分相萃取法纯化朝鲜蓟中洋蓟素的工艺研究

以朝鲜蓟提取物为原料、乙酸乙酯为萃取剂,考查萃取时间、分相时间、提取物质量浓度、相比、萃取温度、pH值等因素对萃取率的影响,并通过正交试验优化洋蓟素的最佳萃取参数。结果表明,朝鲜蓟中洋蓟素的最佳萃取参数为萃取时间30 min,分相时间80 min,提取物质量浓度10.0 g/L,相比1.25∶1,萃取温度35℃,pH值4.0,此时洋蓟素萃取率达到67.37%。     

ROCK promotes high glucose-induced cardiomyocyte apoptosis by inhi-biting PI3K/Akt signaling pathway

AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway， thus promoting the apoptosis of cardiomyocytes.     

无溶剂法制备丙烯海松酸蔗糖酯及其在空气-水界面吸附行为研究

在无溶剂条件下,以丙烯海松酸和蔗糖为原料,在高温熔融状态下通过酯化反应制备了丙烯海松酸蔗糖酯,运用红外光谱、核磁共振和凝胶渗透色谱对其结构进行了表征,并测试了其临界胶束浓度(CMC),利用再定向理论分析了丙烯海松酸蔗糖酯表面活性剂在空气-水界面的形态转变和吸附量等吸附行为。研究结果表明:成功合成了丙烯海松酸蔗糖酯,其CMC值为2.2 g/L。丙烯海松酸蔗糖酯在空气-水界面的吸附状态分为状态1和状态2,随着表面压的增加,溶剂摩尔分数逐渐降低,2种状态丙烯海松酸蔗糖酯分子在空气-水界面上的摩尔分数之和逐渐增加。计算了吸附在界面上的状态1和状态2丙烯海松酸蔗糖酯的吸附量,随着表面压增大,状态1吸附量先增大后减小,状态2吸附量占主导且持续增多,吸附量最高可达1.9 mmol/m2。丙烯海松酸蔗糖酯的吸附和胶束化摩尔自由能(ΔG)分别为-20.67和-15.16 kJ/mol,表明丙烯海松酸蔗糖酯优先吸附在界面上,达到饱和后就形成胶束。     

Glucose transport and milk secretion during manipulated plasma insulin and glucose concentrations and during LPS‐induced mastitis in dairy cows

In dairy cows, glucose is essential as energy source and substrate for milk constituents. The objective of this study was to investigate effects of long‐term manipulated glucose and insulin concentrations in combination with a LPS‐induced mastitis on mRNA abundance of glucose transporters and factors involved in milk composition. Focusing on direct effects of insulin and glucose without influence of periparturient endocrine adaptations, 18 dairy cows (28 ± 6 weeks of lactation) were randomly assigned to one of three infusion treatments for 56 h (six animals each). Treatments included a hyperinsulinemic hypoglycaemic clamp (HypoG), a hyperinsulinemic euglycaemic clamp (EuG) and a control group (NaCl). After 48 h of infusions, an intramammary challenge with LPS from E. coli was performed and infusions continued for additional 8 h. Mammary gland biopsies were taken before, at 48 (before LPS challenge) and at 56 h (after LPS challenge) of infusion, and mRNA abundance of genes involved in mammary gland metabolism was measured by RT‐qPCR. During the 48 h of infusions, mRNA abundance of glucose transporters GLUT1, 3, 4, 8, 12, SGLT1, 2) was not affected in HypoG, while they were downregulated in EuG. The mRNA abundance of alpha‐lactalbumin, insulin‐induced gene 1, κ‐casein and acetyl‐CoA carboxylase was downregulated in HypoG, but not affected in EuG. Contrary during the intramammary LPS challenge, most of the glucose transporters were downregulated in NaCl and HypoG, but not in EuG. The mRNA abundance of glucose transporters in the mammary gland seems not to be affected by a shortage of glucose, while enzymes and milk constituents directly depending on glucose as a substrate are immediately downregulated. During LPS‐induced mastitis in combination with hypoglycaemia, mammary gland metabolism was more aligned to save glucose for the immune system compared to a situation without limited glucose availability during EuG.     

Adenosine protects Sprague Dawley rats from high‐fat diet and repeated acute restraint stress‐induced intestinal inflammation and altered expression of nutrient transporters

This study investigated the effect of repeated acute restraint stress and high‐fat diet (HFD) on intestinal expression of nutrient transporters, concomitant to intestinal inflammation. The ability of adenosine to reverse any change was examined. Six‐week‐old male Sprague Dawley rats were divided into eight groups: control or non‐stressed (C), rats exposed to restraint stress for 6 h per day for 14 days (S), control rats fed with HFD (CHF) and restraint‐stressed rats fed with HFD (SHF); four additional groups received the same treatments and were also given 50 mg/l adenosine dissolved in drinking water. Fasting blood glucose, plasma insulin, adiponectin and corticosterone were measured. Intestinal expression of SLC5A1, SLC2A2, NPC1L1 and TNF‐α was analysed. Histological evaluation was conducted to observe for morphological and anatomical changes in the intestinal tissues. Results showed that HFD feeding increased glucose and insulin levels, and repeated acute restraint stress raised the corticosterone level by 22%. Exposure to both stress and HFD caused a further increase in corticosterone to 41%, while decreasing plasma adiponectin level. Restraint stress altered intestinal expression of SLC5A1, SLC2A2 and NPC1L1. These changes were enhanced in SHF rats. Adenosine was found to alleviate HFD‐induced increase in glucose and insulin levels, suppress elevation of corticosterone in S rats and improve the altered nutrient transporters expression profiles. It also prevented upregulation of TNF‐α in the intestine of SHF rats. In summary, a combination of stress and HFD exaggerated stress‐ and HFD‐induced pathophysiological changes in the intestine, and biochemical parameters related to obesity. Adenosine attenuated the elevation of corticosterone and altered expression of SLC5A1, NPC1L1 and TNF‐α.     

不同代谢葡萄糖水平对绵羊人工瘤胃发酵特性、微生物蛋白质浓度和产气参数的影响

本试验旨在通过人工瘤胃体外培养,研究不同代谢葡萄糖水平下的绵羊瘤胃发酵特性、微生物蛋白质浓度和产气参数。采用单因子试验设计,共设计4个代谢葡萄糖水平[125(A)、138(B)、153(C)、168 g/kg(D)]。体外试验所用瘤胃液采自4只安装有永久性瘤胃瘘管的绵羊。分别于培养0、2、4、6、8、12、24 h采集2 mL培养液用于分析。结果表明:1)8~24 h培养液pH随着代谢葡萄糖水平的提高而出现显著或极显著下降(P0.05或P0.01);氨氮浓度在2 h时D组显著高于A组(P0.05),而在6 h时A组显著高于其他3组(P0.05)。2)D组6h培养液细菌蛋白浓度显著高于A组(P0.05);随着代谢葡萄糖水平的提高,培养液丙酸、丁酸、总挥发性脂肪酸的浓度总体呈升高趋势,乙酸/丙酸呈下降趋势。3)随着代谢葡萄糖水平的提高,理论最大产气量极显著降低(P0.01),达1/2理论最大产气量的时间极显著缩短(P0.01);潜在产气量无显著变化(P0.05),24 h产气量和产气速率常数分别显著或极显著下降和上升(P0.05或P0.01)。结果提示,提高代谢葡萄糖水平,可以提高丙酸、总挥发性脂肪酸的浓度,同时可为绵羊提供较多的生糖前体物质。     

EMS和60^Co-γ射线辐照复合诱变黄麻突变体苗期生理生化特性研究

为丰富黄麻种质资源和扩大突变体容量,本研究采用物理诱变（60^Co-γ）和化学诱变（EMS）相结合的方法,以＂中黄麻4号＂等7个黄麻品种为材料,观察记录诱变黄麻苗期的形态学变化,分析诱变黄麻的代谢特性和抗性指标。结果表明EMS和60^Co-γ射线对黄麻幼苗的叶片产生不同程度的损伤,前者主要导致叶片卷曲,后者导致叶片分叉。复合诱变导致黄麻幼苗脯氨酸含量、丙二醛含量和根系活力提高。而可溶性蛋白、SOD活性和POD活性变化则品种之间呈现差异。本研究为黄麻种质创新的方法提供了参考,并且丰富了黄麻突变体的材料。     

温阳复方对肥胖大鼠模型糖代谢的影响

为考察温阳为主中药复方对肥胖大鼠糖代谢的影响，采用高脂饲料喂养12周建立肥胖大鼠模型，将造模成功的 SD 肥胖大鼠随机分成3组，分别为模型组、二甲双胍组、温阳复方组，并设正常对照组。连续灌胃8周，在给药第4、8周测其空腹血糖，给药 8周后测口服糖耐量与肌糖原、肝糖原、糖化血红蛋白含量。结果显示，给药 8周后，温阳复方组空腹血糖和糖化血红蛋白含量降低，肌糖原、肝糖原含量升高。结果表明，温阳复方能够明显纠正肥胖大鼠糖代谢紊乱，有效预防和延缓早期糖尿病的发生。     

Effects of phytosterol ester on aortic aging and expression of related genes in rats

AIM: To explore the protective effect of phytosterol ester (PSE) on aortic aging in rats. METHODS: The female SD rats (12 months old, n=42) were randomly divided into control group, model group and PSE group. During the experiment, the rats in control group, model group and PSE group were treated with basic feed, high-fat diet (HFD) and HFD with 2% PSE (W/W) for 6 months, respectively. The morphological changes of the aorta were observed by HE staining and Masson staining, and the absolute area of smooth muscle cells and collagen fiber in the vascular wall were measured by image analysis. The levels of advanced glycosylation end products (AGEs), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in the plasma were detected. The expression of silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor γ (PPARγ) at mRNA and protein levels in the vascular tissue was determined by real time PCR and Western blot, respectively. RESULTS: PSE significantly lowered plasma TC and LDL-C, and increased plasma HDL-C level (P<0.05), but had no effect on plasma TG level. PSE significantly attenuated the thickening of intima and media of aging aortic, and decreased the migration of vascular smooth muscle cells (VSMC) and the amount of VSMC and collagen fiber in the aorta (P<0.05). PSE significantly reduced the contents of AGEs and MDA (P<0.05), but had no effect on the activity of SOD and CAT in the plasma. PSE also down-regulated the expression of PPARγ and up-regulated the expression of SIRT1 (P<0.05). CONCLUSION: PSE is able to attenuate the senescence process in the aorta by reducing the production of reactive oxygen species in plasma, and activating SIRT1, or inhibiting the expression of PPARγ in vascular tissues.