目的检测乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)对人血管生成因子bFGF基因转录的激活作用并确定其作用于bFGF基因启动子的区域,分析HBx激活bFGF基因转录可能的分子机制,为揭示HBx促进肝癌发生的分子机制及发现新的治疗靶...目的检测乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)对人血管生成因子bFGF基因转录的激活作用并确定其作用于bFGF基因启动子的区域,分析HBx激活bFGF基因转录可能的分子机制,为揭示HBx促进肝癌发生的分子机制及发现新的治疗靶点。方法运用高保真DNA聚合酶从人肝癌HepG2细胞cDNA文库中扩增出bFGF基因启动子DNA序列;从含完整HBV DNA序列的表达载体中扩增HBx编码序列并克隆入真核表达载体p CDNA3-FLAG中,将HBx表达载体转染人肝癌HepG2细胞中,裂解细胞进行蛋白SDS-PAGE电泳,Western blot检测HBx蛋白的表达;将HBx表达载体和含bFGF基因启动子不同区段的荧光素酶报告载体共转染肝癌HepG2和SMMC-7721细胞,检测HBx对bFGF基因启动子转录活性的影响;运用Western blot检测在肝癌细胞中HBx过表达对ERK磷酸化的影响。结果成功克隆了HBx并使其在肝癌细胞中得到表达;成功克隆了bFGF基因启动子2.2 kb、1.4 kb、720 bp和360 bp的区段;在肝癌细胞SMMC-7721和HepG2细胞中HBx的表达能升高bFGF基因启动子的活性,且对上述4个区段的活性均增加约5倍;HBx以剂量依赖的方式增强bFGF基因启动子的活性;HBx高表达在肝癌HepG2细胞中不能升高ERK1/2的磷酸化水平。结论HBx能够激活bFGF基因启动子的活性,这种激活作用位于HBx启动子转录起始位点上游360 bp片段内,HBx促进bFGF基因转录不依赖于ERK1/2的磷酸化作用,提示HBx可能通过其它信号通路作用于bFGF基因的转录过程。展开更多
Objective:This study was conducted to evaluate the efficacy of Astragalus on diabetic foot,as well as the effects on the levels of serum VEGF,bFGF,MMP-2,and inflammatory factors in patients,and to provide a scientific...Objective:This study was conducted to evaluate the efficacy of Astragalus on diabetic foot,as well as the effects on the levels of serum VEGF,bFGF,MMP-2,and inflammatory factors in patients,and to provide a scientific basis for the treatment of diabetic foot with the traditional Chinese medicine Astragalus.Methods:By taking 100 cases of diabetic foot patients who were admitted to the metabolic internal medicine division of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine and met the criteria of natriuresis during April 2021-April 2023 as the study subjects,and according to the random number method,all patients were divided into the control group and the observation group,with 50 cases in each group.In the control group,only basic treatment was carried out,while in the observation group,Astragalus injection was added based on the control group.After 8 weeks of treatment,the clinical efficacy,serum VEGF,bFGF,MMP-2,and inflammatory factor levels of the patients in the two groups were compared,respectively.Results:The total clinical efficiency of patients in the observation group was significantly better than that in the control group(χ^(2)=5.01,P<0.05).The inflammatory factor indexes decreased substantially in both groups.However,the decrease in the observation group was significantly higher than in the control group(P<0.05).Compared with the control group,serum VEGF and bFGF were considerably higher in the observation group,while MMP-2 was significantly lower(P<0.05).Conclusion:Astragali Radix is clinically effective in the diabetic foot,which can induce vascular endothelial repair and reduce the level of inflammatory factors,to improve the inflammatory state of patients and promote the restoration of ulcerated wound tissue,which is worth promoting in clinical practice.展开更多
文摘Objective:This study was conducted to evaluate the efficacy of Astragalus on diabetic foot,as well as the effects on the levels of serum VEGF,bFGF,MMP-2,and inflammatory factors in patients,and to provide a scientific basis for the treatment of diabetic foot with the traditional Chinese medicine Astragalus.Methods:By taking 100 cases of diabetic foot patients who were admitted to the metabolic internal medicine division of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine and met the criteria of natriuresis during April 2021-April 2023 as the study subjects,and according to the random number method,all patients were divided into the control group and the observation group,with 50 cases in each group.In the control group,only basic treatment was carried out,while in the observation group,Astragalus injection was added based on the control group.After 8 weeks of treatment,the clinical efficacy,serum VEGF,bFGF,MMP-2,and inflammatory factor levels of the patients in the two groups were compared,respectively.Results:The total clinical efficiency of patients in the observation group was significantly better than that in the control group(χ^(2)=5.01,P<0.05).The inflammatory factor indexes decreased substantially in both groups.However,the decrease in the observation group was significantly higher than in the control group(P<0.05).Compared with the control group,serum VEGF and bFGF were considerably higher in the observation group,while MMP-2 was significantly lower(P<0.05).Conclusion:Astragali Radix is clinically effective in the diabetic foot,which can induce vascular endothelial repair and reduce the level of inflammatory factors,to improve the inflammatory state of patients and promote the restoration of ulcerated wound tissue,which is worth promoting in clinical practice.
