A new alkylene dihydrofuran glycoside with antioxidation activity from the root bark of Morus alba L.

A new alkylene dihydrofuran glycoside（1） was isolated from the root bark ofMorus alba L.,along with moracin M-3′-O-β-D-glucopyranoside （2）,and moracin M-6,3′-di-O-β-D-glucopyranoside（3）.Compound 1 was identified as 2-methylene-3-methoxy-2, 5-dihydrofuran-4-O-β-D-glucopyranoside on the basis of chemical and spectroscopic data including 1D and 2D NMR spectral analysis.In addition,the antioxidant activity of 1 was evaluated using the 1,1-diphenyl-2-picrylhydrazyl（DPPH） and 2,2′-azinobis- 3-ethylbenzothiazoline-6-sulphonic acid（ABTS） assay.The IC_（50） values were 2.49 and 0.45 mg/mL,respectively.

Research and Prospect of Mulberry (Morus alba L.)

Mulberry(Morus alba L.)is not only rich in nutrients,but also has medicinal value.As a drug and food,it is widely planted all over the country,with considerable yield and economic value.In recent years,it has attracted many scholars and been widely reported,and in-depth research has already been conducted.Through reviewing the literatures on mulberry,this paper summarized the chemical composition,pharmacological effects and quality evaluation of mulberry,providing some reference for the further development and utilization of mulberry.

Evaluation of the Liver Cancer Prevention of Anthocyanin Extracts from Mulberry (<i>Morus alba</i>L.) Variety PR-01

This study aims to evaluate the preventive effects of anthocyanins extracts (MAEs) from mulberry variety PR-01 against N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in rats. It was found that 150 mg·kg-1 MAEs treatment significantly reduced the NDEA-induced hepatic nodules incidence and hepatocellular carcinoma incidence by 58.30% and 41.70% compared to the model group. Meanwhile, MAEs significantly restored the elevated the liver function enzymes, inhibited the tumor necrosis factor alpha and interleukin-6 levels, elevated the serum interleukin-10 and interferon-γ and increased hepatic glutathione-S-transferase and UDP-glucuronosyltransferase 2B1 enzyme activity. Moreover, 150 mg·kg-1 MAEs supplement enhanced glutathione content and the activities of superoxide dismutase, catalase, glutathione peroxidase activities but reduced the malondialdehyde and thiobarbituric acid-reactive substances content by 37.90% and 44.52%. Furthermore, MAEs pretreatment maintained nuclear factor erythroid 2-related factor 2 (Nrf2), Kelch-like ECH-associated protein 1, heme oxygenase-1, and NAD(P)H: quinine oxidoreductase1 stimulation and inhibited the expression of TNF-α, nuclear factor-kappaB (NF-κB), and cyclooxygenase-2 (COX-2), indicating that MAEs exhibit effectively prevention effects against liver cancer via decreased lipid peroxidation, induced Nrf2-mediated antioxidant enzymes and attenuating the inflammatory mediators COX-2 through NF-κB pathway. Thus, MAEs of mulberry variety PR-01 may be used as a good functional dietary supplement against liver cancer.

Selection and characterisation of lactic acid bacteria isolated from different origins for ensiling Robinia pseudoacacia and Morus alba L.leaves

The objective of this study was to isolate lactic acid bacteria（LAB） strains from different origins and to select the best strains for ensiling Robinia pseudoacacia（RB） and Morus alba L.（MB）leaves. The LAB strains were inoculated into the extracted liquid obtained from RB and MB leaves to evaluate the fermentation products. 11 LAB strains were selected for further experiments based on the highest products of lactic or acetic acid, including 1 strain of Weissella confusa, 2 of Lactobacillus reuteri and 8 of Lactobacillus plantarum.The API 50 CH fermentation experiment indicated that all of the selected 11 LAB strains utilised most of the carbohydrates. All the strains grew at temperatures between 10 and 45℃ and at a p H of 3.5 to 4.5; however, L. reuteri F7 and F8 tolerated a p H as low as 3.0. All 11 LAB strains showed antibacterial activity against Listeria monocytogens, Escherichia coil, Salmonella sp. and Acetobacter pasteurianus; however, after excluding the effect of organic acids, only F7 and F8 still exhibited antibacterial activity. The present study indicated that the selected 11 LAB strains could be used to prepare silages of RB and MB leaves, especially L. reuteri F7 and F8.

Characteristics of <i>Morus alba</i>L. Cultured by In-Room Hydroponics

Morus alba L., (cv Ichinose) was cultivated by in-room hydroponics. The flavor and texture of leaves were markedly improved enough to be edible. When the contents of 1-deoxynojirimycin (DNJ) and polyphenols were measured in the hydroponic cultivar, DNJ increased in the leaf compared to the field grown M. alba. However, polyphenols, in contrast, decreased compared to the field cultivar. HPLC profiling revealed marked difference in leaf components between hydroponic and field cultivars indicating relative contents of lipophilic polyphenols were increased. The polyphenols contents, especially, lipophilic polyphenols in the root were remarkably high compared to So-Haku-Hi (Sang Bai Pi in Chinese). The anti-obesity effect of the hydroponically grown Morus was further studied in rats by feeding high-fat, high-sucrose (HFHS) diet with and without supplementation of dried leaf and root powders for 15 weeks. As the result, both the leaf and root from the hydroponic cultivar showed potential anti-obesity and anti-hyperlipidemic functions through amelioration of insulin resistance. Differential effects of leaf and root powders indicated that besides DNJ, the lipophilic polyphenols may play a crucial role in the anti-diabetic function of hydroponically grown Morus alba L. The hydroponics will provide an alternate way to cultivate a novel resource of Morus for developing functional foods and medicines.

当年生桑树(Morus alba L.)果枝叶片特征与果实性状的相关性

为揭示叶果间的内在作用规律,以桑树(Morus alba L.)当年生果枝为研究对象,分析不同大小果枝上叶片特征和果实性状的差异及其二者间的相互关系。结果表明:随着果枝由小到大,果枝上的叶片数与总叶干重显著增加,而叶形指数却无显著变化;桑葚数与总桑葚干重也有显著增加,桑葚含水量无显著变化。桑葚数与叶片数、总叶干重、总叶面积及其平均桑葚干重显著正相关。研究结果证实了果实的性状与叶片特征之间存在极密切关系。当年生果枝上的桑葚数量多少取决于果枝上的叶片总数,平均桑葚的干重则与果枝上的平均叶面积显著相关。

桑树(Morus alba L.)原位修复某尾矿区重金属污染土壤

木本植物具有根系发达、生物量大、适应性强等特点,可广泛用于重金属污染土壤修复.本文通过5年的田间修复试验,研究了桑树(Morus alba L.)对污染土壤中重金属的累积和分布特征、土壤中重金属和营养元素有效性含量的变化,来探讨桑树修复某尾矿区污染土壤中Mn、Zn和Cd等重金属的效果.研究结果表明,桑树生物量大,可用于重金属污染土壤的生态修复与景观恢复.田间种植5年后,桑树整株干重每株可达4 kg.桑树对土壤中重金属具有一定的转运和累积能力,地上部分中Cd、Zn和Mn等重金属含量明显大于根部,尤其是叶片中重金属含量明显大于枝和主干中的含量.修复5年后,桑树地上部分Zn和Mn的累积总量可达3277.7 mg·100 m-2和2422.4 mg·100 m-2,且土壤中Mn和Zn含量分别从2192.5 mg·kg-1和103.2 mg·kg-1降低至1790.0 mg·kg-1和85.94 mg·kg-1,同时土壤有效态Mn和Zn分别显著(P<0.05)降低66.0%和28.6%.然而,桑树落叶中Cd、Zn和Mn含量分别可达0.36、64.5、189.2 mg·kg-1.因此,通过定期清除桑树落叶或刈割地上部分,可防止叶片中重金属对土壤造成二次污染,同时削减土壤中重金属含量.同时,经桑树修复5年后土壤中碱解氮、有效磷和速效钾含量均显著(P<0.05)降低,需定期补充相应氮、磷和钾肥来强化桑树修复尾矿区重金属污染土壤.

Correlation between Different Morus alba L. Leaf Nodes and the Change in Leaf Size or Mass

This paper aimed to explore the correlation between Morus alba L. leaf node and its size or mass change and provide a theoretical basis for the development of scientific Morus alba L. cultivation technology. During 2016-2017,the correlation between Morus alba L. leaf node and its size or mass change was investigated and researched. Results showed that through the correlation coefficient analysis,there was a weakly positive correlation between the leaf node and some indicators such as( leaf length ÷ leaf width) and petiole length,there was a weakly negative correlation between the leaf node and some indicators such as leaf width,( leaf length + leaf width),( leaf length × leaf width) and leaf mass,and there was a weakly negative correlation between the leaf node and the other indicators; there was a highly positive correlation between the leaf mass and some indicators such as Morus alba L. leaf length,leaf width,( leaf length + leaf width) and( leaf length × leaf width),there was a significantly positive correlation between the leaf mass and some indicators such as petiole transverse diameter in width and petiole mass,and there was a weakly positive correlation between the leaf mass and some indicators such as petiole transverse diameter in thickness. In terms of significant level of difference,it was found that there was no significant difference in the correlation coefficients between the leaf nodes on the Morus alba L. shoots and some indicators such as leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width) and leaf mass; there was an extremely significant difference in the correlation coefficients between the leaf mass and some indicators such as Morus alba L. leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width),petiole transverse diameter in width and petiole mass; there was no significant difference in the correlation coefficients between the leaf mass and the other indicators. The regression analysis was performed on the leaf node and leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width),( leaf length ÷ leaf width),petiole length,petiole diameter in width,petiole diameter in thickness,petiole mass and leaf mass. Significance F values were0. 4864,0. 1995,0. 2888,0. 3004,0. 2350,0. 2407,0. 8925,0. 5227,0. 7022 and 0. 2000,respectively,that is,there was an extremely significant difference between Morus alba L. leaf node and petiole diameter in width,there was a significant difference between Morus alba L.leaf node and petiole mass,and there was no significant difference between Morus alba L. leaf node and the other indicators. The comparative analysis of quadratic curve regression equation and linear regression equation was performed on some indicators such as leaf length,and it was found that the R value between the leaf node and petiole diameter in width was in line with the logarithmic curve regression equation,and the other R values of quadratic curve regression equation was larger than the other R values of linear regression equation,indicating that different Morus alba L. leaf nodes,leaf length,leaf mass and other indicators were in line with the quadratic curve regression equation. It was concluded that by investigating the leaf node on the Morus alba L. shoots and regression equation,we could predict the leaf size,leaf mass and other indicators,and we could take the leaf node on the Morus alba L. shoots as a main factor affecting the Morus alba L. leaf size and mass.

Plants regenerated from mesophyll protoplasts of white mulberry

Morus alba(white mulberry) mesophyll protoplasts were isolated from leaves of 30-45 day old sterile shoots,with protoplast yields of 2.5 x 107 g-1/F.W. after purification. The protoplasts were cultured in a modified K8P liquid medium containing 0.2 mg/L 2,4-D(2,4- Dichlorophe-noxy acetic acid), 1 mg/L NAA(Naphthyl acetic acid) and 0.5 mg/L BA(6-benzylaminopurine). A low plating density (5 x 104/ml) proved to be favourable to the division of protoplast-derived cells. The first divisioll occurred 4 days after culture, and the division frequency reached 24% at 10 days. A number of cell colonies and microcalli formed in 6 weeks. The microcalli were transferred onto MSB medium with 0.5 mg/L NAA and 0.5 mg/L BA for further proliferation. Shoot formation was initiated when the calli of 3-4 mm in size were transferred onto MSB differentiation medium with 0.1 mg/L NAA and 1 mg/L BA. The frequency of shoot formation was 35%. The shoots of 4-5 cm in height were excised from the callus and rooted on half strength MS medium with 0.5 mg/L IBA and 0.1 mg/L BA. After transplantation into pots, the regenerated plants grew vigorously in the phytotron.

桑树磷脂酶MaPLA1-2D亚型4个基因的克隆与胁迫应答分析

磷脂酶(phospholipase)是一类在植物生长发育和胁迫应答中起重要调控作用的磷脂水解酶,也是一类重要的信号转导酶。而磷脂酶A1(PLA1)在植物应答生物胁迫和非生物胁迫中的功能研究鲜见报道。研究从桑树(Morus alba L.)中克隆了磷脂酶PLA1的1个亚型MaPLA1-2D基因,对其进行了序列分析、组织表达、胁迫诱导表达和蛋白亚细胞定位分析。结果表明,桑树PLA1-2D亚型基因包括4个成员,命名为MaPLA1-2D.1~MaPLA1-2D.4。4个基因在桑树根和叶中高水平表达,蛋白亚细胞定位在叶绿体。序列和进化分析表明MaPLA1-2D基因4个成员与拟南芥AtDAD1基因的保守结构域序列具有较高相似度且进化关系紧密。MaPLA1-2D基因4个成员的启动子含有多种胁迫应答顺式元件和激素响应元件;胁迫诱导表达模式分析表明MaPLA1-2D基因表达受干旱和脱落酸处理显著诱导。以上结果说明,MaPLA1-2D基因与拟南芥DAD同源,可能在桑树非生物胁迫应答中发挥重要功能。

HPLC法同时测定桑白皮中5种成分

目的建立HPLC法同时测定桑白皮Morus alba L.中绿原酸、二氢桑色素、氧化白藜芦醇、桑辛素O、桑根酮C的含有量。方法桑白皮甲醇提取物的分析采用Dimonsiol C18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.2%醋酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长340 nm。结果绿原酸、二氢桑色素、氧化白藜芦醇、桑辛素O、桑根酮C分别在3.25~104.00μg/mL(r=0.999 6)、4.81~154.00μg/mL(r=0.999 7)、5.38~172.00μg/mL(r=0.999 5)、1.88~60.00μg/mL(r=0.999 7)、11.19~358.00μg/mL(r=0.999 5)范围内线性关系良好。平均加样回收率分别为98.98%(RSD=1.70%)、100.73%(RSD=1.35%)、99.71%(RSD=1.64%)、100.07%(RSD=1.11%)、99.90%(RSD=2.16%)。结论该方法准确简便,重复性好,可用于桑白皮药材的质量控制。

不同产地桑椹药材HPLC指纹图谱研究

目的:采用高效液相色谱法建立桑椹药材的HPLC指纹图谱,为其品质控制提供可靠依据。方法:采用HPLC-UV分析桑椹的指纹图谱。色谱柱:Agilent XDB C18柱(4.6 mm×250 mm,5μm),流动相:乙腈-0.3%磷酸溶液梯度洗脱,流速:0.5 mL/min,柱温:35℃,检测波长:290 nm。测定的10批桑椹指纹图谱,应用相似度分析对桑椹药材进行研究。结果:在高效液相指纹图谱中,确定了12个共有峰,经过相似度分析,10批药材中7批相似度较好,其中3批相似度偏低。结论:桑椹药材指纹图谱特征性及专属性强,可为药材的质量控制提供依据。

不同产地桑叶药材UPLC指纹图谱研究

目的建立不同产地桑叶UPLC指纹图谱。方法采用ACQUITY UPLC BEH C18(2.1 mm×50.0 mm,1.7μm)色谱柱,流动相为甲醇-0.2%磷酸水溶液,梯度洗脱,检测波长为358 nm,流速为0.6 ml/min,柱温40℃,进样体积1μl。结果建立了桑叶专属性的UPLC指纹图谱,确定了15个共有峰,不同产地桑叶样品具有较好的相似度。结论 UPLC指纹图谱方法重复性好,简便可靠,可用于桑叶的快速鉴别,可以为桑叶的质量控制和市场应用提供依据。

电导法协同Logistic方程测定不同品种桑树抗寒性

采用电导法研究低温胁迫条件下不同品种桑树枝条相对电导率的变化规律,并利用Logistic方程计算9个桑树品种的半致死温度(LT_(50)),比较其抗寒性。结果表明:9个桑树品种枝条的相对电导率变化曲线均随处理温度的降低呈S形曲线上升,计算枝条的低温半致死温度LT_(50)分别为-29.47℃、-35.42℃、-26.60℃、-32.87℃、-33.63℃、-26.88℃、-29.37℃、-21.03℃和-39.96℃。即9个桑树品种抗寒性由强到弱排序为:向海一号>新桑8号>龙桑一号>冀桑3号>冀桑2号>秋雨>辽鲁11号>桲椤桑>辽育8号。

桑白皮提取物中桑根酮C、桑根酮D及桑辛素的HPLC含量测定

目的:建立HPLC同时测定桑白皮提取物中桑根酮C、桑根酮D及桑辛素的含量测定方法,为其质量控制和评价提供参考。方法:采用InterSustain AQ-C18(250 mm×4.6 mm,5μm)色谱柱;流动相为乙腈(A)-0.1%醋酸水(B),梯度洗脱(0~20 min,45%~70%A;20~26 min,70%~80%A);流速1 mL·min^-1;柱温30℃;检测波长276 nm。结果:3种成分均能实现完全分离,且与其他成分分离度良好。3种成分分别在0.280~4.480μg、0.196~3.136μg、0.054~0.865μg范围内呈良好的线性关系,回归系数分别为1.0000、0.9992、0.9992。桑根酮C、桑根酮D及桑辛素的平均加样回收率(n=6)分别为95.41%、101.02%、101.27%,RSD分别为1.91%、2.31%、1.84%。结论:所用方法操作简便,测定准确,重复性好,可用于桑白皮提取物的质量控制与评价。

桑白皮血清HPLC指纹图谱及入血成分鉴定

目的:建立桑白皮的血清HPLC指纹图谱,并采用UPLC-MS鉴定入血成分,筛选桑白皮有效成分。方法:灌胃给予大鼠桑白皮提取液,取眼眶静脉血,采用溶剂法沉淀蛋白,进样考察,建立桑白皮血清HPLC指纹图谱;采用对照品及UPLC-MS法鉴别桑白皮的入血成分。结果:建立了桑白皮的血清指纹图谱,确定了2个血中移行成分,其中1个为原型成分桑皮苷A。结论:桑皮苷A为桑白皮的入血成分,具有平喘止咳的功效,为桑白皮的有效成分。

α-Glucosidase inhibitors and antioxidants from root bark of Morusalba

Objective: To study the chemical constituents from root bark of Morus alba and their α-glucosidase inhibition and DPPH radical scavenging activities.Methods: The chemical constituents were isolated and purified by repeated column chromatographies on silica gel, Sephadex LH-20, and preparative HPLC. Their structures were elucidated by 1 D and 2 D NMR spectra and HR-ESI-MS.Results: Thirteen compounds 1–13 were isolated and identified. The bioactive assays revealed that compounds 1, 3 and 8 displayed strong α-glucosidase inhibitory activity with IC50 values of(147.1 ± 1.1),(314.1 ± 0.8), and(207.6 ± 0.1) μmol/L, respectively, which were stronger than the positive control of acarbose(418.6 ± 0.1 μmol/L). Compounds 10 and 11 displayed potent DPPH scavenging activity with EC50 values of(2.9 ± 0.1) and(5.0 ± 0.1) μmol/L [EC50 of positive control Vitamin C was(54.8 ± 0.1) μmol/L],respectively.Conclusion: To the best of our knowledge, this is the first report about the compounds 1, 3, and 8 of M.alba with α-glucosidase inhibitory effects.

桑白皮多糖调控circDLGAP4/miR-320对缺氧/复氧诱导心肌细胞损伤的作用

目的研究表明桑白皮多糖具有抗氧化作用,而其对心肌细胞氧化损伤的影响尚不清楚,文章旨在探讨桑白皮多糖对缺氧/复氧诱导的心肌细胞损伤的作用及分子机制。方法H9C2细胞缺氧培养6 h后再复氧6 h,作为缺氧/复氧(H/R)组,不做任何处理的细胞作为正常对照组,用0.1、0.2、0.4μg/mL的桑白皮多糖预处理H9C2细胞,而后进行缺氧/复氧处理,作为H/R+桑白皮多糖低、中、高剂量组;将pcDNA、pcDNA-circDLGAP4转染至H9C2细胞后进行缺氧/复氧处理,记为H/R+pcDNA组、H/R+pcDNA-circDLGAP4组;将si-circDLGAP4转染至H9C2细胞后用0.4μg/mL桑白皮多糖处理,再进行缺氧/复氧处理,记为H/R+桑白皮多糖高剂量+si-circDLGAP4组。细胞计数试剂盒8(CCK-8)、流式细胞术分别检测以上各组细胞活性和凋亡率;丙二醛(MDA)、超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)试剂盒分别检测MDA含量、SOD活性和LDH活性;实时荧光定量PCR(RT-qPCR)检测各组细胞circDLGAP4和miR-320的表达水平。双荧光素酶报告实验检测circDLGAP4和miR-320的靶向关系。结果与正常对照组相比,H/R组H9C2细胞活性降低,凋亡率升高,Bax表达水平升高,Bcl-2表达水平降低(P<0.05);与H/R组相比,H/R+桑白皮多糖低、中、高剂量组H9C2细胞活性升高,凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,呈剂量依赖性(P<0.05)。与正常对照组H9C2细胞MDA[(18.35±1.11)nmL/mL]、LDH(26.81±1.11)U/L,SOD活性[(65.53±2.75)U/L]相比,H/R组H9C2细胞中MDA含量[(95.02±3.67)nmL/mL]及LDH活性[(133.97±5.30)U/L]升高,SOD活性[(11.92±0.63)U/L]降低(P<0.05);与H/R组相比,H/R+桑白皮多糖低、中、高剂量组H9C2细胞中MDA含量[(71.43±3.00)、(53.62±1.69)、(35.81±1.36)nmL/mL]及LDH活性[(106.43±4.00)、(84.50±2.68)、(58.51±1.94)U/L]降低,SOD活性[(24.60±1.14)、(42.37±2.26)、(58.27±2.61)U/L]升高,呈剂量依赖性(P<0.05)。与正常对照组相比,H/R组H9C2细胞中circDLGAP4表达水平降低,miR-320表达水平升高(P<0.05);与H/R组相比,H/R+桑白皮多糖低、中、高剂量组H9C2细胞中circDLGAP4表达水平升高,miR-320表达水平降低,呈剂量依赖性(P<0.05)。与pcDNA组相比,pcDNA-circDLGAP41、2、3组circDLGAP4表达水平升高。与H/R+pcDNA组相比,H/R+pcDNA-circDLGAP4组circDLGAP4表达水平升高,miR-320表达水平降低,MDA含量、LDH活性降低,SOD活性升高;H9C2细胞活性升高,H9C2细胞凋亡率及Bax表达水平降低,而Bcl-2表达水平升高(P<0.05)。与H/R+桑白皮多糖高剂量组相比,H/R+桑白皮多糖高剂量+si-circDLGAP4组circDLGAP4表达水平降低,miR-320表达水平升高,H9C2细胞中MDA含量、LDH活性升高,SOD活性降低,H9C2细胞活性降低,H9C2细胞凋亡率及Bax表达水平升高,而Bcl-2表达水平降低(P<0.05)。circDLGAP4和miR-320的互补序列。wt-circDLGAP4与miR-320共转染的细胞荧光素酶活性较与miR-NC共转染的降低(0.96±0.08 vs 0.35±0.03,P<0.05)。结论桑白皮多糖通过调控circDLGAP4/miR-320对缺氧/复氧诱导的心肌细胞损伤起保护作用。

A New Geranylated 2-Arylbenzofuran from Morus alba

Objective To study the chemical constituents from the root barks of Morus alba. Methods Various chromatographic techniques were used to separate the constituents and their structures were elucidated on the basis of extensive spectroscopic interpretation. Results Six compounds were isolated from the root barks of M. alba. The structure of new compound, named as albafuran D（1）, was established together with known compounds sanggenon H（2）, kuwanon E（3）, kuwanon U（4）, kuwanon F（5）, and benzokuwanon E（6）. Conclusion Compound 1 is a new geranylated 2-arylbenzofuran.