Antidiabetic activity of Callicarpa nudiflora extract in type 2 diabetic rats via activation of the AMPK-ACC pathway

Objective:To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotoc-ininduced diabetic rats.Methods:The chemical constituents in Callicarpa nudiflora extract were identified by UPLCQ-TOF-MS.Callicarpa nudiflora extract(0.15 and 0.3 g/kg)was orally administered to streptozotocin-induced diabetic rats for 42 d.The effects of Callicarpa nudiflora extract on body weight,blood glucose,serum insulin,total cholesterol,triglyceride,LDL-C and HDL-C were investigated,and its effect on liver and pancreatic pathology was assessed by histopathological analysis.Moreover,the expression levels of adenosine 5’-monophosphate-activated protein kinase(AMPK),glucose transporter type 4(GLUT4),phospho-AMPK/AMPK,and p-acetylcoA carboxylase(P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction,Western blotting,and immunohistochemistry.Results:A total of 34 compounds,including 8 iridoids,14 phenylpropanoids,and 12 flavonoids,were identified from Callicarpa nudiflora extract.Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels,including serum insulin,total cholesterol,triglyceride,LDL-C,and HDL-C.Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes.Furthermore,Callicarpa nudiflora extract increased the expression levels of phospho-AMPK,GLUT4,P-ACC,and insulin receptor substrate-1 and decreased the expression level of PPAR r in diabetic rats.Conclusions:Callicarpa nudiflora extract improved oral glucose tolerance,lipid metabolism,insulin resistance,and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.

Study on Coagulation Activity of Callicarpa nudiflora

[Objective] This study aimed to study the coagulation effect of Callicarpa nudiflora. [Method] The effects of CaUtcarpa nudiflora on blood coagulation system of mice were investigated by measuring prothmmbin time （PT）, thrombin time（TT）, activated partial thromboplastin time （APTT） and plasma fibrinogen （FIB）, [Result] Compared with the control group, the crude extracts and ：eluted parts of Ca#/carpa nudiflora all significantly shortened PT, APTT and TT, and increased the content of FIB in mice, [Coadadon] The crude extracts and eluted parts of Callicarpa nudiflora have a coagulation-promoting effect through affecting several links of the coagulation process in mice,

Promoting wound healing effect of the extract of callicarpa nudiflora and preparation technology of the external ointment

Objective: To study the effect of the extracts from Callicarpa nudiflora on wound healing and the preparation of ointment. Methods: Totally 50 KM mice were randomly divided into five groups, which were negative control group ,positive control group, high, middle and low dose groups of Callicarpa nudiflora ointment. Mice ear swelling test was conducted to investigate the effect of Callicarpa nudiflora induced by xylene, Bacteriostasis experiments were conducted to observe the bacteriostasis effects of Callicarpa nudiflora on five indicator bacteria, i.e. Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, and Staphylococcus aureus in vitro. A total of 30 new zealand white rabbit with half male and half female were randomly divided into five groups, which were negative control group, positive control group, high, middle and low groups of Callicarpa nudiflora ointment. The rabbits were treated with sterilizing blade and treated with external medicine to observe the effect of the extracts from Callicarpa nudiflora on wound healing. Orthogonal test was used to determine the preparation method of Callicarpa nudiflora ointment,Physical properties, centrifugal stratification test, heat and cold resistance test were conducted. Results:Bacteriostasis experiments in vitro showed high dose group of Callicarpa nudiflora extracts got 16.8 cm antimicrobial zone of Pseudomonas aeruginosa. The experimental results showed that the ear swelling degree of mice induced by xylene was significantly reduced in the middle and high dose groups. All the treatment groups could promote skin healing of rabbit significantly compared with negative control group. There were significantly differences between all the treatment groups and negative control group on the wound healing rate. The optimum preparation process was determined by orthogonal screening test. The optimal formula was as follows: Twain-801.2 g, glycerin monostearate 4.7 g, glycerinum 0.9 g, liquid paraffin 10.8 g, Span801.4 g,solid paraffin 1.4 g,vaseline 0.9 g. The emulsification temperature was 65 ℃,and emulsification time was 25 min. Conclusions: The extract of Callicarpa nudiflora can be used as external preparation and the preparation is in compliance with Chinese pharmacopoeia 2015.

Neo-lignans in the Seed Crusts of Trewia nudiflora

Four new neo-lignans were isolated from the chloroform portion of 70% ethanol extract of the seed crusts of Trewia nudiflora L., and were determined to be 9'-methoxy-7'-en-3',8:4',7-di-epoxyneolignan-3,4,9-triol l, 9'-methoxy-7'-en-3',7:4',8-diepoxyneolignan-3,4,9-triol 2, 9'- ethoxy-7'-en-3',8:4',7-diepoxyneolignan-3,4,9-triol 3, 9'-butoxy-7'-en-3',8:4',7-diepoxy-neolignan -3,4,9-triol 4, respectively, on the basis of spectral data. The antimicrobial bioassays showed that 4 had evident inhibitory activity against Mycobacterium tuberculosis.

Quality standard study of Li folk medicine callicarpa nudiflora powder

Objective:To establish the quality standard of Li folk medicine callicarpa nudiflora powder by modern analytical method.Methods:TLC was used to identify the powder.Five heavy metals and harmful elements(Pb,Cd,As,Hg and Cu)in the powder were determined by flame atomic absorption spectrometry and hydride atomic fluorescence spectrometry.The content of luteolin and Mullein glycoside in the powder was determined by HPLC.Results:the fluorescent spots of TLC identification of powder were clear,and the separation of the two main components was good.It has good specificity,reproducibility and durability,which can be used for the identification of the powder.The inspection items of heavy metals for the identification of powder were established.It was stipulated that:according to the determination method,the lead should not exceed 5 mg/kg;the cadmium should not exceed 0.3 mg/kg;the arsenic should not exceed 2 mg/kg;the mercury should not exceed 0.2 mg/kg;the copper should not exceed 20 mg/kg.The content of luteolin and Mullein glycoside in the powder were determined by HPLC and the content inspection item of the powder was established.It was stipulated that the content of luteolin and Mullein glycoside in the powder should not be less than 0.48%and 5.83%,which can be used for the content limit inspection of the powder.Conclusion:the quality standard method established by our institute is advanced and operable,and can be used for the quality control of the powder.

Study on the preparation and quality standard of Callicarpa nudiflora film

Objective:To screen the best prescription and technology of Callicarpa nudiflora film,and to study and establish the quality standard of the flim.Methods:The orthogonal test was used to investigate the effect of PVA content,glycerin content,ethanol and water ratio on the comprehensive score of the coating agent;the general inspection method and content determination method were used to control the quality of Callicarpa nudiflora film,and the quality standard of the film was established.Results:The best formulation was selected as polyvinyl alcohol 6.0g,glycerin 2ml,ethanol:water 1:1.The appearance,stability,heat and cold resistance of the film were good,and the quality standard of the film was established.Conclusion:The formulation is simple and feasible,and the quality control method of the film is stable and feasible.

基于UPLC-Q-Exactive-Orbitrap-MS、网络药理学和实验验证探讨裸花紫珠抗H1N1活性成分及其作用机制

本实验首先对裸花紫珠的提取物进行萃取,得到不同的萃取部位,然后对各部位进行抗甲型流感病毒H1N1型(H1N1)活性筛选,确定裸花紫珠乙酸乙酯萃取部位(EACN)具有较好的体外抗H1N1的活性。采用超高效液相色谱-四级杆-静电场轨道阱联用(UPLC-Q-Exactive-Orbitrap-MS)技术对EACN进行定性分析。基于定性分析的结果,进一步使用数据挖掘和网络药理学,评估药物的活性成分、关键靶点和关键通路,最后使用H1N1感染BALB/c小鼠模型对预测结果进行验证。结果显示,在各个萃取部位中,乙酸乙酯部位体外抗H1N1效果最佳,半数抑制浓度(IC_(50))为51μg/mL,半数细胞毒浓度(CC_(50))为859.4μg/mL,选择指数(SI,CC_(50)/IC_(50))为16.85。从EACN中鉴定出34种化合物,在此基础上,利用网络药理学,筛选出和EACN抗H1N1相关的药物靶点67个,靶点共涉及生物过程491个,与抗H1N1相关的通路147个。H1N1感染BALB/c小鼠实验结果发现,EACN能改善H1N1感染引起的体重降低和肺指数的增加,降低小鼠肺组织的病毒载量,减轻小鼠肺组织的病理损伤,降低外周血血清中TNF-α、IFN-γ、IL-1β水平。实验结果提示EACN可能从直接抑制病毒、抗炎作用和免疫调节作用等多个途径发挥抗H1N1的作用。

南药裸花紫珠的药理活性及其在单胃动物生产中的应用

裸花紫珠可作为新型绿色植物饲料添加剂应用于动物生产中。裸花紫珠的主要化学成分包括黄酮类、苯丙素类、萜类、挥发油类等,这些成分在抗炎、止血、抗菌和增强免疫等方面发挥着不同的药理作用,可在动物生产中应用。为了推动裸花紫珠进一步的开发利用,文章总结了裸花紫珠药理活性和作用机制,综述其在单胃动物生产研究中的进展,为裸花紫珠后续的开发提供参考。

添加裸花紫珠对王草青贮发酵品质和体外产气特性的影响

本试验旨在研究裸花紫珠(Callicarpa Nudiflora Hook.et Arn.)不同添加形式和添加量对王草(Pennisetum purpureum×P.americanum,17.38%DM)青贮发酵品质、营养成分和体外产气特性的影响。试验以王草为原料,设置对照(CK),按王草200 g鲜重比例添加裸花紫珠水提物(ST)1%,2.5%,5%,鲜样混贮(XZ)2%,4%,6%,8%,裸花紫珠粉末(LF)0.5%,1.5%,2.5%,5%,青贮30天后,取样进行分析。结果表明:各处理与CK比较,pH值均显著降低(P<0.05),8%XZ和5%LF的乳酸含量显著高于其它处理组(P<0.01)。5%LF的干物质含量(DM)均显著高于其它处理组,中性洗涤纤维和酸性洗涤纤维含量均显著低于其它处理组(P<0.05)。8%XZ和5%LF的体外干物质消化率最高(P<0.01)。5%LF的产气量显著高于其它处理组(P<0.01)。综上,添加裸花紫珠鲜样和粉末均能改善王草青贮饲料营养价值、发酵品质和体外产气特性,其中添加5%裸花紫珠粉末效果最佳。

裸花紫珠粉对肉鸡生长性能及肠道健康的影响

为探究不同剂量裸花紫珠粉(Callicarpa nudiflora Hook,CH)对肉鸡生长性能、肠道抗氧化能力和肠道屏障功能的影响,试验选取1日龄健康海南文昌鸡240只,随机分为4个处理组,每个处理组4个重复,每个重复15只鸡。空白组(CON组)饲喂基础饲粮,试验组分别在基础饲粮中添加裸花紫珠粉1 g/kg(CH-1组)、3 g/kg(CH-3组)和5 mg/kg(CH-5组),自由采食饮水,试验期28 d。结果显示:(1)CH-3组的日采食量和日增重较CON组增加了7.2%和11.9%(P<0.05)。(2)CH-1组肉鸡法氏囊指数和血清免疫球蛋白M(IgM)含量较CON组提高了48.8%和16.6%(P<0.05);CH-1组和CH-3组肉鸡血清免疫球蛋白G(IgG)含量较CON组提高了11.2%和12.4%(P<0.05)。(3)CH-1组肉鸡丙二醛(MDA)含量较CON组降低了22.4%(P<0.05);CH-1组肉鸡血清超氧化物歧化酶(SOD)活性提高了12.7%(P<0.05)。(4)CH-1组肉鸡回肠的绒毛高度(VH)和绒隐比(V/C)较CON组提高了19.1%和18.6%(P<0.05)。(5)CH-1组肉鸡回肠葡萄糖转运蛋白-2(GLUT-2)、谷胱甘肽过氧化物酶(GSH-Px)、带状闭合蛋白-1(ZO-1)、咬合蛋白(Occludin)、白介素-4(IL-4)和白介素-10(IL-10)基因表达量较CON组提高了41.0%、32.4%、39.6%、38.5%、23.8%和38.0%(P<0.05);CH-3组肉鸡回肠屏障基因闭合蛋白(Claudin)基因表达量较CON组提高了62.4%,CH-3组肉鸡回肠髓样分化因子基因(MyD88)基因表达量较CON组降低了14.5%,CH-5组肉鸡回肠超氧化物歧化酶-1(SOD1)和阳离子氨基酸转运蛋白(CAT)基因表达量较CON组提高了39.4%和48.5%(P<0.05);各剂量组肉鸡回肠Nrf2基因表达量较CON组分别提高了39.4%、30.3%、41.3%(P<0.05)。综上所述,日粮添加1 g/kg裸花紫珠粉可提高肉鸡生长性能,增强机体抗氧化功能和免疫力,促进肠道绒毛发育。

3, 4-seco-Labdane diterpenoids from the leaves of Callicarpa nudiflora with anti-inflammatory effects

Four new 3,4-seco-labdane diterpenoids,nudiflopenes J-M,were isolated from the leaves of Callicarpa nudiflora along with six known compounds.The structures of these diterpenoids were determined by comprehensive spectroscopic analysis.All the isolated compounds were evaluated for their inhibitory effects on NO production in LPS-stimulated RPMs and RAW264.7 cells.The results suggest that nudiflopenes J-M and other four known compounds showed significant inhibitory effects against NO production comparable to the positive control dexamethasone.

3,4-seco-Isopimarane and 3,4-seco-pimarane diterpenoids from Callicarpa nudiflora

A phytochemical investigation was carried out on the extract of a medicinal plant Callicarpa nudiflora,resulting in the characterization of five new 3,4-seco-isopimarane(1-5)and one new 3,4-seco-pimarane diterpenoid(6),together with four known compounds.The structures of the new compounds were fully elucidated by extensive analysis of MS,1 D and 2 D NMR spectroscopic data,and time-dependent density functional theory(TDDFT)calculation of electronic circular dichroism(ECD)spectra,and DFT calculations for NMR chemical shifts and optical rotations.

海南五指山裸花紫珠乙醇提取物通过Wnt/β-连环蛋白信号通路对鼻咽癌细胞株HONE1抗肿瘤机制的研究

目的研究海南五指山裸花紫珠乙醇提取物(CNE)对鼻咽癌细胞株HONE1的抗肿瘤作用及其分子机制。方法培养HONE1细胞,根据CNE处理的浓度分组为溶剂组(0μg/mL)、CNE组A(10μg/mL)、CNE组B(20μg/mL)、CNE组C(40μg/mL)。采用MTT检测细胞增殖抑制率,Transwell实验检测细胞侵袭能力,双染法测定细胞凋亡,蛋白质免疫印迹(Western blot)法检测Wnt3、β-连环蛋白(β-catenin)蛋白相对表达量水平。结果与溶剂组比较,CNE组A、CNE组B、CNE组C的增殖抑制率均大于溶剂组(P<0.05),且CNE各组组间比较,细胞抑制效果与CNE浓度呈正相关(P<0.05)。与溶剂组比较,CNE组A、CNE组B、CNE组C入侵下室的HONE1细胞数均少于溶剂组(P<0.05),且CNE各组组间比较,侵袭细胞数减少情况与CNE浓度呈负相关(P<0.05)。与溶剂组比较,CNE组A、CNE组B、CNE组C的细胞凋亡率均大于溶剂组(P<0.05),且CNE各组组间比较,细胞凋亡率与CNE剂量呈正相关(P<0.05)。与溶剂组比较,CNE组A、CNE组B、CNE组C的Wnt3、β-catenin蛋白相对表达量均低于溶剂组(P<0.05),且CNE各组组间比较,Wnt3、β-catenin蛋白相对表达量与CNE剂量呈负相关(P<0.05)。结论CNE可以抑制HONE1细胞增殖和侵袭,并促进肿瘤细胞凋亡,其机制可能与降低Wnt/β-catenin通路蛋白表达相关。

裸花紫珠重金属污染特征及其指纹图谱的建立

目的:探究裸花紫珠中的重金属的含量,建立无机元素指纹图谱,评价重金属污染风险。方法:采用微波消解处理样品,电感耦合等离子体质谱法测定裸花紫珠中重金属的含量,采用单因子污染指数和内梅罗污染指数法评价其污染风险等级,绘制无机元素指纹图谱。结果:4批样品采用单因子污染指数评价重金属污染时,1批为重污染,3批为中污染。4批样品的综合污染指数评价结果均为轻污染,结论:4批样品存在不同程度的重金属污染,需要引起重视,研究结果可为裸花紫临床药用安全性提供参考依据。

裸花紫珠艾纳香抑菌洗手液的制备和性能检测

制备一种含有裸花紫珠与滇桂艾纳香成分的抑菌洗手液,并通过其起泡性、黏度、抑菌性评价其性能。其处方由18%脂肪醇聚氧乙烯醚硫酸钠(AES)、9%椰油酰胺丙基甜菜碱(CAB)、3%甘油(丙三醇)、7%裸花紫珠艾纳香提取浓缩液、0.26%10%NaOH溶液和62.74%去离子水组成;该洗手液的起泡性能稳定,黏度与市售洗手液相比无显著差异,对金黄色葡萄球菌的抑菌圈直径为22 mm,该洗手液性能良好。

裸花紫珠艾纳香洗衣液的制备和性能检测

提取分离裸花紫珠和滇桂艾纳香的成分,以起泡性、抑菌性、粘度为4个考察因素筛选裸花紫珠艾纳香洗衣液的最优处方,即为11%辛癸基葡糖苷、3.5%椰油酰胺丙基甜菜碱(CAB)、13%月桂酰胺丙基氧化胺、7%十二烷基苯磺酸钠、5%裸花紫珠提取浓缩液、5%艾纳香提取浓缩液、0.7%的柠檬酸和0.3%氯化钠以及54.5%去离子水组成;该洗衣液的起泡性能稳定,对金黄色葡萄球菌的抑菌圈直径为22 mm,去污力能满足人们的需求。

中兽药裸花紫珠颗粒抗菌、抗炎及抗氧化活性研究

为了评估中兽药裸花紫珠颗粒的抗菌、抗炎及抗氧化的效果,试验通过测定裸花紫珠颗粒对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度(MIC)评估其抗菌效果;以脂多糖(LPS)诱导小鼠巨噬细胞(RAW264.7细胞)构建炎症模型,用100μg/mL、50μg/mL、25μg/mL裸花紫珠颗粒药液干预细胞4h,测定药物对一氧化氮(NO)含量和肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)mRNA相对表达量的影响;以H_(2)O_(2)诱导犬脂肪间充质干细胞(cADSC)构建氧化损伤模型,通过试剂盒测定超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,评估药物的抗氧化效果。结果表明:裸花紫珠颗粒对大肠杆菌的MIC为12.5 mg/mL,对金黄色葡萄球菌的MIC为6.25 mg/mL。与模型组相比,各药物组均可显著降低NO含量和TNF-αmRNA的相对表达量(P<0.05);25μg/mL组可显著降低IL-1βmRNA的相对表达量(P<0.05);各药物组的SOD活性显著提高(P<0.05),100μg/mL组的CAT活性显著提高(P<0.05)。说明中兽药裸花紫珠颗粒具有良好的抗菌、抗炎及抗氧化作用。

裸花紫珠醇提物UPLC指纹图谱建立及抗呼吸道合胞病毒谱效关系研究

目的建立裸花紫珠醇提物的指纹图谱,分析其体外抗呼吸道合胞病毒(RSV)活性并进行谱效关系研究。方法以10%、30%、50%、70%、90%乙醇为溶剂,制备4批裸花紫珠药材的20批醇提物。采用超高效液相色谱法,利用《中药色谱指纹图谱相似度评价系统(2012版)》绘制20批裸花紫珠醇提物的指纹图谱并进行相似度评价;采用致细胞病变效应法和MTT法考察裸花紫珠醇提物对RSV的体外抑制活性;运用Pearson相关性分析、灰色关联度分析、正交偏最小二乘(OPLS)分析进行谱效关系研究。结果20批裸花紫珠醇提物有25个共有峰,相似度为0.912~0.998,共有峰峰面积的RSD为33.54%~162.28%。20批裸花紫珠醇提物对RSV的平均半数抑制浓度为9.55~272.23μg/mL;Pearson相关性分析、灰色关联度分析、OPLS分析结果表明,共有峰8、10、12、16、18~19、22~24与药效指标的Pearson相关系数(P<0.05)和回归系数均为负值,关联度均大于0.6,变量重要性投影值均大于1。结论20批裸花紫珠醇提物成分相似但含量差异明显,且具有不同程度的体外抗RSV活性,其中共有峰8、10、12、16、18~19、22~24对应成分可能是裸花紫珠抗RSV的特征成分。

裸花紫珠苯乙醇苷-纳米银复合物制备

目的制备裸花紫珠苯乙醇苷-纳米银复合物。方法以提取液与硝酸银溶液体积比、硝酸银溶液浓度、反应温度、反应时间为影响因素,吸光度为评价指标,单因素试验优化制备工艺。采用紫外-可见分光光度法确定最大吸收波长,在透射电镜下观察外观形态,测定粒径、Zeta电位,红外光谱法分析官能团结构。结果最佳条件为提取液与硝酸银溶液体积比1∶10,硝酸银溶液浓度1 mmol/L,反应温度60℃,反应时间90 min,3批样品吸收曲线稳定。所得纳米银复合物被灰色物质包裹,最大吸收波长为420 nm,平均粒径为26 nm,Zeta电位为^(-1)5.1 mV。在红外光谱图中,1082、535 cm^(-1)处吸收峰减弱,986、861 cm^(-1)处吸收峰消失,1384 cm^(-1)处吸收峰增强。结论该方法稳定可行,可用于制备裸花紫珠苯乙醇苷-纳米银复合物。

裸花紫珠颗粒对罗非鱼源无乳链球菌体外抑菌活性研究

无乳链球菌是导致罗非鱼链球菌病的主要病原菌。为探讨裸花紫珠颗粒对无乳链球菌体外抑菌活性,采用肉汤二倍稀释法测定裸花紫珠颗粒对无乳链球菌的最低抑菌浓度(MIC)和最小杀菌浓度(MBC),同时进行标准无乳链球菌和临床分离无乳链球菌对常用抗菌药的敏感性试验。结果表明,裸花紫珠颗粒对罗非鱼源无乳链球菌的最低抑菌浓度(MIC)为10.000 mg/mL,最小杀菌浓度(MBC)为20.000 mg/mL;药敏试验结果显示临床分离株无乳链球菌对四环素、环丙沙星、氟苯尼考的敏感性降低。该结果为下一步开展裸花紫珠对罗非鱼临床保护试验研究奠定了基础。