The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed...The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed in E. coil in a soluble state. The （His）6 fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass（Mr） was 88000. The results of the sequence analysis indicate that the cloned gene（GeneBank accession No. EU124675） was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid. The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides, and further to screen new herbicides.展开更多
目的研究化痰祛湿活血方对非酒精性脂肪性肝炎(NASH)大鼠脂联素(ADPN)/腺苷酸活化蛋白激酶(AMPK)/乙酰辅酶A羧化酶(ACC)通路的影响。方法 72只SD大鼠随机分为空白组、模型组、对照组及高、中、低剂量组,除空白组外,其余5组以高脂饲料联...目的研究化痰祛湿活血方对非酒精性脂肪性肝炎(NASH)大鼠脂联素(ADPN)/腺苷酸活化蛋白激酶(AMPK)/乙酰辅酶A羧化酶(ACC)通路的影响。方法 72只SD大鼠随机分为空白组、模型组、对照组及高、中、低剂量组,除空白组外,其余5组以高脂饲料联合四环素腹腔注射建立NASH模型,造模第2周分别给予化痰祛湿活血方高、中、低剂量和多烯磷脂酰胆碱(易善复)胶囊对照治疗。实验结束时,观察肝匀浆游离脂肪酸(FFA)含量,ELISA及RT-q PCR法测肝组织ADPN、脂联素受体2(AdipoR2)、AMPK、ACC、肉毒碱棕榈酰转移酶1(CPT-1)的基因与蛋白表达。多组间数据比较采用方差分析,方差齐者进一步两组间比较采用LSD-t检验,方差不齐者采用Dunnett's T3法。结果与对照组比较,高剂量组能明显降低肝匀浆FFA[(229.07±46.62)μmol/gprot vs(382.75±30.93)μmol/gprot]、肝组织ACC的基因(1.66±0.04 vs 2.40±0.37)、蛋白[(512.20±60.70)pg/g vs(756.72±61.21)pg/g]表达(P值均<0.01),升高ADPN、AdipoR2、AMPK、CPT-1的基因[1.53±0.31 vs 0.75±0.11,1.38±0.33 vs 0.61±0.09,1.43±0.39 vs 0.78±0.04,1.39±0.31 vs 0.52±0.04]、蛋白[(22.23±2.75)μg/g vs(14.01±2.17)μg/g,(128.41±10.46)ng/g vs(94.62±6.88)ng/g,(1353.79±59.52)μg/g vs(1107.19±56.78)μg/g,(175.54±9.91)U/g vs(128.74±12.74)U/g]表达(P值均<0.01)。结论化痰祛湿活血方能够有效激活ADPN/AMPK/ACC信号通路,影响脂肪酸的β氧化及合成,减少肝细胞脂肪沉积。展开更多
基金Supported by the National Natural Science Foundation of China(Nos. 20432010, 20672045 and 30570405)
文摘The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed in E. coil in a soluble state. The （His）6 fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass（Mr） was 88000. The results of the sequence analysis indicate that the cloned gene（GeneBank accession No. EU124675） was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid. The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides, and further to screen new herbicides.
文摘目的研究化痰祛湿活血方对非酒精性脂肪性肝炎(NASH)大鼠脂联素(ADPN)/腺苷酸活化蛋白激酶(AMPK)/乙酰辅酶A羧化酶(ACC)通路的影响。方法 72只SD大鼠随机分为空白组、模型组、对照组及高、中、低剂量组,除空白组外,其余5组以高脂饲料联合四环素腹腔注射建立NASH模型,造模第2周分别给予化痰祛湿活血方高、中、低剂量和多烯磷脂酰胆碱(易善复)胶囊对照治疗。实验结束时,观察肝匀浆游离脂肪酸(FFA)含量,ELISA及RT-q PCR法测肝组织ADPN、脂联素受体2(AdipoR2)、AMPK、ACC、肉毒碱棕榈酰转移酶1(CPT-1)的基因与蛋白表达。多组间数据比较采用方差分析,方差齐者进一步两组间比较采用LSD-t检验,方差不齐者采用Dunnett's T3法。结果与对照组比较,高剂量组能明显降低肝匀浆FFA[(229.07±46.62)μmol/gprot vs(382.75±30.93)μmol/gprot]、肝组织ACC的基因(1.66±0.04 vs 2.40±0.37)、蛋白[(512.20±60.70)pg/g vs(756.72±61.21)pg/g]表达(P值均<0.01),升高ADPN、AdipoR2、AMPK、CPT-1的基因[1.53±0.31 vs 0.75±0.11,1.38±0.33 vs 0.61±0.09,1.43±0.39 vs 0.78±0.04,1.39±0.31 vs 0.52±0.04]、蛋白[(22.23±2.75)μg/g vs(14.01±2.17)μg/g,(128.41±10.46)ng/g vs(94.62±6.88)ng/g,(1353.79±59.52)μg/g vs(1107.19±56.78)μg/g,(175.54±9.91)U/g vs(128.74±12.74)U/g]表达(P值均<0.01)。结论化痰祛湿活血方能够有效激活ADPN/AMPK/ACC信号通路,影响脂肪酸的β氧化及合成,减少肝细胞脂肪沉积。
