沼液兼养培养微藻潜力及不同有机碳源影响

利用沼液培养微藻可在收获藻生物质的同时回收碳、氮、磷养分,是沼液资源化利用极具潜力的途径。与光合自养相比,兼养培养可实现藻生物量快速积累,且对光、碳利用灵活,与透光性不佳的沼液相性较好,但目前缺乏相关研究论证其可行性。本文首先选取了小球藻 Chlorella sp.、蛋白核小球藻 Chlorella pyrenoidosa、栅藻 Scenedesmus sp.,以葡萄糖为碳源利用猪粪沼液对 3株微藻进行了兼养培养。结果显示,兼养策略可在大幅强化藻生物量积累的同时协同提升沼液污染物去除。其中,Chlorella pyrenoidosa展现出最佳的生物量及养分去除优势,培养7 d生物量可达1.51 g·L^-1,为光合自养的6.12倍,沼液COD、氨氮、总氮、总磷去除率较自养分别提高了20、36、41个和32个百分点。本研究进一步考察了具备两种典型碳代谢路径的有机碳源（葡萄糖-三羧酸循环,乙酸钠-乙醛酸循环）对Chlorella pyrenoidosa沼液兼养培养的影响,发现葡萄糖相较于乙酸钠更适宜作为沼液兼养培养的有机碳源,且葡萄糖浓度与利用效率呈负相关,1 g·L^-1葡萄糖浓度条件下Chlorella pyrenoidosa具有最高的单位有机碳生物量产率。此外,兼养微藻通过代谢葡萄糖可协同提升光合性能,使PSⅡ最大量子产量、实际量子产量、调节性能量耗散量子产量等维持较高水平,既弥补了沼液弱透光下光能不足,也强化了持续光照后的光系统损伤恢复机制。因此,本研究认为以添加1 g·L^-1葡萄糖的沼液兼养培养Chlorella pyrenoidosa是克服沼液养藻光衰减等不利因素,强化微藻生物量产量及养分回收效率的有效方式,在畜禽养殖场沼液生物消纳与资源化利用方面有较好的应用前景。     

乙酸钠对油酸诱导的BRL-3A细胞脂肪变性的影响及机制分析

通过向油酸诱导的大鼠肝成纤维细胞(BRL-3A)脂肪变性模型中添加不同浓度(2、4、8 mmol·L^-1)的乙酸钠,探讨其对脂肪变性细胞模型脂代谢的调控机理及细胞损伤的修复作用。试验方法：1)用不同浓度的油酸(0、0.03、0.06、0.12、0.24、0.48) mmol·L^-1刺激BRL-3A细胞24 h后,分别检测细胞相对活力、总脂滴面积、三酰甘油(TG)含量、天门冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)活性,建立脂肪变性细胞模型;2)向BRL-3A细胞中添加不同浓度的乙酸钠,通过流式细胞术检测细胞凋亡率;3)用不同浓度的乙酸钠和0.12 mmol·L^-1油酸共同孵育BRL-3A细胞,试验分为4组,分别为油酸处理组、2 mmol·L^-1乙酸钠+油酸处理组、4 mmol·L^-1乙酸钠+油酸处理组和8 mmol·L^-1乙酸钠+油酸处理组,分别对细胞脂滴、TG含量、AST、ALT活性、AMPK信号通路蛋白以及脂代谢关键基因进行检测。结果显示:1)用0.12 mmol·L^-1油酸处理BRL-3A细胞24 h,成功建立BRL-3A细胞脂肪变性模型。2)不同浓度的乙酸钠对BRL-3A细胞凋亡率没有影响;3)4、8 mmol·L^-1乙酸钠处理脂肪变性细胞模型后,与油酸处理组相比,细胞总脂滴面积、每平方毫米脂滴数、TG含量、AST和ALT活性均显著(P<0.05)或极显著(P<0.01)下降,P-AMPK表达水平显著(P<0.05)或极显著(P<0.01)上升;脂合成代谢相关基因ACC、FAS以及SCD-1 mRNA表达水平均有一定程度下降;脂分解代谢相关基因CPT-1、CPT-2以及ACO mRNA表达水平均有一定程度上升。本研究表明,乙酸钠会通过AMPK通路激活脂分解代谢,减轻肝细胞脂质蓄积,并且对油酸诱导的BRL-3A细胞脂肪变性模型的损伤具有一定缓解作用。     

瘤胃乙酸、丙酸比例对绵羊体内氧化代谢和血浆胰岛素水平动态变化的影响

本研究以全胃营养灌注绵羊为试验动物 ,用物质代谢区室分析方法和同位素示踪技术研究了瘤胃乙酸、丙酸比例对体内氧化代谢和血浆胰岛素水平动态变化的影响。不同试验处理间营养灌注液能量、蛋白质水平相同 ,而瘤胃灌注混合挥发性脂肪酸的乙酸、丙酸比例不同。三种混合挥发性脂肪酸的乙酸、丙酸比例分别为75 :15(VFA1)、65 :25(VFA2)、45 :45(VFA3)。由测定引入 14C -乙酸后血液二氧化碳放射比强度—时间曲线上升段斜率(k值)反映体内氧化代谢强度。试验结果为 ,瘤胃灌注VFA1 时的k值显著高于灌注VFA2 和VFA3 时的k值(P<0.05) ,而VFA2 和VFA3 两处理间的差异不显著(P>0.05) ,说明瘤胃灌注VFA1 时绵羊体内氧化代谢强度增加。向停止灌注24小时的绵羊瘤胃内灌注VFA1,血浆胰岛素水平在恢复灌注后缓慢上升。将瘤胃灌注混合挥发性脂肪酸由VFA1 换为VFA2 后 ,血浆胰岛素水平迅速上升 ,之后迅速下降 ,而由VFA2 换为VFA3 后未出现血浆胰岛素水平的明显变化 ,说明血浆胰岛素水平的变化与体内糖代谢调节状态有关 ,而瘤胃灌注VFA1 时的体内糖代谢调节状态与灌注VFA2 和VFA3 时明显不同。     

Evidence for functional G-coupled protein receptors 43 and 120 in subcutaneous and intramuscular adipose tissue of Angus crossbred steers

We conducted 3 independent experiments to demonstrate functional G-coupled protein receptor 43 (GPR43) and GPR120 in bovine intramuscular (i.m.) and subcutaneous (s.c.) adipose tissues. We hypothesized that media volatile fatty acids and long-chain fatty acids would affect cAMP-activated protein kinase-alpha (AMPKα) protein expression and cAMP concentrations differently in i.m. and s.c. adipose tissue. Experiment 1: oleic acid (18:1n-9) decreased phosphorylated AMPKα protein (p-AMPKα) and the p-AMPKα/AMPKα protein ratio in i.m. preadipocytes, increased the p-AMPKα/AMPKα protein ratio in bovine satellite cells, and had no effect in s.c. preadipocytes. Experment 2: ex vivo explants from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers were cultured 48 hr in media containing 0.25 µM ciglitizone, 5 mM glucose, and 5 mM acetate, in the absence or the presence of 100 µM oleic acid. Oleic acid increased acetate incorporation into fatty acids and GPR43 gene expression in i.m. adipose tissue (P < 0.05), but oleic acid had no effect on fatty acid synthesis or GPR43 expression in s.c. adipose tissue. Experiment 3: fresh s.c. and i.m. adipose tissue from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers was transferred immediately to 6-well culture plates containing 3 mL of KHB/Hepes/5 mM glucose. Samples were preincubated with 0.5 mM theophylline plus 10 μM forskolin for 30 min, after which increasing concentrations of acetate or propionate (0, 10⁻³, 10^−2.3, and 10⁻³ M) in the absence or the presence of 100 μM oleic acid or 100 µM palmitic acid (16:0) were added to the incubation media. Acetate had no effect on forskolin-stimulated cAMP production in s.c. adipose tissue but decreased cAMP in i.m. adipose tissue (P < 0.05); this indicates a functional GPR43 receptor in i.m. adipose tissue. The combination of 10⁻² M acetate and oleic acid decrease cAMP production in s.c. adipose tissue, consistent with GPR120 receptor activity, but oleic acid and palmitic acid attenuated the depression of cAMP production caused by acetate in i.m. adipose tissue. Palmitic acid depressed cAMP production in s.c. adipose tissue, and increased cAMP production in i.m. adipose tissue (P < 0.05). Propionate had no effect on cAMP production in s.c. or i.m. adipose tissue. These results provide evidence for functional GPR43 receptors in i.m. adipose tissue and GPR120 receptors in s.c. adipose tissue, both of which would suppress lipolysis.     

乙酸镁对氟烷基因杂合子育肥猪肌肉加工品质的影响

育肥猪屠宰前饲粮中短期添加乙酸镁,研究其对氟烷基因杂合子育肥猪肌肉加工品质的影响。选用体重相近的氟烷基因杂合子(HalNHaln)大长猪(大白猪×长白猪)36头随机分为2组,即对照组、乙酸镁组,每组18头。乙酸镁组在屠宰前5 d,基础日粮中添加镁1 500mg/kg(以乙酸镁形式),对照组饲喂基础日粮。试验表明:在猪屠宰前饲粮中短期添加1 500 mg/kg水平的镁对试验猪的平均日采食量没有明显影响(P>0.05);可以显著降低肌肉的滴水损失(P<0.05),提高猪肉的熟肉率,从而提高猪肉的加工品质。     

铅对SD大鼠成骨细胞磷脂酰胆碱特异性磷脂酶C的影响

在体外培养新生sD乳鼠的成骨细胞中暴露不同浓度的醋酸铅（O、20、40、80μmol／L）,作用24h后检测细胞磷脂酰胆碱特异性磷脂酶C（PC-PLC）活性与PC—PLC蛋白的表达量。结果表明：不同浓度的醋酸铅均可使成骨细胞PC—PLC活性和PC-PLC蛋白表达量极显著降低（P〈0．01）,呈明显的剂量-效应关系,表明成骨细胞铅暴露可通过抑制PC—PLC而发挥毒性作用。     

Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways

Pseudorabies virus (PRV) infection leads to severe inflammatory responses and tissue damage, and many natural herbs exhibit protective effects against viral infection by modulating the inflammatory response. An ethyl acetate fraction of flavonoids from Polygonum hydropiper L. (FEA) was prepared through ethanol extraction and ethyl acetate fractional extraction. An inflammatory model was established in RAW264.7 cells with PRV infection to evaluate the anti-inflammatory activity of FEA by measuring cell viability, nitric oxide (NO) production, reactive oxygen species (ROS) release, and mRNA expression of inflammatory factors, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Its functional mechanism was investigated by analyzing the phosphorylation and nuclear translocation of key proteins in the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings indicate that PRV induced inflammatory responses in RAW264.7 cells, and the responses were similar to that in lipopolysaccharide (LPS)-stimulated cells. FEA significantly suppressed NO synthesis and down-regulated both expression and secretion of COX-2, iNOS, and inflammatory cytokines (P<0.05 or P<0.01). FEA also reduced NF-κB p65 translocation into the nucleus and decreased MAPK phosphorylation, indicating that the NF-κB/MAPK signaling pathway may be closely related to the inflammatory response during viral infection. The findings suggested the potential pharmaceutical application of FEA as a natural product that can treat viral infections due to its ability to mitigate inflammatory responses.     

蜂胶抗炎功效成分研究

为确定中国杨属型蜂胶的抗炎功效成分,用100ng/ml脂多糖(LPS)和40μg/ml氧化型低密度脂蛋白(ox-LDL)分别诱导脐静脉血管内皮细胞(HUVECs)损伤。通过SRB法分别检测蜂胶水提物中分离的11种组分和蜂胶乙酸乙酯提取物中分离的8段组分对2种炎症细胞模型细胞存活率的影响。结果表明:蜂胶水提物中的11种组分不同程度的促进2种炎症细胞的细胞存活率,乙酸乙酯提取物中的8段组分也是不同程度的提高2种炎症细胞的细胞存活率。由此可知,蜂胶的抗炎功效成分十分复杂,很可能是多种成分协同功效的结果,而且酚酸类物质发挥了一定的作用。     

用CODEHOP设计简并引物克隆反刍月形单胞菌K6乙酸激酶基因片段

利用CODEHOP设计细菌乙酸激酶的简并引物,选用1对引物ACKSe以高效丙酸生成菌反刍月形单胞菌K6基因组DNA进行PCR,得到749 bp PCR产物,产物经pMD18-T载体克隆转化至DH5α大肠杆菌中,测序后经Blastx比对,此DNA产物与其他菌属来源的乙酸激酶蛋白序列具有相似性,所克隆的序列即为K6的乙酸激酶基因片段。用CODEHOP程序化设计的简并引物可信性强,阳性率高。该基因的成功克隆为丙酸生成菌K6乙酸代谢工程研究提供了依据。     

乙酸浓度对奶牛乳腺上皮细胞甘油三酯含量及瘦素和过氧化物酶增殖物激活受体γ基因表达量的影响

本试验旨在研究乙酸浓度对奶牛乳腺上皮细胞甘油三酯(TAG)含量及瘦素(leptin)和过氧化物酶增殖物激活受体γ(PPARγ)基因表达量的影响。将传至第3代的奶牛乳腺上皮细胞以适宜的密度培养48 h后,随机分为6个处理,各处理分别采用乙酸浓度为0(对照组)、4、6、8、10和12 mmol/L的培养液。置于37℃5%CO2培养箱继续培养48 h。收集细胞,观测脂滴形成状况,测定TAG含量及leptin和PPARγ基因表达量。结果表明:随乙酸浓度的增加,培养的奶牛脂肪前体细胞脂滴形成增多;随乙酸浓度的增加,TAG含量和PPARγ基因表达量均呈显著的线性或二次曲线增加(P<0.05),且以乙酸浓度为10和12 mmol/L时效果较好;一定浓度的乙酸可上调leptin基因表达量,尤以浓度为8、10 mmol/L时上调作用较好。结果提示,乙酸对奶牛乳腺上皮细胞内脂滴的形成、TAG的积累、leptin和PPARγ基因的表达有显著的促进作用。本试验条件下,乙酸浓度为10～12 mmol/L时能较好地促进PPARγ基因的表达,浓度为8～10 mmol/L时能较好地促进leptin基因的表达。