Effect of hypercapnia on lysyl oxidase-dependent collagen cross-linking and hypoxia-induced pulmonary hypertension in rat

AIM: To investigate the effect of hypercapnia on hypoxia-induced pulmonary hypertension and the changes of lysyl oxidase (LOX) and extracellular matrix collagen cross-links in the rat. METHODS: Sprague-Dawley rats were randomly divided into 4 groups:normoxia group, hypoxia group, hypercapnia group and hypoxia+hypercapnia group. LOX activity was detected by fluorescence spectrophotometry. LOX protein expression was detected by immunohistochemistry and Western blot. The mRNA expression of LOX in the pulmonary artery was detected by real-time PCR. RESULTS: The levels of mean pulmonary artery pressure (mPAP), RV/(LV+S) and WA/TA in hypoxia group were significantly higher than those in normoxia group (P<0.01). Moreover, the levels of mPAP and RV/(LV+S) in hypoxia+hypercapnia group were significantly lower than those in hypoxia group (P<0.01). However, no significant difference of mPAP and RV/(LV+S) between hypercapnia group and normoxia group was observed. In hypoxia group, the collagen cross-links in the lung tissue was significantly higher than that in normoxia group and hypercapnia group (P<0.01). Importantly, collagen cross-links in the lung tissue of hypoxia+hypercapnia group was significantly lower than that in hypoxia group (P<0.01). There was no significant difference in collagen cross-links between hypercapnia group and normoxia group. The expression of LOX at mRNA and protein levels and its activity in the pulmonary arteries of hypoxia group were significantly increased as compared with normoxia group (P<0.01). Furthermore, the expression of LOX at mRNA and protein levels and its activity in the pulmonary arteries in hypoxia+hypercapnia group were lower than those in hypoxia group (P<0.01). CONCLUSION: Hypoxia not only up-regulates LOX but also promotes collagen cross-linking in the rat lung, which contributes to the development of pulmonary hypertension. Hypercapnia inhibits hypoxia-induced LOX expression and collagen cross-linking, therefore impairing the progress in hypoxia-induced pulmonary hypertension.     

根际氧浓度对番茄幼苗生长发育的影响

为了探究设施园艺作物栽培时适宜的水气环境,采用水培试验,以河南四号和粉珍珠2个栽培番茄品种为研究对象,设置3个氧水平:低氧(0.5~2.0 mg/L)、正常氧(7.0~8.0 mg/L)和饱和氧(14.0~17.0 mg/L)处理,以不通气作为空白对照CK,研究根际不同氧浓度对番茄幼苗植株的形态生长、生物量、根系活力和光合色素的影响。结果表明,随着根际氧浓度的增加,番茄幼苗的长势越好,根系活力增加,但叶绿素含量降低。在处理末期,正常氧处理与饱和氧处理显著提高番茄幼苗的最长根长并增加番茄幼苗的干、鲜质量。其中,在18 d河南四号正常氧和饱和氧处理下总干质量比CK分别增加24.00%和74.86%,而低氧处理比CK降低3.43%;最长根长分别增加67.23%和230.55%,而低氧处理比CK降低1.63%。饱和氧处理下根系更发达健壮,地上部分长势更好,生物量积累最多。因此,在设施园艺栽培管理中,增加营养液中根际氧浓度有利于番茄生物量的积累,进而提高产量。     

Chronic hypoxia enhances aggressiveness of MCF-7 breast cancer cells through EMT

AIM: To investigate the effects of chronic hypoxia on the aggressiveness of MCF-7, a human breast cancer cell line, and the underlying mechanisms.METHODS: MCF-7 cells were cultured under hypoxia (1% O₂, 5% CO₂ and 94% N₂) or control (95% O₂ and 5% CO₂) condition. The viability, proliferation, and invasion and migration abilities of the MCF-7 cells were determined by MTT assay, CCK-8 assay, cell counting, and cell invasion and migration assays. Anchorage-independent growth and the alteration of cellular polarization of the MCF-7 cells were tested by soft agar colony formation assay and Matrigel-3D culture assay, respectively. The effects of chronic hypoxia on the growth and metastasis of MCF-7 cells in vivo were investigated by xenograft in nude mice. The morphological changes of the MCF-7 cells were observed under an inverted microscope. Hypoxia-induced alterations in the levels of hypoxia inducible factor-1 (HIF-1) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β) as well as epithelial-mesenchymal transition (EMT) molecules, such as E-cadherin, N-cadherin, vimentin, matrix metalloproteinase (MMP)-3 and MMP-9, were determined by Western blot.RESULTS: Chronic hypoxia significantly increased the viability, proliferation, and invasion and migration abilities of MCF-7 cells in vitro, enhanced the anchorage-independent growth, facilitated cellular polarization alteration in Matrigel-3D culture, and promoted cancer metastasis in vivo. Hypoxia up-regulated HIF-1, activated GSK-3β, down-regulated E-cadherin and increased the protein levels of N-cadherin, vimentin, MMP-3 and MMP-9. CONCLUSION: Chronic hypoxia enhances the aggressiveness of breast cancer cells probably through EMT.     

低溶氧胁迫对刺参(Apostichopus japonicus)氧化应激指标的影响

为探明低氧胁迫对刺参(Apostichopus japonicus)抗氧化能力的影响以及刺参的低氧逆境响应机制，给低氧环境条件下的刺参养殖提供指导，本研究通过设置低氧胁迫实验，将刺参在水体低氧[(2.0±0.2) mg/L]8 h 处理后恢复常氧[(7.0±0.2) mg/L]2.5 h，取低氧和常氧不同时间段的刺参肌肉、呼吸树和消化道组织，对各组织的乳酸(LD)、丙二醛(MDA)和抗氧化酶系等应激参数进行测定和变化趋势分析。结果显示，与对照组相比，在低氧胁迫8 h 内，随着低氧暴露时间的延长，刺参肌肉、呼吸树和消化道等组织中的 LD 含量、抗氧化能力(T-AOC)、过氧化氢酶(CAT)、谷胱甘肽-S-转移酶(GST)活力显著上升；超氧化物歧化酶(SOD)活力显著下降；肌肉组织中的 MDA 含量显著降低，呼吸树和消化道中的 MDA 含量显著上升。在恢复常氧阶段，各氧化应激指标逐渐恢复到正常水平。低氧胁迫使刺参的有氧代谢减弱，无氧代谢增强，以维持机体能量需求。低氧胁迫造成刺参机体各种应激生化指标上升或下降，这是机体为适应低氧环境刺激而作出的一种抗氧化策略。     

Effect of ROCK1 and ROCK2 silencing by shRNA on hypoxia-induced apoptosis of rat cardiomyocytes

AIM: To investigate the effects of Rho-associated coiled-coil protein kinase-1 (ROCK1) and ROCK2 on apoptosis induced by hypoxia in rat cardiomyocytes. METHODS: Rat cardiomyocytes were cultured primarily and identified using an antibody targeting α-actin of striated muscle. ROCK1-shRNA and ROCK2-shRNA were transiently transfected into the cells by liposome. After 48 h, these cells were subject to hypoxia for 6 h. The cells were divided into 5 groups: blank control group, hypoxia group, hypoxia+negative control shRNA group, hypoxia+ROCK1-shRNA group and hypoxia+ROCK2-shRNA group. The beating frequency and rhythm of the cardiomyocytes were assessed by microscopy. The activity of lactate dehydrogenase (LDH) in the cell culture supernatants was detected by automatic biochemical analyzer. The cell survival rate was analyzed by the method of MTT. The cell apoptotic rate was assessed by flow cytometry. Western blotting was used to determine the expression of ROCK1, ROCK2, caspase-3 and p-PI3K. RESULTS: The primary culture of the cardiomyocytes was successful. Western blotting results showed that the transfection of ROCK1-shRNA or ROCK2-shRNA decreased the expression of ROCK1 or ROCK2 in the cardiomyocytes. Hypoxia slowed down the beat frequency of the cardiomyocytes, also made the rhythm disorder. Hypoxia increased the release of LDH and decreased the cell survival rate. Flow cytometry results showed that hypoxia increased the cell apoptotic rate. Hypoxia increased the expression of caspase-3 and decreased the expression of p-PI3K. Transfection of ROCK1-shRNA and ROCK2-shRNA into the cardiomyocytes reduced all the effects of hypoxia mentioned above. CONCLUSION: Down-regulation of ROCK1 and ROCK2 expression suppresses the apoptosis of rat cardiomyocytes induced by hypoxia. The mechanism is associated with the inhibition of caspase-3 activation and the up-regulation of p-PI3K expression.     

Modeling the impact of alternative drainage practices in the northern Corn-belt with DRAINMOD-NII

The hydrologic and water quality impacts of subsurface drainage design and management practices are being investigated through field and simulation studies throughout the northern Corn-belt. Six years of data from an ongoing field study in south central Minnesota (Sands et al., 2008) were used to support a modeling effort with DRAINMOD-NII to investigate: (1) the performance of the model in a region where soils are subject to seasonal freeze-thaw and (2) the long-term hydrologic and water quality characteristics of conventional and alternative subsurface drainage practices. Post-calibration model prediction and efficiency were deemed satisfactory using standard model performance criteria. Prediction errors were primarily associated with early spring snowmelt hydrology and were attributed to the methods used for simulating snow accumulation and melting processes, in addition to potential sublimation effects on ET estimates. Long-term simulations with DRAINMOD-NII indicated that drainage design and/or management practices proposed as alternatives to conventional design may offer opportunities to reduce nitrate (NO₃)-nitrogen losses without significantly decreasing (and in some cases, increasing) crop yields for a Webster silty clay loam soil at Waseca, Minnesota. The simulation study indicated that both shallow drainage and controlled drainage may reduce annual drainage discharge and NO₃-nitrogen losses by 20-30%, while impacting crop yields from −3% (yield decrease) to 2%, depending on lateral drain spacing. The practice of increasing drainage intensity (decreasing drain spacing) beyond recommended values appears to not significantly affect crop yield but may substantially increase drainage discharge and nitrate-nitrogen losses to surface waters.     

Effects of HIF-1α/SDF-1 signaling axis on hypoxia-induced migration and adhesion of progenitor cells

AIM: To observe the expression of hypoxia-inducible factor-1alpha (HIF-1α) and stromal cell-derived factor 1 (SDF-1) in pulmonary artery endothelial cells (PAECs) of SD rats and to investigate the role of HIF-1α/SDF-1 signaling axis on hypoxia-induced migration and adhesion of progenitor cells to PAECs. METHODS: Immunomagnetic beads were used to separate and purify the CD34⁺/CXCR4⁺ progenitor cells derived from the peripheral circulation of SD rats. The expression of HIF-1α and SDF-1 in PAECs exposed to hypoxia (1% O₂, 5% CO₂ and 94% N₂) was detected by immunofluorescence, Western blotting and ELISA. The migration index and adhesion rate were measured in the progenitor cells, which were subjected to the following different treatments: (1) normoxia (21% O₂); (2) hypoxia 12 h; (3) hypoxia 12 h +HIF-1α inhibitor (2ME2); (4) hypoxia 12 h+SDF-1 neutralizing antibody; (5) hypoxia 12 h+2ME2+SDF-1 neutralizing antibody.RESULTS: The expression of HIF-1α and SDF-1 in PAECs was effectively induced by the hypoxic exposure, and both of them reached the peak levels after 12 h of hypoxic treatment (P<0.01), while administration of 2ME2 decreased the hypoxia-induced SDF-1 expression (P<0.05). Treatment of the PAECs with 2ME2 or SDF-1 neutralizing antibody attenuated the migration index and adhesion rate of progenitor cells to the PAECs (P<0.05). CONCLUSION: There is a HIF-1α/SDF-1 signaling axis in hypoxia-exposed PAECs, which may play a crucial role in the migration and adhesion of progenitor cells to PAECs.     

耗氧率及溶氧胁迫对长蛸体内酶活力的影响

采用Winkler法测定水中溶氧含量,通过比较对照呼吸室与试验呼吸室水中溶解氧含量之差,测定长蛸(Octopus variabilisSasaki)耗氧率及窒息点,并在不同程度的溶氧胁迫下测定长蛸体内多种酶的活力变化。结果表明,长蛸耗氧率在一个昼夜内呈现低-高-低-高的趋势;耗氧率与水温(10~30℃)呈正相关,与pH值(6.40~9.20)呈负相关;随着盐度的升高(14~31),长蛸耗氧率表现为两头高、中间平稳,曲线呈凹状分布;雌性长蛸耗氧率高于雄性。窒息点随长蛸体重的增加而降低,平均体重为(30.50±6.03)g的长蛸窒息点为0.4179mg/L;平均体重为(54.50±4.65)g的长蛸窒息点为0.3902mg/L;平均体重为(97.00±11.36)g的长蛸窒息点为0.2738mg/L。随着溶氧胁迫程度的增加,LDH活力呈先升后降趋势,脂肪酶活力下降,SOD、POD、CAT、ACP、AKP和蛋白酶活力均呈降-升-降的趋势。     

24-表油菜素内酯对低氧胁迫下黄瓜幼苗根系抗氧化系统的影响

 【目的】探讨外源EBR（24-表油菜素内酯）对低氧胁迫下黄瓜（Cucumis sativus L.）根系的生长和抗氧化系统的影响。【方法】采用营养液水培法，以抗低氧能力较强的黄瓜品种绿霸春4号和抗低氧能力较弱的品种中农8号为材料，研究根际低氧胁迫下外源施用EBR对黄瓜根系生长、ROS及抗氧化酶活性的影响。【结果】在低氧胁迫下，EBR处理显著促进了低氧胁迫下两品种黄瓜幼苗根系生长，提高了SOD、APX、GR活性及AsA、GSH含量，降低了O 、H2O2及MDA含量。【结论】外源EBR处理通过促进低氧胁迫下黄瓜幼苗根系中抗氧化酶活性和抗氧化剂含量的提高，降低ROS含量，增强植株抗低氧胁迫的能力。     

Effects of different β-adrenergic receptors on cardiac systolic and diastolic functions in hypoxic stress rats

AIM:To explore the effects of different β-adrenergic receptors (β-AR) on the left and right ventricular systolic and diastolic functions in rats under acute hypoxic stress. METHODS:The healthy male SD rats were randomly divided into 4 groups (n=7):control group, non-selected β-AR blocker propranolol group, selected β₁-AR blocker atenolol group and selected β₂-AR blocker ICI 118,551 group, and then the rats were exposed to normoxia (20.9% O₂, 79.1% N₂) and hypoxia (15.0% O₂, 85.0% N₂) condition respectively at the altitude of 2 260 m (Xining, China). The heart rate (HR), the left ventricular systolic pressure (LVSP), the right ventricular systolic pressure (RVSP), and the maximum raise/decline rate of left and right ventricular pressure (±dp/dt_max) were monitored, and the arterial blood gas in normoxia and hypoxia condition were compared to explore the effect of β-AR on the left and right ventricular systolic and diastolic functions in acute hypoxic stress rats. RESULTS:Under normoxia condition, the LVSP, ±dp/dt_max of left ventricular were decreased in propranolol group, atenolol group and ICI 118,551 group, the RVSP and ±dp/dt_max of right ventricle were decreased in propranolol group and atenolol group (P<0.05). Under hypoxia condition, the PaO₂, LVSP, ±dp/dt_max of left ventricle were decreased in all groups compared with the normoxia group, and the ±dp/dt_max of right ventricle was increased in all groups (P<0.05), also the degree of index change in control group was more obvious than that in propranolol group and atenolol group. CONCLUSION:The activation of β₁-AR is an important compensatory regulation for heart function during hypoxic stress. However, the compensatory enhancement of right heart function under acute hypoxia condition which through tonogenic dilation is more significant for maintaining the normal circulating blood flow.