目的探究lncRNA GClnc1是否通过SIRT3/FOXO3/SOD2调控胃癌细胞增殖和迁移。方法通过转染慢病毒的方法,在人胃癌细胞SGC7901中分别过表达和沉默GClnc1,采用Western blot和qRT-PCR检测SIRT3/FOXO3/SOD2 mRNA和蛋白水平的改变;在过表达GCl...目的探究lncRNA GClnc1是否通过SIRT3/FOXO3/SOD2调控胃癌细胞增殖和迁移。方法通过转染慢病毒的方法,在人胃癌细胞SGC7901中分别过表达和沉默GClnc1,采用Western blot和qRT-PCR检测SIRT3/FOXO3/SOD2 mRNA和蛋白水平的改变;在过表达GClnc1的同时分别沉默SIRT3/FOXO3/SOD2,采用CCK-8检测细胞增殖能力,划痕实验检测细胞迁移能力。结果过表达GClnc1可以上调SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并增强SGC7901细胞的增殖[(1.00±0.14)vs(1.79±0.21),P<0.05]和迁移能力(10.87±0.76 vs 16.53±2.25,P<0.05),而沉默GClnc1则降低SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并减少细胞增殖(1.00±0.14 vs 0.62±0.13,P<0.05)和迁移能力(10.87±0.76 vs 6.46±0.89,P<0.01)。沉默SIRT3/FOXO3/SOD2可以消除过表达GClnc1对细胞增殖和迁移能力的促进作用。结论 GClnc1通过激活SIRT3/FOXO3/SOD2信号通路促进SGC7901细胞增殖和迁移。展开更多
目的研究微小RNA(microRNA,miRNA)miR-25-3p对心肌成纤维细胞纤维化表型的调控及机制。方法血管紧张素Ⅱ(AngⅡ)灌注小鼠构建心肌纤维化模型。用miRNA表达谱芯片检测纤维化的小鼠心肌中表达水平有差异的miRNA。原代分离法获得C57BL/6小...目的研究微小RNA(microRNA,miRNA)miR-25-3p对心肌成纤维细胞纤维化表型的调控及机制。方法血管紧张素Ⅱ(AngⅡ)灌注小鼠构建心肌纤维化模型。用miRNA表达谱芯片检测纤维化的小鼠心肌中表达水平有差异的miRNA。原代分离法获得C57BL/6小鼠心肌成纤维细胞(mCF),建立AngⅡ处理mCF的心肌纤维化细胞模型。mCF转染miR-25-3p mimic后检测纤维化相关基因的表达。双荧光素酶报告基因实验验证miR-25-3p与B细胞易位基因2(BTG2)的3′端非翻译区(3′UTR)的结合作用。放线菌素D实验验证BTG2对超氧化物歧化酶2(SOD2)稳定性的作用。结果miR-25-3p在AngⅡ灌注诱导的小鼠心肌和AngⅡ处理的mCF中表达升高。转染miR-25-3p mimic可使mCF中的Ⅰ型胶原α1(COL1A1)、Ⅲ型胶原α1(COL3A1)和α-平滑肌肌动蛋白(α-SMA)基因表达显著增加。双荧光素酶报告基因实验和功能实验证实BTG2是miR-25-3p的下游靶基因,并参与介导miR-25-3p促进mCF中纤维化相关基因表达的作用。机制研究证实BTG2可通过增加SOD2 m RNA的稳定性而上调其表达。结论miR-25-3p通过抑制BTG2的表达来下调SOD2的水平进而增强纤维化相关基因的表达,发挥促进心肌纤维化的作用。展开更多
Objective: To explore the effects of myocardial SOD2 by strenuous endurance exercise. Methods: 27 grown male SD rats were randomly divided into control group (C), a single bout of strenuous endurance exercise group (E...Objective: To explore the effects of myocardial SOD2 by strenuous endurance exercise. Methods: 27 grown male SD rats were randomly divided into control group (C), a single bout of strenuous endurance exercise group (E1) and seventh-week strenuous endurance exercise group (E2). Real-time PCR was used to observe the changes of mRNA expression for myocardial SOD2. Western bolt was used to observe the changes of SOD2 protein expression. In addition, SOD2, T-SOD and SOD1 activity changes were observed. Results: Myocardial SOD2 expression level at mRNA and protein of Group E1, E2 was significantly higher than that in group C, and SOD2 and T-SOD activity in group E2 were significantly higher than those in group C. Those changes were more obvious in group E2. Conclusions: Strenuous endurance exercise can improve level of myocardial SOD2 expression at mRNA and protein, and enhance the activity for SOD2, thus increasing the activity for T-SOD. Effect of long-term strenuous endurance exercise was better than a single bout of one.展开更多
文摘目的探究lncRNA GClnc1是否通过SIRT3/FOXO3/SOD2调控胃癌细胞增殖和迁移。方法通过转染慢病毒的方法,在人胃癌细胞SGC7901中分别过表达和沉默GClnc1,采用Western blot和qRT-PCR检测SIRT3/FOXO3/SOD2 mRNA和蛋白水平的改变;在过表达GClnc1的同时分别沉默SIRT3/FOXO3/SOD2,采用CCK-8检测细胞增殖能力,划痕实验检测细胞迁移能力。结果过表达GClnc1可以上调SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并增强SGC7901细胞的增殖[(1.00±0.14)vs(1.79±0.21),P<0.05]和迁移能力(10.87±0.76 vs 16.53±2.25,P<0.05),而沉默GClnc1则降低SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并减少细胞增殖(1.00±0.14 vs 0.62±0.13,P<0.05)和迁移能力(10.87±0.76 vs 6.46±0.89,P<0.01)。沉默SIRT3/FOXO3/SOD2可以消除过表达GClnc1对细胞增殖和迁移能力的促进作用。结论 GClnc1通过激活SIRT3/FOXO3/SOD2信号通路促进SGC7901细胞增殖和迁移。
文摘目的观察益髓灸对D-半乳糖衰老小鼠脑组织锰超氧化歧化酶(SOD2)、GADD45蛋白及其m RNA表达的影响。方法 60只SPF级雄性昆明小鼠,随机分为生理组、模型组、艾灸1组、艾灸2组和非穴位组,每组12只。采用D-半乳糖颈背部皮下注射,建立亚急性衰老小鼠模型。造模第13天艾灸1组、艾灸2组和非穴位组开始艾灸治疗,共治疗58 d。生理组及模型组不做任何治疗性干预,给予与各治疗组相同时间、程度的刺激。采用免疫组化检测海马和大脑皮质SOD2、GADD45蛋白水平,RT-PCR检测SOD2、GADD45的m RNA表达情况。结果与生理组比较,模型组海马和大脑皮质SOD2、GADD45蛋白的表达显著降低,差异具有统计学意义(P<0.05);与模型组比较,艾灸1组和艾灸2组SOD2、GADD45的蛋白表达显著升高,差异具有统计学意义(P<0.05)。与生理组比较,模型组海马和大脑皮质SOD2、GADD45 m RNA的表达显著降低,差异具有统计学意义(P<0.05);与模型组比较,艾灸1组和艾灸2组SOD2、GADD45 m RNA的表达显著升高,差异具有统计学意义(P<0.05)。结论益髓灸能够提高D-半乳糖衰老小鼠脑组织SOD2、GADD45蛋白及m RNA的表达,有提高神经细胞的抗氧化能力、修复力的作用。
文摘目的研究微小RNA(microRNA,miRNA)miR-25-3p对心肌成纤维细胞纤维化表型的调控及机制。方法血管紧张素Ⅱ(AngⅡ)灌注小鼠构建心肌纤维化模型。用miRNA表达谱芯片检测纤维化的小鼠心肌中表达水平有差异的miRNA。原代分离法获得C57BL/6小鼠心肌成纤维细胞(mCF),建立AngⅡ处理mCF的心肌纤维化细胞模型。mCF转染miR-25-3p mimic后检测纤维化相关基因的表达。双荧光素酶报告基因实验验证miR-25-3p与B细胞易位基因2(BTG2)的3′端非翻译区(3′UTR)的结合作用。放线菌素D实验验证BTG2对超氧化物歧化酶2(SOD2)稳定性的作用。结果miR-25-3p在AngⅡ灌注诱导的小鼠心肌和AngⅡ处理的mCF中表达升高。转染miR-25-3p mimic可使mCF中的Ⅰ型胶原α1(COL1A1)、Ⅲ型胶原α1(COL3A1)和α-平滑肌肌动蛋白(α-SMA)基因表达显著增加。双荧光素酶报告基因实验和功能实验证实BTG2是miR-25-3p的下游靶基因,并参与介导miR-25-3p促进mCF中纤维化相关基因表达的作用。机制研究证实BTG2可通过增加SOD2 m RNA的稳定性而上调其表达。结论miR-25-3p通过抑制BTG2的表达来下调SOD2的水平进而增强纤维化相关基因的表达,发挥促进心肌纤维化的作用。
文摘Objective: To explore the effects of myocardial SOD2 by strenuous endurance exercise. Methods: 27 grown male SD rats were randomly divided into control group (C), a single bout of strenuous endurance exercise group (E1) and seventh-week strenuous endurance exercise group (E2). Real-time PCR was used to observe the changes of mRNA expression for myocardial SOD2. Western bolt was used to observe the changes of SOD2 protein expression. In addition, SOD2, T-SOD and SOD1 activity changes were observed. Results: Myocardial SOD2 expression level at mRNA and protein of Group E1, E2 was significantly higher than that in group C, and SOD2 and T-SOD activity in group E2 were significantly higher than those in group C. Those changes were more obvious in group E2. Conclusions: Strenuous endurance exercise can improve level of myocardial SOD2 expression at mRNA and protein, and enhance the activity for SOD2, thus increasing the activity for T-SOD. Effect of long-term strenuous endurance exercise was better than a single bout of one.
