一株产蜜柚果汁脱苦酶的青霉菌的筛选及初步鉴定

对前期分离获得的柚苷酶产生真菌进行复筛，从29株脱苦酶产生真菌中复筛出了5株脱苦能力较强的真菌，其中实验室编号为3-3-21的菌株对柚皮苷的分解率为70%，对柠碱和诺米林的分解率分别为27%和14%，表明其具有较强的脱除蜜柚果汁苦味的能力。该株菌初步鉴定为常现青霉组(P.frequentans)。     

In ovo injection of flavanone on bone quality characteristics,biochemical parameters and antioxidant enzyme status of blood in daily chicks

In ovo injection (IOI) of Naringin (N), flavanone was examined on post‐hatch blood biochemical parameters, antioxidant status and bone characteristics. Fertile eggs (n = 700) were distributed in seven groups with 100 eggs. On 14th and 17.5th days of incubation, four groups were injected using 15 or 30 mg active ingredient levels of naringin/0.5 ml saline/egg, low and high level, into amnion sac. Controls include sham (injected normal saline, 0.5 ml/egg on day 14 and 17.5th) and un‐injected group. IOI of high naringin and saline on 14th day of incubation resulted in lower hatchability and then higher mortality in last week of embryonic life. On day hatch, high levels of injected groups more body weight compared to the control. Chick length was increased at high levels of naringin on day 17.5th compared to control and saline injected. Quality traits of bones were improved in naringin‐injected groups compared to control. IOI of naringin influenced thyroid hormones on 14th day of incubation. Naringin groups influenced the Alkaline phosphatase (ALP), Calcium (Ca), superoxide dismutase (SOD), blood biochemical and lipids. Totally, amniotic IOI of naringin in last days of developing embryo may be useful for hatched chick, development of leg long bone or effect on biochemical metabolites by levels of flavanone that it needs more research.     

超声辅助响应面法优化种植佛手中橙皮苷和柚皮苷提取工艺的研究

为了优化提取万州种植佛手中橙皮苷和柚皮苷的工艺条件,采用响应面法优化超声辅助进行研究。在超声提取时间、乙醇体积、超声提取温度、液固比单因素试验基础上,研究了超声条件下影响万州种植佛手中橙皮苷和柚皮苷含量的几个因素,采用Box-Behnken试验设计和响应面分析法考察前述4个条件对万州种植佛手中橙皮苷和新橙皮苷含量的影响,采用RP-HPLC法测定万州种植佛手中橙皮苷和柚皮苷的含量。结果表明,最佳超声辅助提取工艺为超声提取时间26 min、乙醇体积分数82%、超声提取温度62℃、佛手与乙醇的质量比1:22 (g/mL),万州种植佛手中橙皮苷和柚皮苷含量分别为0.072%±0.315%和0.082%±0.753%,与预测值相符。本研究对万州种植佛手超声辅助提取的工艺可靠性高,有效成分提取率高,适合于工业化生产推广。     

The effects of naringenin and naringin on the glucose uptake and AMPK phosphorylation in high glucose treated HepG2 cells

BackgroundNaringin and its aglycone naringenin are citrus-derived flavonoids with several pharmacological effects. On the other hand, the mechanism for the anti-diabetic effects of naringenin and naringin are controversial and remain to be clarified further.ObjectiveThis study examined the relationship between glucose uptake and AMP-activated protein kinase (AMPK) phosphorylation by naringenin and naringin in high glucose-treated HepG2 cells.MethodsGlucose uptake was measured using the 2-NBDG fluorescent D-glucose analog. The phosphorylation levels of AMPK and GSK3β (Glycogen synthase kinase 3 beta) were observed by Western blotting. Molecular docking analysis was performed to evaluate the binding affinity of naringenin and naringin to the γ-subunit of AMPK.ResultsThe treatment with naringenin and naringin stimulated glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells. Both flavonoids increased glucose uptake by promoting the phosphorylation of AMPK at Thr172 and increased the phosphorylation of GSK3β. Molecular docking analysis showed that both naringenin and naringin bind to the γ-subunit of AMPK with high binding affinities. In particular, naringin showed higher binding affinity than the true modulator, AMP with all three CBS domains (CBS1, 3, and 4) in the γ-subunit of AMPK. Therefore, both naringenin and naringin could be positive modulators of AMPK activation, which enhance glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells.ConclusionsThe increased phosphorylation of AMPK at Thr172 by naringenin and naringin might enhance glucose uptake regardless of insulin stimulation in high glucose treated HepG2 cells.     

Influence of citrus flavonoids on laying hen performance,inflammatory immune response,egg quality and yolk oxidative stability

1. The objective of this study was to investigate the effects of dietary supplementation with natural flavonoids (naringin and hesperidin) on laying hens’ performance, cellular immunity and egg quality parameters.

2. A total of 72 individually caged laying hens were allocated into 1 of 6 treatment groups: a control (C) group that was fed with a basal diet and groups that were offered the same diet further supplemented with hesperidin at 0.75 g/kg (E1), or 1.5 g/kg (E2), or naringin at 0.75 g/kg (N1), or 1.5 g/kg (N2) or α-tocopheryl acetate at 0.2 mg/kg (VE) for 67 d.

3. Supplementation with naringin or hesperidin did not affect the performance and egg quality (P > 0.05) apart from an improvement in the yolk colour that was more orange in naringin and hesperidin groups in comparison to the controls (P-linear < 0.05). Egg yolk and plasma cholesterol levels were not affected by citrus flavonoids (P-linear > 0.05).

4. Inflammatory immune response, measured by phytohaemagglutinin skin test (PHA), was suppressed in laying hens that were fed with either naringin (P-linear < 0.05) or hesperidin (P < 0.05). Egg yolk oxidative stability was improved from the 4th d after naringin or hesperidin supplementation. This beneficial effect was comparable to that of α-tocopheryl acetate and was observed in eggs that were stored for up to 120 d.

5. In conclusion, naringin and hesperidin may favourably prolong the shelf life of eggs, appear to possess anti-inflammatory properties and could improve the yolk colour without any side effects on the performance or egg quality traits.     

高效液相色谱法测定乳与乳制品中柚皮苷

建立高效液相色谱法测定乳与乳制品中柚皮苷含量的方法。样品用甲醇提取,在284 nm波长下,流动相为体积分数0.1%醋酸水溶液-乙腈（75∶25,V/V）,C18色谱柱分离,经二极管阵列检测器检测,外标法定量。结果表明：柚皮苷在5～500 μg/mL质量浓度范围内线性关系良好,相关系数为0.999 9;分别向乳粉、液态乳和酸乳样品中在加入柚皮苷标准品0.02、0.04、0.08 g/100 g时,加标回收率分别为96.2%～103.2%、96.3%～103.7%和99.7%～104.1%,相对标准偏差分别为1.12%～2.31%、1.25%～1.85%和1.24%～1.66%。该检测方法简便、快速、准确、重复性好,能够测定乳与乳制品中柚皮苷的含量。     

柚皮甙、橙皮甙和儿茶素硫酸酯的合成及其HPLC-MS的鉴定

目的合成柚皮甙、橙皮甙、儿茶素硫酸酯。方法柚皮甙、橙皮甙、儿茶素在冰浴中与浓硫酸反应2 h,用6N NaOH终止反应。衍生物用高效液相色谱(HPLC)-大气压化学电离(APCI)-质谱(MS)联用鉴定。结果和结论有柚皮甙、橙皮甙和儿茶素硫酸酯的生成。     

Restoration of the adipogenic gene expression by naringenin and naringin in 3T3-L1 adipocytes

BackgroundNaringenin and its glycoside naringin are well known citrus flavonoids with several therapeutic benefits. Although the anti-adipogenic effects of naringenin and naringin have been reported previously, the detailed mechanism underlying their anti-adipogenesis effects is poorly understood.ObjectivesThis study examined the anti-adipogenic effects of naringenin and naringin by determining differential gene expression patterns in these flavonoids-treated 3T3-L1 adipocytes.MethodsLipid accumulation and triglyceride (TG) content were determined by Oil red O staining and TG assay. Glucose uptake was measured using a 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose fluorescent d-glucose analog. The phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl Co-A carboxylase (ACC) were observed via Western blot analysis. Differential gene expressions in 3T3-L1 adipocytes were evaluated via RNA sequencing analysis.ResultsNaringenin and naringin inhibited both lipid accumulation and TG content, increased phosphorylation levels of both AMPK and ACC and decreased the expression level of 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in 3T3-L1 adipocytes. RNA sequencing analysis revealed that 32 up-regulated (> 2-fold) and 17 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Scd1, Mogat1, Dgat, Lipin1, Cpt1a, and Lepr, were normalized to the control level in naringenin-treated adipocytes. In addition, 25 up-regulated (> 2-fold) and 25 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Fabp5, Scd1, Srebf1, Hmgcs1, Cpt1c, Lepr, and Lrp1, were normalized to the control level by naringin.ConclusionsThe results indicate that naringenin and naringin have anti-adipogenic potentials that are achieved by normalizing the expression levels of lipid metabolism-related genes that were perturbed in differentiated 3T3-L1 cells.     

贮藏温度对柑橘果肉中柠檬苦素和柚皮苷含量变化的影响

将国庆1号和国庆3号温州蜜柑的成熟果实分别贮藏在4℃和常温下,利用高效液相色谱法检测贮藏过程中果肉主要苦味物质的含量变化。结果表明:2个品种的成熟果肉中检测不到诺米林,采后0d柚皮苷含量为柠檬苦素的10倍以上;在4℃和常温贮藏时,柠檬苦素和柚皮苷含量总体上均呈增加的趋势,有一定波动,但冷库贮藏时变化较剧烈;国庆1号受低温影响较大,使其在几个时期的2种苦味物质含量发生了显著变化;4℃低温贮藏利于苦味物质的增加,柠檬苦素尤甚,而贮后0～20d是其敏感时期。     

Comparison of the effects of dietary supplementation of flavonoids on laying hen performance,egg quality and egg nutrient profile

1. The aim of this experiment was to compare the effects of dietary supplementation of hesperidin, naringin and quercetin on laying hen performance, egg quality and egg yolk lipid and protein profiles.

2. A total of 96 Lohmann White laying hens weighing an average of 1500 g at 28 weeks of age were randomly assigned to a basal diet and the basal diet supplemented (0.5 g/kg) with either hesperidin, naringin or quercetin. Each treatment was replicated in 6 cages in an 8-week experimental period. Data were analysed using one-way analysis of variance.

3. None of the dietary flavonoids affected laying performance and eggshell quality. Hesperidin and quercetin supplementations decreased albumen and yolk indexes.

4. As compared to the control group, egg yolk cholesterol content decreased and egg yolk protein content increased in response to dietary hesperidin and quercetin supplementation. The mean egg yolk cholesterol (mg/g) and protein (g/100 g) contents were 10.08/14.28, 16.12/14.08, 14.75/15.04 and 15.15/14.85 for the control group and groups supplemented with naringin, hesperidin and quercetin, respectively.

5. Egg yolk lipid and protein profiles were variable.

6. In conclusion, dietary supplementation of hesperidin or quercetin could be used in the diets during the early laying period to reduce egg yolk cholesterol and increase egg yolk protein, which may be attractive to consumers.