Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells     

QIN Si  ZHANG Qian  WANG Jing-jie  YU Min  LUO Yan-bei  DING Jing  LU De-qin 《园艺学报》2020,36(5):803-809

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.  相似文献   

Effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE^-/- mice     

WU Yao  SONG Nan  JIA Lian-qun  WANG Jie  CHEN Si  CAO Hui-min 《园艺学报》2020,36(7):1261-1268

AIM To observe the effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE^-/- mice. METHODS Thirty-two ApoE^-/- mice were randomly divided into model group, high-dose (60 mg/kg) tanshinone ⅡA group, low-dose (30 mg/kg) tanshinone ⅡA group and simvastatin group, and C57BL/6J mice (n=8) were used as normal control group. The mice in normal control group were given the basic feeding, while the others were given high-fat diet. The mice in tanshinone ⅡA groups and simvastatin group were given corresponding drugs. The mice in normal control group and model group were intraperitoneally injected with equal volume of saline. Eight weeks later, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were tested by automatic biochemistry analyzer. The liver tissues were stained with HE and oil red O. The contents of reactive oxygen species (ROS) and glutathione (GSH) in liver tissues of the mice were measured by commercially available kits. The liver glutathione peroxidase 4 (GPX4) and p53 were detected by immunohistochemical method. The protein and mRNA expression levels of ferroptosis-related factors GPX4, xCT/SLC7A11, p53 and ferritin heavy chain 1 (FTH1) were determined by Wes automatic Western blot quantitative analysis system and RT-qPCR. RESULTS Compared with normal control group, the serum levels of TC, TG and LDL-C in model group were increased significantly (P<0.05 or P<0.01), and HDL-C did not change significantly. The fat vacuoles were clearly visible in liver tissue. The content of ROS in liver tissue was increased significantly,and GSH was decreased significantly (P<0.01). The mRNA and protein expression levels of p53 were increased significantly, and GPX4, xCT/SLC7A11 and FTH1 were decreased significantly (P<0.05 or P<0.01). Compared with model group, tanshinone ⅡA significantly decreased the serum levels of TC, TG and LDL-C (P<0.05 or P<0.01), and HDL-C did not change significantly. High-dose and low-dose tanshinoneⅡA also significantly decreased the degree of steatosis, and the size of lipid droplets. The content of ROS in liver tissues was decreased significantly, and GSH was increased significantly (P<0.01). The mRNA and protein expression levels of GPX4, xCT/SLC7A11 and FTH1 were increased significantly, and p53 were decreased significantly (P<0.05 or P<0.01). CONCLUSION Tanshinone ⅡA reduces liver lipid deposition and lipid peroxidation damage in ApoE^-/- mice, which may be related to the intervention of ferroptosis-related proteins in the liver cells.  相似文献   

Effects of different active constituents of Gynostemma pentaphyllum on mitochondrial energy metabolism-related proteins in endothelial cells induced by ox-LDL     

ZHENG Hao-long  SONG Nan  CAO Hui-min  CHEN Si  WANG Jie  Lü Xiao-ming  JIA Lian-qun 《园艺学报》2020,36(4):713-718

AIM To investigate the effects of different components of Gynostemma pentaphyllum ［gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)］ on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a （Cox5a）, NADH:ubiquinone oxidoreductase core subunit S1 （Ndufs1）, ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C）, were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P<0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P<0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P<0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P<0.05 or P<0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis.  相似文献   

湿加松一年生母树萌条扦插效果分析   总被引：4，自引：2，他引：2  

张应中  赵奋成  李福明  钟岁英  司徒荣贵  黄永达  刘宏杰 《广东林业科技》2006,22(2):1-5

选用1年生湿加松母树萌条,分别设基部萌条与顶部促萌条、抽梢与未抽梢的顶部促萌条、顶端次生叶及穗条长度不同的顶部促萌条进行扦插试验。结果表明:(1)基部萌条与顶部促萌条、抽梢与未抽梢的顶部促萌条在扦插生根率、扦插苗根系生长、抽梢生长的差异均不显著;四类穗条扦插均可获得较高生根率,根系和抽梢生长快。(2)不同顶端次生叶长度的顶部促萌条在生根率、不定根数量上均存在极显著差异,而根系级数、须根数量及抽梢长度方面差异不显著;顶端次生叶长度0~3 cm的顶部促萌条扦插效果好。(3)不同长度的顶部促萌条制成的穗条其扦插苗抽梢长度存在极显著差异,生根率存在显著差异,根系生长差异不显著;最好选长于10 cm的顶部促萌条制成长8~10 cm的插穗扦插效果好。结合生产上扦插用育苗袋的大小及扦插深度,对1年生湿加松母株,选用顶端次生叶长3 cm以下,长8~10 cm的顶部促萌条或基部萌条扦插效果较好,要用顶端次生叶长大于3 cm的顶部促萌条扦插,只能剪取萌条长10 cm以上且制成短于10 cm的穗条扦插才能达到较好的效果。  相似文献   

基于热致液晶弹性体的仿生心脏泵的设计     

杨佳楠  陈思  王芳群  荆腾  乔筱祺 《排灌机械工程学报》2020,38(12):1221-1225

为设计一款仿生心脏功能较佳的心室泵,采用柔性、热致伸缩的热致形变弹性体代替心肌,根据螺旋心室心肌带理论,选择心肌带的左心室段,并选取了Hilbert平面填充曲线作为热致导体在热致形变弹性体中的走行方式,设计出与真实心脏类似的心室泵结构.通过试验测量了所设计的心室泵的射血分数与心率,结果显示该心室泵的搏出量高于自然心脏,而其心率远低于自然心脏.相对于机械泵,所设计的心室泵结构与真实心脏类似,具有收缩能力强,射血分数高的优点,且不易破坏血细胞,生物相容性较好.不足之处在于,热致动液晶弹性体变形响应慢,暂时还未能达到正常心脏的心率.论证了构建双稳态结构,提高仿生心室泵效率的可能性,以及对于心肌纤维走向较为复杂的其他心肌带段,双轴拉伸机械雕刻对其仿生的可能性.此研究可为容积心脏泵的研发提供新的思路,也可以对右心衰竭等心脏疾病起到辅助治疗的作用.  相似文献   

重庆地区草地贪夜蛾幼虫肠道真菌的分离鉴定          下载免费PDF全文

吴燕燕  耿敬可  徐思  唐运林  顾偌铖  韦俊宏  潘国庆  周泽扬 《西南大学学报(自然科学版)》2019,41(9):1-8

草地贪夜蛾(Spodoptera frugiperda)是一种洲际迁飞性农业害虫,食谱广泛. 2019年5月已迁飞至重庆地区,对农作物造成严重危害.为了解入侵重庆地区草地贪夜蛾(Spodoptera frugiperda)肠道真菌菌群的组成,本研究以采集自重庆巫山玉米田及重庆江津高粱田中的草地贪夜蛾幼虫为材料,运用传统培养方法分离其粪便中的肠道优势真菌,并对其进行形态学观察和分子生物学鉴定.实验通过分离培养不同地区、不同食物的草地贪夜蛾肠道真菌,结合形态学观察及rDNA ITS测序完成了属水平的鉴定,共分离得到了5个属10个真菌分离株,其中分离自重庆巫山玉米田的草地贪夜蛾肠道真菌归为3个属,分别为念珠菌属(Candida)、掷孢酵母属(Sporobolomyces)和帚枝霉属(Sarocladium);分离自重庆江津高粱田的草地贪夜蛾肠道真菌归为3个属,分别为念珠菌属(Candida)、莫氏黑粉菌属(Moesziomyces)和毛霉属(Mucor).初步分离获得入侵重庆地区的草地贪夜蛾肠道真菌,丰富了对入侵草地贪夜蛾肠道微生物的认识.  相似文献   

不同稳定剂对胭脂萝卜红色素稳定性的影响     

芮春梅  李成琼  司军  宋洪元  任雪松  李利军 《勤云标准版测试》2006,(12)

天然胭脂萝卜红色素属于花青素类化合物,性质不稳定。笔者以新鲜胭脂萝卜(Raphnussativuscv.)肉质根为原料,提取胭脂萝卜红色素,选择常用的5种稳定剂对其稳定性进行研究,从而确定出最佳的稳定剂及其用量。研究结果发现0.03%维生素C、0.03%谷氨酸、0.03%柠檬酸能明显地延缓其颜色变化,0.03%维生素C 0.03%柠檬酸的组合对该色素的稳定效果最佳;进一步研究确定维生素C和柠檬酸的最佳用量分别为0.04%和0.08%。  相似文献   

SDS-PAGE和PCR检测1B/1R易位系研究初报     

司红起  马传喜 《勤云标准版测试》2006,(7)

采用18个冬小麦品种作为供试材料,分别通过SDS-PAGE检测黑麦Secalin蛋白和PCR检测黑麦1RS染色体或其片段,结果表明:SDS-PAGE检测不出不带有secalin蛋白的1B/1R小片段易位系,而PCR则能检测出,从而提高1B/1R易位系的检出率;SDS-PAGE和PCR结合是检测1B/1R易位系的一种快速经济的有效方法。  相似文献   

甘肃省草地资源退化原因及草地生态系统恢复途径     

王月梅  张金林  司宗信 《勤云标准版测试》2006,(8)

提出甘肃省草地资源退化状况,分析得出人为破坏、草地超负荷利用、草地鼠虫害频繁发生、草地群落物种数减少、草地生态系统生产力下降、不合理的开矿、草地保护、管理意识不强等因素导致了甘肃省草地资源的日益退化。最后指出要恢复草地生态系统,必须从以下七个途径入手,既①强化宣传教育,依法管护草地资源;②科学规划,分类指导;③规模种植,加快人工草地建设;④加强围栏封育;⑤大力实施草地改良;⑥依靠科技进步,保护和建设草地资源;⑦从空中要水。这将对甘肃省乃至西北地区草地生态系统的恢复与重建提供一定的参考。  相似文献   

H6N2亚型禽流感病毒血凝素基因的克隆及其序列分析     

陈思怀  乔宪凤  华文君  郑新民  魏庆信  黎晓敏  张涌 《西南大学学报(自然科学版)》2006,28(5):846-850

设计特异引物,采用RT—PCR技术,成功从本实验室分离、保存的H6N禽流感病毒毒株（HB2002）中克隆到血凝素基因（HA）序列,该克隆基因全长1714hp,提交GenBank,获得登录号DQ285546。同部分H6,H5,H7,H9亚型禽流感病毒HA基因序列分析表明,该基因核酸、氨基酸序列与A／duck／Hainan／4／2004（H6N2）,A／duck／HongKong／3600／99（H6N2）毒株有99％的相似性;分子进化分析结果显示,该基因与此2毒株亲缘关系最近。由该克隆核酸序列推导的HA蛋白共566个氨基酸,该蛋白同其他H6亚型AIV的HA蛋白有相同裂解位点序列即PQIETR↓G,无高致病性毒株典型的裂解位点特征,因此,初步判定毒株HB2002为低致病性禽流感病毒．  相似文献