treatment of tumors.This study focuses on the role of tribbles pseudokinase 3(TRIB3)/signal transducer and activator of transcription 3(STAT3)/VEGFA signaling axis in colon cancer angiogenesis.Methods This study investigated the expression level of TRIB3 in colon cancer through database analysis and tissue microarray analysis.The effect of TRIB3 on proliferation,migration and tube formation ability of human umbilical vein endothelial cells(HUVECs)was further confirmed by CCK8 assay,scratch-wound assay/migration assay and tube formation assay respectively.The regulatory relationship of TRIB3/VEGFA signaling axis was identified by qPCR and Western blotting,which was further confirmed through animal experiments,and the specific regulatory mechanism was explored by immunoprecipitation(IP)and chromatin immunoprecipitation(ChIP)with colon cancer cell lines.Results TRIB3 was increased in colon cancer tissues compared to normal tissues,and elevated TRIB3 expression indicated a poor prognosis in colon cancer patients.Moreover,it was found that silencing TRIB3 could inhibit cancer angiogenesis,whereas overexpressing TRIB3 promoted cancer angiogenesis in vitro and in vivo.Mechanistically,TRIB3 physically interacted with STAT3 and enhanced STAT3-mediated transcriptional activity.Furthermore,the function of TRIB3 in cancer angiogenesis was through cooperating with STAT3 to increase the VEGFA expression.Conclusion Our study provides insights into cancer angiogenesis and offers a potential therapeutic strategy for TRIB3-overexpressed cancer.展开更多
文摘目的研究2型糖尿病大鼠肝脏肝细胞Tribbles同源蛋白(TRB)3的表达变化及丝胶的调节作用。方法选择36只SPF级雄性SD大鼠作为研究对象,随机分为正常对照组(12只)和造模组(24只)。造模组大鼠以高脂高糖饲料喂养+腹腔注射链脲佐菌素的方法构建2型糖尿病大鼠模型,将成模的大鼠随机分为模型组和丝胶治疗组,每组12只,丝胶治疗组大鼠给予丝胶2.4 g/(kg·d)、其余组大鼠给予等体积生理盐水连续灌胃35 d。给药结束,取血液标本采用葡萄糖氧化酶法检测各组大鼠的空腹血糖;取肝脏组织采用SP免疫组织化学法、免疫印迹法和Real Time Q-PCR法分别检测各组大鼠肝细胞TRB3蛋白和mRNA的表达。结果与模型组大鼠相比,丝胶治疗组大鼠的空腹血糖水平、肝细胞TRB3蛋白和mRNA的表达明显降低(tSP免疫组织化学法=-2.32,t免疫印迹法=-4.89,t实时定量PCR法=-3.08,P<0.05)。结论丝胶降低血糖的作用,可能通过下调肝脏TRB3的表达进而降低对胰岛素磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路中Akt的抑制作用来实现。
文摘AIM： To evaluate the possible role of Tribble 3 （TRB3） in a rat model of non-alcoholic fatty liver disease （NAFLD） and its signal transduction mechanism.METHODS： Thirty Sprague-Dawley rats were randomized into three groups： normal control group, non-alcoholic fatty liver group A （fed on a high-fat diet for 8 wk） and group B （fed on a high-fat diet for 16 wk）. To determine the degree of hepatic steatosis in rats of each group, livers were stained with hematoxylin and eosin, and evaluated; real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction was performed to measure the expression levels of TRI33 mRNA, and Western blotting analysis was done to determine the expression levels of protein kinase B （Akt） and phosphorylated protein kinase B （p-Akt-Thr308, p-Akt-Ser473）.RESULTS： Hepatic steatosis was evident in both NAFLD groups： mild to moderate hepatic steatosis occurred in group A, mainly as mild steatosis.Moderate to severe hepatic steatosis occurred in group B, mainly as severe steatosis. The expression level of TRB3 mRNA in group B was significantly higher than in the control group （122.28 ± 95.37 vs 3.06 ± 2.33,P = 0.002） and group A （122.28 ± 95.37 vs 5.77 ± 4.20,P = 0.001）. There was no significant difference in the expression levels of Akt （1.03 ± 0.53 vs 1.12 ± 0.77,P = 0.729） and p-Akt-Thr308 （0.82 ± 0.45 vs 0.92 ± 0.38, P = 0.592） between group A and the control group. The expression level of Akt and p-Akt-Thr308 in group B was significantly lower than in group A （Akt 0.41 ± 0.16 vs 1.12 ± 0.77, P = 0.008; p-Akt-Thr308 0.47 ± 0.19 vs 0.82 ± 0.45, P = 0.036） and the control group （Akt 0.41 ± 0.16 vs 1.03 ± 0.53, P = 0.018;p-Akt-Thr308 0.47 ± 0.19 vs 0.92 ± 0.38, P = 0.010）.The expression level of p-Akt-Ser473 in group A was significantly higher than in group B （1.48 ± 0.50 vs 0.81± 0.39, P = 0.041） as well as the control group （1.48 ± 0.50 vs 0.45 ± 0.26, P = 0.003）.CONCLUSION： TRB3 blocks insulin signaling by inhibiting Akt activation, which contributes to insulin resistance. It may be an important factor in the occurrence and development of NAFLD.
文摘目的:对Tribbles相关蛋白3(Tribbles related protein 3,TRB3)及CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein homologous protein,CHOP)在高脂高糖饮食诱导的大鼠非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)中的表达变化进行研究并探讨他们在NAFLD中的作用.方法:将30只Wistar大鼠随机分为正常组和NAFLD模型组,每组各15只.模型组大鼠采用高脂高糖饮食诱导NAFLD,正常组大鼠则给予普通饲料喂养,造模时间总计为16 wk.血清中总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白(highdensity lipoprotein,HDL)和低密度脂蛋白(lowdensity lipoprotein,LDL)的含量采用全自动生化分析仪进行检测;应用PCR技术对肝脏中TRB3及CHOP m RNA水平的改变进行检测;应用免疫组织化学技术对肝脏中TRB3及CHOP蛋白水平的改变进行检测;采用流式细胞仪检测细胞凋亡改变.结果:模型组大鼠血清中TC、TG和LDL含量较正常组大鼠显著升高(P<0.05),而HDL则明显低于正常组(P<0.05);与正常组相比,模型组大鼠肝脏中TRB3及CHOP m RNA水平均显著增加(P<0.05或P<0.01);免疫组织化学结果显示模型组大鼠肝脏中TRB3及CHOP蛋白表达水平较正常组大鼠显著升高(P<0.01);此外流式细胞仪对大鼠肝细胞凋亡进行检测发现,模型组大鼠肝细胞凋亡与正常组相比明显增多.结论:TRB3和CHOP在基因及蛋白水平表达上调可能与高脂高糖诱导的NAFLD的发生发展有关.
基金supported by grants from the National Natural Science Foundation of China(No.81860249)Shihezi University Innovation and Development Project(No.CXFZ202113).
文摘Objective Vascular endothelial growth factor A(VEGFA)is a key regulator of angiogenesis,which is a hallmark of cancer that promotes cancer growth and metastasis.It is of great significance to find new intervention targets and related regulatory mechanisms of VEGFA related angiogenesis for the treatment of tumors.This study focuses on the role of tribbles pseudokinase 3(TRIB3)/signal transducer and activator of transcription 3(STAT3)/VEGFA signaling axis in colon cancer angiogenesis.Methods This study investigated the expression level of TRIB3 in colon cancer through database analysis and tissue microarray analysis.The effect of TRIB3 on proliferation,migration and tube formation ability of human umbilical vein endothelial cells(HUVECs)was further confirmed by CCK8 assay,scratch-wound assay/migration assay and tube formation assay respectively.The regulatory relationship of TRIB3/VEGFA signaling axis was identified by qPCR and Western blotting,which was further confirmed through animal experiments,and the specific regulatory mechanism was explored by immunoprecipitation(IP)and chromatin immunoprecipitation(ChIP)with colon cancer cell lines.Results TRIB3 was increased in colon cancer tissues compared to normal tissues,and elevated TRIB3 expression indicated a poor prognosis in colon cancer patients.Moreover,it was found that silencing TRIB3 could inhibit cancer angiogenesis,whereas overexpressing TRIB3 promoted cancer angiogenesis in vitro and in vivo.Mechanistically,TRIB3 physically interacted with STAT3 and enhanced STAT3-mediated transcriptional activity.Furthermore,the function of TRIB3 in cancer angiogenesis was through cooperating with STAT3 to increase the VEGFA expression.Conclusion Our study provides insights into cancer angiogenesis and offers a potential therapeutic strategy for TRIB3-overexpressed cancer.
