Butylated hydroxyanisole promotes expression of neural specific genes in mouse fetal liver cells through extracellular signal-regulated kinase signaling pathways

AIM: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver Sca-1+ cells in vitro. METHODS: Sca-1+ cells from 14.5-day-old mouse fetal liver were isolated with a magnetic cell sorting kit. Cultured cells pretreated with or without extracellular signal-regulated kinase (MEK1) inhibitor, PD98059, were induced by 200 μmol/L butylated hydroxyanisole (BHA) for 24 hours, and then incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting and RT-PCR. RESULTS: There was low level of neuronfilament-L (NF-L) and brain factor-1 (BF-1) in fetal liver Sca-1+ cells, but no neuronfilament-H (NF-H) and tyrosine hydroxylase (TH) was observed. BHA significantly promoted the expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver Sca-1+ cells. NF-L, NF-H, BF-1 and TH increased by 6.32 fold, 2.73 fold, 3.37 fold and 2.68 fold, respectively (all P<0.01 vs untreated cells). However, the expression of BHA-induced genes were inhibited by ERK kinase inhibitor PD98059 (25 μmol/L). CONCLUSION: BHA induces neural differentiation of fetal liver cells through ERK.     

Effect of supplementing vitamin E analogues on post-thaw semen parameters and fertility in chicken

1. The effect of supplementing water-soluble vitamin E analogues 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (trolox) and butylated hydroxy toluene (BHT) was studied in two separate experiments.

2. In the first experiment, trolox was supplemented at 0.2 mM, 0.4 mM and 0.8 mM concentrations along with N-methylacetamide (MA; 12% final concentration) and semen was cryopreserved in 0.5 ml French straws. Different semen parameters and fertility were assessed from post-thaw samples.

3. Sperm motility, live sperm, and mitochondrial activity were significantly lower (P < 0.05) in cryopreserved semen. Lipid peroxidation (LPO) was significantly higher (P < 0.05) in cryopreserved semen that was reduced by trolox supplementation. The treatment containing trolox at 0.2 mM concentration produced significantly higher (P < 0.05) fertility compared to unsupplemented cryopreservation treatment.

4. In the second experiment, BHT was supplemented at 0.25 mM, 0.5 mM, and 1 mM concentrations along with MA during semen cryopreservation.

5. Sperm motility, live sperm and MTT dye reduction test were significantly lower (P < 0.05) in cryopreserved semen. These parameters declined with increasing BHT concentration. Abnormal sperm was significantly higher (P < 0.05) in the BHT supplemented treatments. The sperm chromatin dispersion (SCD) test was significantly higher (P < 0.05) in cryopreserved samples and was highest in samples supplemented with 0.5 mM and 1 mM BHT. The percentage fertility was significantly lower (P < 0.05) in cryopreserved semen and BHT supplementation did not improve fertility.

6. In conclusion, trolox supplementation at 0.2 mM concentration during semen cryopreservation improved fertility, whereas BHT supplementation resulted in a decline in post-thaw semen parameters.     

二丁基羟基甲苯对雨生红球藻虾青素和油脂积累的影响

以雨生红球藻(Haematococcus pluvialis)为对象,研究高光照、缺氮条件下,外源添加不同浓度的二丁基羟基甲苯(Butylated hydroxytoluene, BHT)对雨生红球藻生长、虾青素积累、油脂合成、脂肪酸组成、碳水化合物、蛋白含量以及虾青素和脂肪酸合成相关酶基因的影响。结果显示,添加不同浓度的BHT后,2 mg/L BHT添加组虾青素积累量为最高,显著高于其他实验组和对照组(P<0.05),达到31.66 mg/g,是对照组的1.87倍。油脂含量达45.56%,高于同期对照组(39.06%),脂肪酸组成变化不显著。在此条件下,虾青素合成关键酶基因dxs和bkt表达水平分别是对照组的5.19倍和2.04倍;脂肪酸合成关键酶基因kas和acp表达水平较对照组显著提高(P<0.05),分别是对照组的4.56倍和3.02倍。与对照组相比,2 mg/L BHT添加组的碳水化合物和蛋白含量均呈下降趋势。研究表明,在胁迫条件下,外源添加适量浓度的BHT能有效促进雨生红球藻中虾青素的积累,同时提高了藻细胞内的油脂含量。     

Chemical profiling and biological screening of Thymus lotocephalus extracts obtained by supercritical fluid extraction and hydrodistillation

Essential oil and extracts from the aerial parts of Thymus lotocephalus were obtained by hydrodistillation (HD) and supercritical fluid extraction (SFE) in two different collectors, respectively. SFE was conducted at 40 °C and a working pressure of 12 or 18 MPa. The chemical profiles were determined using GC-FID and GC-IT-MS. Oxygen-containing monoterpenes were the primary constituents in the essential oil and SFE extracts collected in the second separator, while the extracts obtained in the first separator were predominantly oxygen-containing sesquiterpenes. A large number of compounds were identified by hydrodistillation and, in contrast, the highest extraction yields were obtained using SFE. Linalool (10.43 ± 1.63%) was the main component in essential oil, whereas camphor (7.91 ± 0.84%) and cis-linalool oxide (7.25 ± 1.45%) were the major compounds in the extracts-2nd separator obtained at pressures of 12 and 18 MPa, respectively. Caryophyllene oxide was the primary constituent identified in the extracts-1st separator (4.34 ± 0.51 and 4.41 ± 1.25% obtained at 12 and 18 MPa, respectively). The antioxidant activity was assessed by ORAC and DPPH assays, and the anti-cholinesterase activity was evaluated in vitro using Ellman's method. The essential oil and SFE extracts (first separator) of T. lotocephalus possessed antioxidant activity and strongly inhibited cholinesterases. We also demonstrated that the acetylcholinesterase and butyrylcholinesterase inhibitory activities of the essential oil could be attributed to 1,8-cineole and caryophyllene oxide, respectively.     

抗氧化剂BHT微胶囊化产品超微结构的测定

用扫描电子显微镜(SEM)研究抗氧化剂二丁基羟基甲苯(BHT)微胶囊化产品的超微结构。观察了喷雾干燥法不同壁材组成和工艺条件下制作的微胶囊化产品的表面结构和内部结构,同时考察了喷雾干燥进风温度对微胶囊化产品膜结构的影响。结果表明,不同壁材组成和工艺条件下制作的微胶囊化产品的超微结构有较大差异,合适的壁材组成和工艺条件有利于提高产品质量,同时喷雾干燥进风温度对微胶囊化产品的结构有很大影响,从而也影响到产品的质量。微胶囊化产品超微结构的研究为分析影响产品质量的因素提供了重要依据。     

Evaluation of antioxidant activities of the edible and medicinal Suaeda species and related phenolic compounds

Antioxidants are the chemical substances that reduce or prevent oxidation. The present study aimed to assess in vitro and ex vivo antioxidant activities of four acetonic extracts Tunisian halophytes (Suaeda fruticosa, Suaeda pruinosa, Suaeda mollis and Suaeda maritima). Various experimental models were used for characterization of antioxidant activities of shoot extracts. Eventually, the promising specie was subjected to phenolic identification using RP-HPLC. The analyzed shoot extracts exhibited that antioxidant activities varied considerably as function of species. The highest DPPH scavenging ability was found in S. mollis with the lowest IC₅₀ value (2.5 μg/ml), followed by S. pruinosa, S. fruticosa and S. maritima. The same tendency was observed with ferric reducing power. Concerning β-carotene bleaching assays and total antioxidant activity, results showed that S. fruticosa exhibited the highest antioxidant ability against the inhibition of β-carotene bleaching, and a better total antioxidant capacity. Moreover antioxidant capacities using ORAC method and a cell based-assay showed that S. mollis, S. fruticosa, and S. pruinosa exhibit statistically similar antioxidant activity. The identification of phenolic compounds in S. mollis extract using RP-HPLC revealed that 2,5-dihydroxybenzoic acid and rutin hydrate were the major molecules. These results suggested that Suaeda species showed a variability of their antioxidant activities.     

Effect of plant age and temperature on bolting in Chinese cabbage

The antioxidant BHT — commonly used as a food stabilizer — when applied in aqueous solution to the tips of 5-day-old pea seedlings, to young apple seedlings or to germinating lettuce seeds, produced a suden cessation of growth that, when growth was resumed, was followed by certain morphological changes. Treatments with BHT caused the formation of (1) a bunch of new shoots arising at the apex of apple seedlings, (2) 2 or 3 adventitious fast-growing shoots from the base of the pea hypocotyl, and (3) adventitious growing points in lettuce plants. This multiplication of growing points was followed, during further development, by splitting of the apple seedling stems and lettuce leaf blades into independently growing organs.