Isolation and Expression Analysis of Two Genes Encoding Cinnamate 4-Hydroxylase from Cotton(Gossypium hirsutum)

Two genes （GhC4H1 and GhC4H2） that encode putative cotton cinnamate 4-hydroxylases that catalyze the second step in the phenylpropanoid pathway were isolated from developing cotton fibers. GhC4H1 and GhC4H2 each contain open reading frames of 1 518 base pairs （bp） in length and both encode proteins consisting of 505 amino acid residues. They are 90.89% identical to each other at the amino acid sequence level and belong to class I of plant C4Hs. GhC4H1 and GhC4H2 genomic DNA are 2 247 and 2 161 bp long, respectively, and contain two introns located at conserved positions relative to the coding sequence. GhC4HI and GhC4H2 promoters were isolated and found to contain many cis-elements （boxes P, L and AC-1 element） previously identified in the promoters of other phenylpropanoid pathway genes. Histochemical staining showed GUS expression driven by the GhC4H1 and GhC4H2 promoters in ovules and fibers tissues. GhC4H1 and GhC4H2 were also widely expressed in other cotton tissues. GhC4H2 expression reached its highest level during the elongation stage of fiber development, whereas GhC4H1 expression increased during the secondary wall development period in cotton fibers. Our results contribute to a better understanding of the biochemical role of GhC4H1 and GhC4H2 in cotton fiber development.

Cinnamic acid regulates the intestinal microbiome and short-chain fatty acids to treat slow transit constipation

BACKGROUND Slow transit constipation(STC)is a disorder with delayed colonic transit.Cinnamic acid(CA)is an organic acid in natural plants,such as Radix Scrophulariae(Xuan Shen),with low toxicity and biological activities to modulate the intestinal microbiome.AIM To explore the potential effects of CA on the intestinal microbiome and the primary endogenous metabolites-short-chain fatty acids(SCFAs)and evaluate the therapeutic effects of CA in STC.METHODS Loperamide was applied to induce STC in mice.The treatment effects of CA on STC mice were assessed from the 24 h defecations,fecal moisture and intestinal transit rate.The enteric neurotransmitters:5-hydroxytryptamine(5-HT)and vasoactive intestinal peptide(VIP)were determined by the enzyme-linked immunosorbent assay.Hematoxylin-eosin and Alcian blue and Periodic acid Schiff staining were used to evaluate intestinal mucosa's histopathological performance and secretory function.16S rDNA was employed to analyze the composition and abundance of the intestinal microbiome.The SCFAs in stool samples were quantitatively detected by gas chromatography-mass spectrometry.RESULTS CA ameliorated the symptoms of STC and treated STC effectively.CA ameliorated the infiltration of neutrophils and lymphocytes,increased the number of goblet cells and acidic mucus secretion of the mucosa.In addition,CA significantly increased the concentration of 5-HT and reduced VIP.CA significantly improved the diversity and abundance of the beneficial microbiome.Furthermore,the production of SCFAs[including acetic acid(AA),butyric acid(BA),propionic acid(PA)and valeric acid(VA)]was significantly promoted by CA.The changed abundance of Firmicutes,Akkermansia,Lachnoclostridium,Monoglobus,UCG.005,Paenalcaligenes,Psychrobacter and Acinetobacter were involved in the production of AA,BA,PA and VA.CONCLUSION CA could treat STC effectively by ameliorating the composition and abundance of the intestinal microbiome to regulate the production of SCFAs.

New Process for the Synthesis of A Sun-screening Agent Isooctyl P-methoxy Cinnamate

A sun-screening agent isooctyl p-methoxy cinnamate was prepared by reaction of p-methoxyl benzaldehyde with isooctyl acetate using titanium tetrachloride as catalyst. The optimum conditions are as follows: molar ratio of p-methoxyl benzaldehyde to isooctyl acetate and titanium tetrachloride is 1 : 1.1 : 0.5; reaction temperature is 30°C; and reaction time is 6 h. The yield of isooctyl p-methoxy cinnamate can reach 98%. The method of the synthesis of isooctyl p-methoxy cinnamate is efficient and economical.

N-(苯乙基)肉桂酰胺类化合物的合成及其抗氧化活性研究

植物N-(苯乙基)肉桂酰胺类化合物是一类重要的酰胺类活性成分,为寻求更高药理活性的肉桂酰胺类先导化合物,以苯甲醛、丙二酸为原料,设计合成了9种N-(苯乙基)肉桂酰胺类化合物,结构经过^(1)H NMR、^(13)C NMR、IR和MS鉴定。以抗坏血酸为对照,测定9种化合物清除·OH、DPPH·及ABTS+·的效果。抗氧化实验结果表明,部分化合物对·OH的清除效果优于抗坏血酸;9种化合物具有一定清除DPPH·及ABTS+·的能力,但效果不如抗坏血酸。该类化合物可进一步结构优化,探究其抗阿尔茨海默病、抗肿瘤等活性研究,也可作为抗氧化剂应用于食品、药品领域。

肉桂酸和邻苯二甲酸对沙芥属植物幼苗光合特性的影响

【目的】探讨酚酸类自毒物质对沙芥属植物生长及光合特性的影响,为沙芥属植物的野生资源保护和开发利用提供理论依据。【方法】以沙芥和斧翅沙芥幼苗为试验材料,选择肉桂酸和邻苯二甲酸2种自毒物质,采用盆栽试验,设置0,0.01,0.1,1,10 mmol/L浓度梯度处理,考察幼苗生长、叶绿素含量、叶绿素荧光参数和气体交换参数的变化特征。【结果】(1)肉桂酸、邻苯二甲酸对沙芥幼苗生长均表现为低浓度促进高浓度抑制,而对斧翅沙芥生长则表现为抑制作用,且均在浓度为10 mmol/L时对生长抑制作用最显著。(2)在不同浓度肉桂酸和邻苯二甲酸处理后,沙芥和斧翅沙芥幼苗叶片的叶绿素a、叶绿素b、叶绿素总量均出现不同程度下降。(3)沙芥和斧翅沙芥叶片净光合速率、蒸腾速率、气孔导度在肉桂酸和邻苯二甲酸浓度为10 mmol/L显著低于对照,胞间CO_(2)浓度此时无显著变化,其光合速率降低主要由非气孔因素所致。(4)沙芥和斧翅沙芥的最大光化学效率(F_(v)/F_(m))、实际光化学效率(Ф_(PSⅡ))和光化学淬灭系数(qP)均在肉桂酸和邻苯二甲酸浓度为10 mmol/L显著低于对照,而此时非光化学淬灭系数(NPQ)则显著高于对照。【结论】高浓度邻苯二甲酸和肉桂酸抑制沙芥和斧翅沙芥幼苗的光合作用,降低其光合速率,导致它们叶片的PSⅡ反应中心活性和开放程度受损,进而影响其正常生长;肉桂酸和邻苯二甲酸对沙芥属植物光合特性的影响存在差异,斧翅沙芥幼苗受到影响更大。

基于网络药理学与实验验证探讨肉桂酸治疗慢性心力衰竭的作用机制

目的采用网络药理学、分子对接及体外实验验证的方法,探讨肉桂酸治疗慢性心力衰竭(chronic heart failure,CHF)的潜在靶点及作用机制。方法通过网络药理学在线数据库预测肉桂酸靶点及CHF靶点,取交集靶点于String数据库构建蛋白互作网络,通过Cytoscape 3.7.2软件进行可视化分析,筛选出核心靶点,使用Metascape数据库进行GO和KEGG富集分析,预测肉桂酸治疗CHF的作用机制,采用分子对接技术及体外实验验证上述结果。结果网络药理学分析结果发现,肉桂酸潜在靶点187个,与6094个CHF疾病靶点相交,得到132个交集靶点,涉及CASP3、JUN、PTGS2、MMP9、TLR4等10个核心靶点。GO和KEGG富集分析显示,肉桂酸治疗CHF的作用主要与细胞对活性氧的反应、对氧化应激的反应等生物过程有关,涉及细胞凋亡等信号通路。分子对接结果显示,肉桂酸与10个核心靶点均具有良好的结合活性。体外实验结果显示,肉桂酸能显著降低心肌细胞H9c2凋亡率及活性氧水平(P<0.05),降低c-Jun、PTGS2、MMP9、TLR4 mRNA表达水平(P<0.05)和caspase-9、caspase-3、Bax蛋白表达水平(P<0.05),升高Bcl-2蛋白表达水平(P<0.05);使用乙酰半胱氨酸后,细胞凋亡率、活性氧水平及Bax蛋白表达水平显著降低(P<0.05),Bcl-2蛋白表达水平显著升高(P<0.05),与肉桂酸作用一致。结论肉桂酸能够调节多个信号靶点、抑制心肌细胞氧化应激损伤及细胞凋亡,发挥治疗CHF的作用,肉桂酸是苓桂术甘汤防治CHF的重要物质基础之一。

Anticancer Activities of Substituted Cinnamic Acid Phenethyl Esters on Human Cancer Cell Lines

Caffeic acid phenethyl ester (CAPE) and sixteen substituted cinnamic acid phenethyl esters were prepared via conventional procedures in order to test their in vitro anticancer activities by either MTT assay or SRB assay on six different human cancer cell lines. The results indicated that in the concentration of 10μmol·L^-1 the lead compmuM CAPE possessed anficancer activities against human HL-60, Bel-7402, and Hela cell lines, and two other compounds possessed potent anticancer activities against Bel-7402 and Hela cell lines.

三种药剂对水绵控制效果的比较分析

选择硫酸铜、扑草净以及肉桂酸为控藻药剂,以常见的丝状藻-水绵为受试对象,比较了三种药剂的控藻效果。研究结果表明,三种药剂的施用对水绵生长、代谢及抗逆功能均造成了影响,但影响程度因药剂类型、药剂浓度及施用时间而异。相同浓度条件下,硫酸铜、扑草净的控藻效果均优于肉桂酸,但硫酸铜的施用会造成环境累积和生物放大等危害。结合控藻有效性、安全性和经济性,选择1 mg/L扑草净为最优方案。

富马酸与肉桂醛联用调节产肠毒素型大肠杆菌诱导猪肠上皮细胞氧化应激的分子机制

本试验旨在研究富马酸(FA)与肉桂醛(CA)联用调节产肠毒素型大肠杆菌(ETEC)K88诱导猪肠上皮细胞IPEC⁃J2氧化应激的分子机制。试验选用IPEC⁃J2细胞建立氧化损伤模型,用不同浓度FA和CA处理IPEC⁃J2细胞12和24 h,并用CCK⁃8法检测细胞活力,确定最佳处理浓度和时间;以最佳处理浓度的FA和CA预处理细胞,ETEC K88感染细胞3、6、12和24 h,检测其活菌黏附率,采用酶联免疫吸附测定(ELISA)检测细胞因子含量和抗氧化指标,并用实时荧光定量PCR(RT⁃qPCR)测定热休克蛋白70(Hsp70)和核因子-κB(NF⁃κB)信号通路相关基因mRNA相对表达量。结果表明:1)FA和CA的最佳处理浓度分别为1.00 mg/mL和1.00μL/mL,最适培养时间为12 h。2)添加FA和CA可有效抑制ETEC K88黏附IPEC⁃J2细胞,当ETEC K88感染细胞3 h时其黏附率显著下降(P<0.05),6、12和24 h时极显著下降(P<0.01)。3)添加FA和CA可极显著降低ETEC K88感染细胞中促炎性细胞因子白细胞介素-8(IL⁃8)和肿瘤坏死因子-α(TNF⁃α)含量(P<0.01),极显著提高抗炎性细胞因子白细胞介素-10(IL⁃10)和转化生长因子-β(TGF⁃β)含量(P<0.01)。4)ETEC K88通过激活NF⁃κB信号通路诱导IPEC⁃J2细胞氧化损伤,添加FA和CA可上调Hsp70 mRNA相对表达量并抑制NF⁃κB信号通路缓解IPEC⁃J2细胞氧化应激。5)添加FA和CA可显著提高ETEC K88感染细胞中超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH⁃Px)活性(P<0.05),极显著降低丙二醛(MDA)含量(P<0.01)。综上所述,FA与CA联用可调节炎性细胞因子和氧化应激蛋白平衡并抑制ETEC K88黏附IPEC⁃J2细胞,其可能是通过上调Hsp70抑制NF⁃κB信号通路,增强IPEC⁃J2细胞的抗氧化和抗炎能力,从而缓解ETEC K88诱导的IPEC⁃J2细胞氧化应激。

分散液液微萃取-高效液相测定食用植物油中对羟基肉桂酸和橙皮素含量

通过分散液液微萃取-高效液相(DLLME-HPLC)建立一种同时测定食用植物油中对羟基肉桂酸和橙皮素的分析方法。以正己烷为分散剂,以含体积分数5%氨水(500μL)和体积分数10%乙腈的水溶液为萃取剂萃取样品,以体积分数0.3%三氟乙酸溶液-乙腈为流动相,在柱温25℃条件下进行梯度洗脱,萃取液放置过夜后再进行测定。结果表明:样品峰面积的相对标准偏差均小于3%,对羟基肉桂酸和橙皮素在各自质量浓度范围内的线性相关系数均大于0.99,检测限分别为8.7、6.2 ng/g,定量限分别为26.1、18.5 ng/g,回收率分别为93.1%~103.7%、100.5%~111.1%。

Aldol缩合-水解“一锅”法制备反式肉桂酸——推荐一个改进的有机合成实验

肉桂酸是一种重要的有机合成中间体以及精细化工原料,广泛应用于多个领域。肉桂酸制备是本科生有机化学实验教学中的重要内容之一,常按照经典的以苯甲醛和乙酸酐为原料、醋酸钾/碳酸钾为催化剂的Perkin反应来进行。本文提出了一种新的方法——Aldol缩合-水解法,用于制备反式肉桂酸。该法以苯甲醛、乙酸乙酯和叔丁醇钾为原料进行反应,再以“一锅”的方式依次进行蒸馏、加碱水解、蒸馏和酸化即得反式肉桂酸纯品,具有原料/试剂廉价易得、用量少、实验装置简单、能耗小、操作便捷、产率高、耗时较短和环境污染小等优点,非常值得推荐用于本科生实验教学,更有利于对学生在掌握多方面基础理论知识和实验技能的同时,拓展便捷高效合成有机物的创新思维和提高节能环保意识等综合素质的培养。

GC-MS/MS法同时测定十滴水中4种挥发性成分的含量

目的 建立同时测定十滴水中樟脑、桉油精、桂皮醛、反式茴香脑等4种挥发性成分质量浓度的GC-MS/M法。方法 色谱柱采用Agilent HP-5MS UI柱(30 m×0.25 mm, 0.25μm);程序升温:初始温度65℃,保持2 min,以每分钟6℃的速率升至150℃,保持10 min;采用标准电子电离(EI),多反应监测(MRM)模式进行检测。结果 桉油精、樟脑、桂皮醛、反式茴香脑分别在1.03~51.34、5.85~292.26、0.98~48.86、0.99~49.85μg/mL范围内与峰面积线性关系良好,相关系数r分别为0.999 2、0.999 8、0.999 8、0.999 9;平均加样回收率分别为97.76%、96.94%、106.21%、102.50%,RSD分别为2.86%、2.54%、2.04%、2.92%;精密度、重复性试验RSD值在1.27%~1.90%之间。结论 该方法操作简便,结果准确且重现性好,可为十滴水的质量控制和质量评价提供依据。

HPLC法测定金丹附延颗粒中肉桂酸和桂皮醛的含量

目的:建立金丹附延颗粒中肉桂酸和桂皮醛的HPLC含量测定方法。方法:采用C 18色谱柱,流动相为乙腈和0.1%磷酸溶液,检测器检测波长设置为290 nm,输液泵流速1.0 mL·min^(-1),色谱柱柱温设置为25℃。结果:肉桂酸、桂皮醛分别在0.2079~10.3938μg·mL^(-1)、0.2025~10.1270μg·mL^(-1)范围内线性关系良好(r=1.0000),回收率分别为100.3%、102.6%,回收率RSD分别为1.57%、1.82%。结论:该方法简单、稳定,可用于金丹附延颗粒中肉桂酸和桂皮醛的含量测定。

本科教学实验肉桂酸的制备及定性、定量分析

以苯甲醛和醋酸酐为原料,无水碳酸钾或醋酸钾作为碱性催化剂,经Perkin反应制备肉桂酸。通过熔点、红外光谱、核磁共振氢谱、碳谱对所得产物进行定性分析,并用核磁共振氢谱完成定量分析。结果表明碳酸钾法在核磁收率上优于醋酸钾法。

亲水性超级交联肉桂酸的合成及其对印染废水的吸附研究

为了获得亲水性的超交联聚合物印染废水吸附剂,并探究其吸附效果,在80℃下,通过傅克烷基化反应,将含有羧基的肉桂酸单体连接成为亲水性超级交联多孔吸附剂,考察了超级交联肉桂酸吸附剂对亚甲基蓝的吸附性能,探究了其吸附规律。结果显示,在室温下(23℃),当pH值为5.8时,超级交联肉桂酸多孔吸附剂对175 mg/L亚甲基蓝溶液的吸附容量为347.78 mg/g。当吸附剂投加量为0.02 g且在pH值为10时吸附容量达到较高水平的840.55 mg/g。吸附动力学和等温线研究结果表明吸附过程更符合准二阶动力学和Langmuir方程。超级交联肉桂酸具有高吸附容量及良好的亲水性,是一种优异的印染废水吸附剂。

咽炎片中5个成分的含量测定及质量分析

目的建立HPLC法同时测定咽炎片中5个成分芍药苷、黄芩苷、哈巴俄苷、肉桂酸、款冬酮的含量测定方法,考察咽炎片的整体质量情况。方法采用Ag ilent ZORBAX SB-C 18色谱柱(4.6 mm×250 mm,5μm);以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为230 nm(芍药苷)、280 nm(黄芩苷、哈巴俄苷、肉桂酸)、213 nm(款冬酮);柱温为30℃;流速为0.8 mL·min^(-1)。采用SPSS 20.0软件对5个成分的含量测定结果进行统计学分析。结果芍药苷、黄芩苷、哈巴俄苷、肉桂酸、款冬酮分别在6.35~402.74μg·mL^(-1)、1.89~120.91μg·mL^(-1)、1.22~77.95μg·mL^(-1)、1.04~66.39μg·mL^(-1)、1.25~80.40μg·mL^(-1)范围内线性关系良好,平均加样回收率(n=9)在96.61%~99.91%。通过测定50批咽炎片中5个成分的含量,发现咽炎片的质量情况一般,部分生产企业存在原料药把关不严、使用劣质药材投料或未按处方足量投料的问题,现行咽炎片质量标准有待完善。结论该方法精密度、稳定性、重复性良好,为咽炎片的质量控制提供科学依据。

肉桂酸衍生物的太赫兹光谱及弱相互作用分析

利用太赫兹时域光谱技术(THz-TDS)测试了对羟基肉桂酸(PCA)、反式-2-羟基肉桂酸(OCA)和4-氟肉桂酸(4-FCA)三种肉桂酸的衍生物(CADs)在0.5~3.5 THz范围内的吸收峰。基于密度泛函理论(DFT),借助振动模式自动关联判定方法(VMARD)对三种样品THz吸收峰的来源进行了指认。最后采用分子力场能量分解法(EDAFF)分析了分子体系的弱相互作用力形式,通过VMD绘制原子着色图进行了可视化分析,并研究了其原子在分子体系中对弱相互作用力的贡献类型和强弱。研究结果表明,基于THz-TDS、DFT与VMARD、EDA-FF方法结合不但能有效地辨别同分异构体及结构近似的有机分子,而且也能为其生化功能揭示提供有价值的参考数据。

Effects of Cinnamic Acid on Bacterial Community Diversity in Rhizosphere Soil of Cucumber Seedlings Under Salt Stress

To investigate the effects of a plant autotoxin, cinnamic acid, on bacterial communities in the rhizosphere soil of cucumber seedlings under salt stress, we used cucumber as the experimental material, cinnamic acid as the autotoxin, and NaCl to apply salt stress. Bacterial communities in the rhizosphere soil were analyzed using polymerase chain reaction （PCR）, denaturing gradient gel electrophoresis （DGGE）, and clone sequencing. Salt stress decreased the diversity of bacterial species in rhizosphere soil of cucumber seedlings at several growth stages. Cinnamic acid exacerbated the effects of salt stress at high concentrations, but alleviated its effects at low concentrations. Cloning and sequencing results indicated that DGGE bands amplified from soil samples showed high homology to uncultured bacterial species. Cinnamic acid at 50 mg kg^-1 soil improved cucumber growth and was the most effective treatment to alleviate the effects of salt stress on bacterial communities.

Synthesis and Crystal Structure of (E)-4-(Benzyloxy)-2-(cinnamoyloxy)-N,N,N-trimethyl-4-oxobutan-1-aminium Chloride as a Double-prodrug

A novel hydrochloride quaternary ammonium salt （E）-4-（benzyloxy）-2-（cinnamo- yloxy）-N,N,N-trimethyl-4-oxobutan-l-aminium chloride （Cz3HzgNO4Cl2, Mr = 454.37） has been synthesized via the sequence of acetylation and esterification by using L-carnitine （L-4-N-trimethy- lammonium-3-hydroxybutyric acid, LC） and cinnamic acid as the starting materials, and its crystal structure was determined by single-crystal X-ray diffraction method. The crystal belongs to monoclinic, space group P212121 with a = 10.1670（4）, b = 10.4488（4）, c = 22.9795（11）A, V = 2441.18（18）A3, Z = 4, Dc = 1.236 g/cm3,μ（MoKa） = 0.293 mm-1, F（000） = 960, Flack factor = -0.01（11）, the final R = 0.0489 and wR = 0.1550 for 3350 observed reflections （I 〉 2σ（/）） and R = 0.0953 for all 5648 unique reflections. The crystal structure involves a conjugated system which shows a reverse olefin structure.

RP-HPLC Determination and Pharmacokinetic Comparison of Cinnamic Acid in Rat Plasma After Administration of Di-Gu-Pi Decoction and Pure Cinnamic Acid

A sensitive, simple, and accurate method was developed for the determination and pharmacokinetic comparison of cinnamic acid in rat plasma after the administration of a Traditional Chinese Medicinal preparation, Di-Gu-Pi decoction, and pure cinnamic acid using RP-HPLC. Di-Gu-Pi was extracted with 5% aqueous sodium bicarbonate, which was followed by purification with ion exchange column chromatography. The plasma samples taken from rats were deproteinized with methanol. The reversed-phase （HPLC） system with a Diamonsil C18 column and methanol-acetonitfile-water （8： 32： 60, volume ratio） （adjusted to pH = 3.0 with glacial acetic acid） as the mobile phase was employed for the separation of cinnamic acid in the plasma samples. The detection was set at 272 nm and 3-（p-fluoro-phenyl）-2-propenoic acid was chosen as the internal standard. The calibration curve was linear in a range from 0. 10 to 25.0μg/mL （R2 = 0. 9988, n = 9）. The precision was 3.42%-10. 10%; the between-day precision was 2. 84%-8.91% ; the accuracy was - 1.51%-1.26% ; the mean recovery was 99. 9%. The method was found to be sensitive, simple, accurate and appropriate for the determination of cinnamic acid.