Ellagic acid attenuates hypoxic-ischemic brain injury by alleviating autophagy

AIM:To investigate whether ellagic acid (EA) attenuates hypoxic-ischemic encephalopathy (HIE) by down-regulating autophagy. METHODS:In vivo, Sprague-Dawley rats (n=17) were randomly divided into 3 groups:5 rats for sham group, 6 rats for HIE group and 6 rats for HIE+EA pretreatment group. The rats in HIE+EA pretreatment group were treated with EA (10 mg/kg, 10 mL/kg, suspended in corn oil, ig). After 24 h of operation, the rats from each group were sacrificed and their brains were collected. TTC staining and HE staining were used to define the infarct areas and brain structure. The autophagy-related proteins beclin-1, P62, LC3-Ⅱ/-I and Atg5 in the cortex in each group were compared by Western blot. In vitro, PC12 cells were divided into 3 groups:control group, CoCl₂ group and CoCl₂+EA pretreatment group. CoCl₂ at 800 μmol/L was added to the PC12 cells to induce an anoxic environment. The PC12 cells were pretreated with EA at 8 μmol/L and the cell viability was measured by CCK-8 assay. The production of reactive oxidative species (ROS) in the cells was detected by flow cytometry with DCFH-DA staining. MDC staining and TMRE staining were applied to reflect the extent of autophagy and the state of apoptosis, respectively. The autophagy-related proteins in PC12 cells were also investigated. RESULTS:In HIE group, 7-day-old rats were given the operations and the their large infarct areas in the hemisphere were observed by TTC staining. HE staining displayed the injured hemispheres which contained few neurons, and exhibited edema status and serious structural damage. EA pretreatment decreased the infarct area and alleviated the damage to hemisphere with more visible neurons, compared with HIE group. Compared with sham group, the levels of autophagy-related proteins Atg5, beclin-1 and LC3-Ⅱ/-I in the cortex were increased (P<0.01), and P62 protein expression was decreased (P<0.01) in HIE group. Compared with HIE group, the protein expression of Atg5, beclin-1 and LC3-Ⅱ/-I was decreased (P<0.01) and P62 protein expression was increased in HIE+EA pretreatment group (P<0.01). In vitro, compared with CoCl₂ group, the PC12 cells in CoCl₂+EA pretreatment group showed a lower ROS level. Moreover, the cells in CoCl₂+EA pretreatment group exhibited higher mitochondrial membrane potential than that in CoCl₂ group. MDC staining in CoCl₂ group showed high value of fluorescence and increased number of autophagosomes. EA pretreatment reduced the number of autophagosomes and the extent of autophagy to protect PC12 cells. Furthermore, the protein levels of Atg5, beclin-1 and LC3-Ⅱ/-I in CoCl₂ group were higher (P<0.01), and the protein expression of P62 was lower (P<0.01) than those in control group. In CoCl₂+EA pretreatment group, the protein levels of Atg5, beclin-1 and LC3-Ⅱ/-I were decreased (P<0.01) and the protein expression of P62 was increased as compared with CoCl₂ group (P<0.01). CONCLUSION:EA pretreatment attenuates autophagy to protect the neurons against HIE injury.     

Crude protein in low‐fish meal diets for juvenile Litopenaeus vannamei can be reduced through a well‐balanced supplementation of essential amino acids

The use of supplemental essential amino acids (EAAs) has been shown to provide an opportunity to minimize excess levels of crude protein (CP) in animal feeds. The present study investigated the effect of reducing the amount of CP in low‐fish meal diets (5%) for juvenile Litopenaeus vannamei. Four sets of diets were prepared containing (% on a fed basis, mean ± SD) 31.24 ± 0.71, 33.70 ± 0.41, 36.90 ± 0.44, and 39.63 ± 0.14% CP, with each protein level containing a total dietary methionine (Met) (Met + Cys) level of 0.56 ± 0.02 (1.07 ± 0.23), 0.71 ± 0.01 (1.22 ± 0.20), 0.88 ± 0.02 (1.38 ± 0.22), or 1.04 ± 0.02% (1.55 ± 0.18%). Shrimp of 1.00 ± 0.08 g were stocked in 84 outdoor tanks of 1 m³ at a rate of 100 shrimp/m² and raised for 75 days. Final survival ranged from 83 to 97% and was unaffected by Met content. Both survival and yield were significantly depressed when shrimp were fed the 31% CP diet. Shrimp grew at a weekly rate of between 0.79 and 0.97 g, achieving a final body weight (BW) in excess of 10.8 g. There was a significant interaction between CP and Met over BW. Shrimp fed 0.56% Met achieved the lowest BW at harvest. Increasing CP beyond 34% did not enhance BW. With a dietary Met content of 0.71%, the highest BW was achieved with 34% CP compared to other levels. There was a significant improvement in Feed Conversion Ratio (FCR) when CP was raised from 31 to 34%. Similarly, dietary Met levels above 0.71% resulted in a significantly better FCR compared to 0.56%. Our study has shown that, if dietary Met (Met + Cys) meets a minimum of 0.71% (1.22%), levels of CP could be reduced from 40 to 34% without adverse effects on shrimp performance.     

氨基酸配合硼喷施对油麦菜硼营养及生长、品质的影响

为探究有机营养类物质氨基酸与硼酸配合喷施对油麦菜生长的应用效果,本研究采用盆栽试验,设置单独喷施1%硼酸处理,以及谷氨酸、丙氨酸和天冬氨酸3种氨基酸各设3个喷施浓度(5、10、20 mmol·L^-1) 与1%硼酸配合喷施处理,并以喷施清水为对照(CK),共11个处理,收获后测定植株的生长指标和硼含量。结果表明,单独喷施硼酸对油麦菜生物量无明显影响,低浓度氨基酸与硼配合喷施均能显著提高油麦菜生物量,其中丙氨酸-硼喷施处理油麦菜生物量较CK平均增加了9.4%。与CK相比,3种氨基酸与硼配合喷施均能明显提高油麦菜叶片总蛋白含量,同时降低硝酸盐积累,从而改善品质。与单独喷施硼酸相比,适宜浓度氨基酸与硼配合喷施均可显著提高油麦菜氮、钾积累量,另外谷氨酸-硼和天冬氨酸-硼喷施对油麦菜磷积累量也有明显促进作用。与单独喷施硼酸相比,氨基酸与硼配合喷施均可显著提高油麦菜地上部硼含量,其中丙氨酸-硼喷施处理油麦菜地上部硼含量平均增幅为41.0%;此外,丙氨酸-硼喷施对油麦菜地下部硼含量也有明显提升作用,平均增幅为15.6%;喷施丙氨酸浓度与油麦菜地上部和地下部硼含量均呈显著正相关关系。综合比较不同氨基酸与硼酸配合喷施对油麦菜的影响,可考虑将丙氨酸作为增效剂与硼肥配合施用,提高硼肥的应用效应。本研究为开发新型硼肥提供了理论依据。     

草酸处理对桂七芒果冷害及细胞壁代谢的影响

为探讨草酸处理对低温冷藏下芒果冷害及细胞壁代谢的影响,本试验以桂七芒果果实为试材,采用5 mmol·L^-1草酸溶液浸泡处理,以清水浸泡处理为对照,并于4℃贮藏,分析芒果的冷害指数、丙二醛(MDA)含量、相对电导率、硬度、细胞壁物质含量、细胞壁代谢酶的变化。结果表明,与对照组相比,草酸处理显著降低了低温贮藏14 d后桂七芒果果实冷害指数、MDA含量、相对电导率、原果胶和纤维素含量,显著降低了低温贮藏28 d后果实的硬度;显著提高了低温贮藏14 d后果实水溶性果胶含量及多聚半乳糖醛酸酶(PG)、果胶甲酯酶(PME)、纤维素酶(Cx)活性,显著提高了贮藏21 d后果实β-半乳糖苷酶(β-Gal)活性。综上所述,草酸处理能减轻桂七芒果冷害,维持采后果实细胞壁降解酶较高活性和水溶性果胶含量。本研究为揭示草酸减轻芒果果实冷害机制提供了依据,可为草酸应用于其他冷敏型果实的贮藏保鲜提供理论参考。     

镉和模拟酸雨胁迫对苦楝幼苗镉含量及叶片抗氧化系统的影响

为探讨苦楝在重金属Cd和酸雨的单一及复合胁迫下的生理响应特征,采用人工配置土壤盆栽试验,研究不同浓度Cd(0、30、60、90、120、150、180 mg·kg^-1)和3个梯度模拟酸雨(pH值5.6、4.5、3.5)单一及复合处理对苦楝幼苗生物量、Cd含量及叶片保护性酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)]活性和抗坏血酸-谷胱甘肽循环(AsA-GSH)的影响。结果表明,除低浓度Cd(≤30 mg·kg^-1)单一及其与pH值4.5酸雨复合处理对幼苗地上部、地下部生物量有促进作用外,其他处理均表现出抑制作用,其中复合处理对苦楝幼苗的抑制作用更明显,表现为叠加效应;单一Cd处理时,随着Cd浓度的增加,幼苗根、茎、叶中Cd含量逐渐升高,且根系大于茎叶,复合处理抑制了植物对Cd的积累,对叶片的抑制作用最强;Cd和酸雨单一及复合处理下叶片SOD、CAT和POD活性均有所增加,分别在60 mg·kg^-1 Cd、pH值5.6,60 mg·kg^-1 Cd、pH值4.5和90 mg·kg^-1Cd、pH值4.5时出现阈值;在AsA-GSH循环中,单一Cd处理下,AsA、GSH含量和抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性均随着Cd浓度的增加呈先上升后下降的趋势,单一酸雨处理对上述4个指标的影响不尽一致,而复合处理下上述指标在Cd浓度≤60 mg·kg^-1、pH值≥4.5时均能保持较高值,且随着胁迫程度的加深均出现了阈值。综上所述,苦楝对重金属Cd和酸雨胁迫具有一定的耐性,可在Cd浓度≤60 mg·kg^-1和pH值≥4.5的酸雨双重污染区域正常生长。本研究结果为土壤Cd污染的酸雨区域的修复植物筛选提供了理论基础和参考依据。     

硅钙钾镁肥配施黄腐酸钾对土壤酶活性及桃幼树生长的影响

为探究硅钙钾镁肥配施黄腐酸钾对桃树生长的效应,以盆栽一年生瑞蟠21号毛桃(Amygdalus persica Linn.)为试验材料,设置3种不同施肥方式(T1:硅钙钾镁;T2:硅钙钾镁+黄腐酸钾;CK:空白对照),在桃幼树生长季进行追肥处理,研究硅钙钾镁肥及硅钙钾镁肥配施黄腐酸钾对土壤酶活性、土壤养分状况及桃幼树根系构型和植株生长的影响。结果表明,与CK相比,T1、T2均显著提高了桃幼树生长季各时期土壤酸性磷酸酶、脲酶和过氧化氢酶活性,以及土壤中碱解氮、有效磷、速效钾和有效硅等含量,且配施黄腐酸钾后作用更明显。与CK相比,T1、T2细根比重分别提高了11.2%和26.2%,根系活力分别提高了12.2%和18.1%,根系总长度分别增加了28.1%和61.9%,且有效延缓了根系衰老;T1、T2植株总干重分别提高了1.2倍和2.0倍,根冠比分别提高了9.4%和17.0%,且差异显著。与CK相比,T1、T2各器官中全氮、全磷、全钾含量均显著增加,地上部硅含量分别提高了61.3%和91.9%,地下部硅含量分别提高了62.3%和102.5%,且差异显著。同时,T1、T2桃幼树叶片叶绿素含量和净光合速率均显著提高。综上所述,硅钙钾镁肥可有效提高土壤酶活,促进桃幼树细根的生长,延缓根系衰老进程,增加氮、磷、钾、硅等营养元素积累量,提高叶片光合速率,促进植株生长;硅钙钾镁肥配施黄腐酸钾后,提高了土壤有效硅及植株硅含量,比单施硅钙钾镁肥效果更加显著。本研究结果为桃树合理施用硅钙钾镁肥提供了理论依据。     

辐照协同甲酸分离油茶壳中纤维素、木质素和木糖的工艺研究

为了提高油菜壳木质纤维素分离效率,本试验以油茶壳为原料,研究不同辐照剂量(0、200、400、600、800 kGy)处理后其粉碎能耗、粒度分布和化学组分的变化,同时开展辐照协同甲酸分离油茶壳木质纤维素的工艺研究。结果表明,辐照处理后油茶壳粉碎能耗降低,粉碎后细颗粒样品所占比例增加,经400 kGy剂量辐照处理的油茶壳粉碎能耗比对照节约43.59%,800 kGy剂量辐照处理的油茶壳粉碎后粒径<0.075 mm的颗粒占比达到41.38%。辐照处理后油茶壳木质纤维素发生降解,水溶性组分、水溶性单糖、聚糖含量均增加。辐照协同甲酸分离油茶壳纤维素、木质素和木糖的最佳工艺为:辐照剂量400 kGy,反应温度100℃,反应时间3 h,在此条件下分离获得的油茶壳纤维素、木质素、木糖的提取率分别为89.94%、47.74%和96.37%,纤维素和木质素纯度分别为45.05%和91.92%。本研究结果对油茶壳木质纤维素全组分的高效分离和应用具有重要意义。     

连作植烟土壤具有显著积累特征的两种酚酸的化感效应评价

为科学评价连作植烟土壤中具有显著积累特征的两种酚酸的化感效应,丰富现有连作障碍机理,以烤烟品种K326为试验材料,基于前期田间实测结果设置不同浓度梯度(间苯三酚:0.20、1.00、5.00 mg·kg^-1,对羟基苯甲酸0.02、0.10、0.50 mg·kg^-1)进行盆栽试验。结果表明,两种酚酸物质均显著抑制了烤烟种子萌发。间苯三酚抑制了幼苗根系的生长,且浓度越大抑制作用越强;对羟基苯甲酸对幼苗根系生长的影响表现出“低促高抑”的浓度效应。随着间苯三酚浓度的增加,叶片超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性呈上升趋势,而过氧化物酶(POD)活性呈下降趋势;随着对羟基苯甲酸浓度的增加,叶片SOD、CAT和POD活性均呈先升后降趋势。此外,两种酚酸物质均降低了烤烟幼苗叶片叶绿素含量,增加了丙二醛(MDA)含量。随着处理时间的延长,胁迫作用趋势相同。由此可知,间苯三酚和对羟基苯甲酸不同程度地抑制了烤烟幼苗的生长,且存在浓度效应。间苯三酚表现为抑制作用,浓度越高抑制作用越强;对羟基苯甲酸浓度≤0.10 mg·kg^-1时表现为促进作用,>0.10 mg·kg^-1时表现为抑制作用。对羟基苯甲酸较间苯三酚具有更强的生物毒性。本研究可以丰富植烟土壤连作障碍机理,为今后连作障碍调控提供理论依据。     

不同林龄杉木林下套种阔叶树对土壤磷组分的影响

套种是杉木人工林经营的重要措施,磷是南方森林生态系统中主要限制性养分元素之一,但套种模式对土壤磷素的影响尚不明确。以亚热带杉木人工林表层(0-10 cm)土壤为对象,研究套种林(杉阔套种幼林、杉阔套种成熟林)和杉木幼林土壤理化性质和土壤各形态磷含量差异,分析套种对杉木人工林土壤磷含量的影响。结果表明:(1)不同套种林显著改变土壤总磷、土壤总无机磷、土壤总有机磷、土壤微生物生物量磷(MBP)和土壤酸性磷酸酶活性(APA),大小顺序均为杉阔套种成熟林>杉阔套种幼林>杉木幼林。(2)土壤各磷组分中活性磷含量较低,其中NaHCO3-Po在活性组分中占主导;土壤NaOH-Po是中等活性磷的主要组分,杉阔套种成熟林尤为显著;闭蓄态磷(Residual-P)在总磷含量中最高。(3)与杉木幼林相比,杉阔套种成熟林显著增加了树脂提取态磷(Resin-Pi)、碳酸氢钠提取态有机和无机磷(NaHCO3-Pi、NaHCO3-Po)、氢氧化钠提取态有机和无机磷(NaOH-Pi、NaOH-Po)、氢氧化钠残留提取态有机磷(NaOHu.s-Po)、盐酸提取态磷(HCl-Pi)和闭蓄态磷(Residual-P)含量;土壤总无机磷、NaHCO3-Po、HCl-Pi、NaHCO3-Pi、NaOHu.s-Pi和Residual-P对杉阔套种幼林的响应不敏感。(4)除含水率外不同林龄下杉阔套种林土壤磷形态与土壤理化性质(土壤总碳氮、土壤可溶性有机氮、土壤微生物生物量磷、酸性磷酸酶)呈正相关性(P<0.05)。冗余分析表明,土壤磷组分的变化主要受MBP调控,且MBP与有机磷组分(NaOHu.s-Po、NaOH-Po)和HCI-Pi呈显著正相关。总之,套种林的土壤磷素有效性高于杉木幼林,土壤养分状况更佳。