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

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.     

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

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.     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

不同基质配比对温室番茄幼苗生长及叶绿素荧光参数的影响

以番茄品种"福美十号"为材料,采用随机区组试验设计,研究不同基质配比对番茄幼苗生长与叶片叶绿素荧光参数的影响。结果表明:采用A3[V(草炭)∶V(玉米秸秆)∶V(牛粪)∶V(食用菌下脚料)∶V(蛭石)=1∶2∶0.5∶0.5∶1]的基质配比,幼苗在出苗率、株高、茎粗、叶片数和根冠比等性状上表现最优,分别为98.25%、44.60 cm、0.50 cm、9.12片与7.95;叶片光系统Ⅱ(PSIⅡ)原初光能转化效率(Fv/Fm)与PSⅡ潜在活性(Fv/Fo)的值在幼苗生长前期、中期和末期也均为最高,Fv/Fm值分别为0.82、0.74和0.79,Fv/Fo值分别为3.92、2.97和3.23;其叶片内丙二醛(MDA)含量最低为6.02μmol/g,其根系活力最强、超氧化物歧化酶(SOD)、过氧化物酶(POD)与可溶性蛋白含量等指标最高,分别为158.26μg/(g·h)、386.23 U/g、6.26 U/(g·min)和0.203 mg/g,显著高于其他处理。说明采用处理A3的基质配比更适宜于番茄幼苗的生长。     

芝麻棒孢叶斑病病原菌鉴定与致病力比较

 为明确芝麻叶斑病病原菌及其致病力,本研究以河南不同地区芝麻上分离所得叶斑病分离物为研究对象,通过形态观察和分子鉴定探讨这些分离物的分类属性,并且进行离体叶片和活体茎杆接菌,进一步探究了不同菌株的致病力差异。结果显示分离所得的6株芝麻叶斑病分离物的菌落形态、生长速率和产孢量存在差异,分生孢子多数呈倒棍棒状,具隔膜,6个菌株的ITS序列与GenBank中Corynespora cassiicola的一致性达99%以上,结合形态特征与ITS序列将其鉴定为山扁豆生棒孢。室内人工接菌芝麻发现,这6个菌株的致病力存在显著的差异;在芝麻叶片上,20180909-03属强致病力菌株,20180821-01、20180909-05和20180824-01属中等致病力菌株,20180904-02-02和SF1-1属弱致病力菌株;在芝麻茎杆上,20180824-01致病力最强,其次是20180821-01、20180909-03和20180909-05,SF1-1和20180904-02-02致病力最弱。研究结果证明山扁豆生棒孢不同菌株在芝麻上的致病力存在明显的分化。     

我国云南、湖北和山东葡萄产区霜霉病菌对甲霜灵的抗性检测

为明确我国云南省宾川县、湖北省公安县和山东省烟台市葡萄产区霜霉病菌对甲霜灵的抗性发生态势,采用叶盘漂浮法测定了这3个产区共127株葡萄霜霉病菌对甲霜灵的抗性频率及抗性水平。结果显示,不同区域间病菌的抗性频率和抗性水平均存在差异。其中,湖北省公安县霜霉病菌的抗性频率和抗性水平均较高,抗性频率达92.0%,高抗菌株占76.0%,低抗菌株占16.0%,敏感菌株占8.0%;山东省烟台市霜霉病菌的抗性频率为74.0%,低抗菌株占64.0%,高抗菌株占10.0%,敏感菌株占26.0%;云南省宾川县霜霉病菌的抗性频率和抗性水平均较低,抗性频率为29.6%,敏感菌株占70.4%,低抗菌株占29.6%,无高抗菌株。     

养殖场粪污堆存中3种氨挥发测定技术比较研究

养殖场粪污堆放存储中的氨挥发损失是粪肥N素养分损失的重要环节。采取控氨措施能够降低粪污堆存过程中的氨排放、保存粪肥养分。从现有的可用于测定粪污堆存过程氨挥发的技术中筛选出简单易行且可靠的技术手段可为我国养殖场氨排放测定技术标准的完善提供重要依据。本研究采用通气法(VCT)、间歇式密闭室抽气法(DCT)和被动采样法(PST)同步测定牛粪堆存过程中未经处理、表面覆盖(醋糟、CZ)及施用外源添加剂(磷酸二氢钾+氯化镁、TJ)的氨挥发特征及排放通量。结果表明:(1)试验期间VCT、PST测定的氨挥发总量均低于DCT测定结果。VCT法下CK、CZ和TJ处理的氨挥发量仅为DCT法测定值的20.46%、10.09%和11.17%,而PST法下CK、CZ和TJ处理的氨挥发量为DCT法测定值的64.38%、68.64%和61.67%;(2)采用VCT、DCT、PST 3种方法测定两类减排措施的减排效果一致,覆盖醋糟(CZ)及施用外源添加剂(TJ)均可起到抑制氨挥发的作用,CZ的减排率为35.83%～70.33%,TJ的减排率为35.89%～65.01%;(3)利用PST法测定所得的氨排放量及减排率与基准方法 DCT测定结果基本一致,表明PST法可作为检验农业源氨减排措施效果的测定技术,但仍需进行更多的田间验证,为后续推广提供数据支撑。     

基于生态足迹成分法的太子山国家森林公园旅游承载力研究

运用生态足迹成分法,基于太子山国家森林公园旅游统计数据和实地调查数据,计算了该森林公园的旅游生态足迹、旅游生态承载力以及旅游可持续发展度。结果显示:2018年太子山森林公园总旅游生态足迹为1 013.36 hm~2,总旅游生态承载力为10 738.99 hm~2,总旅游生态盈余为9 725.63 hm~2,旅游可持续发展度为0.09,表明该森林公园处于生态盈余和可持续发展的理想状态。     

高炭基肥配施氮肥对牡丹江烤烟生长及产量和质量的影响

采用田间试验,设置4个处理[CK(常规施氮量,不施高炭基肥)、T1(常规施氮量,900 kg·hm~(-2)高炭基肥)、T2(85%常规施氮量,900 kg·hm~(-2)高炭基肥)、T3(70%常规施氮量,900 kg·hm~(-2)高炭基肥)],研究了减氮配施高炭基肥对牡丹江烤烟生长和产量、质量的影响。结果表明,T1处理烤烟各生育时期的农艺性状均有所提高,T3处理对烤烟的生长产生了负面效应;T1处理显著提高了中上部叶总氮含量(1.59%,1.65%)、烟碱含量(1.71%,1.99%),T1和T2处理显著提高了中部叶钾含量(1.68%,1.69%);与CK相比,T1处理中上部叶感官评吸总得分均有显著提高,增幅分别为4.06%和5.92%,其烟叶产量、产值及中上等烟比例均有所提高。在牡丹江烟区白浆土条件下,控制总施氮量为45 kg·hm~(-2)、高炭基肥料施用量为900 kg·hm~(-2)时,有利于促进烤烟生长、提高经济效益、改善烟叶品质。