Development of a Total Flavonoids Extract of Artemisia Rupestris L.via Nanotechnology and Its Antiviral Effect In Vitro

Objective:We aimed to develop a novel Artemisia rupestris L.flavonoid nano-encapsulation(AFN)preparation and evaulate its anti-hepatitis B virus(HBV)activityin vitro.Methods:First,the AFN was prepared using polylactic-co-glycolic acid(PLGA).Then,after verification of the AFN,in vitro anti-virus assays were conducted by:(1)assessing the inhibitory effect of AFN on the secretion of hepatitis B surface antigens(HBsAg),hepatitis B e-antigens(HBeAg),and the replication of HBV DNAin HepG2.2.15 cells;(2)analyzing the influence of AFN on the activation rate of NF-κB positive cells;and(3)evaluating the effect of AFN on the function of glutathione peroxidase(GSH-PX)enzymes located onthe HepG2.2.15 cell membrane.Results:Compared to the original total flavonoids extract of Artemisia rupestris L.(withoutnano-encapsulation),AFN preparation under the maximum nontoxic concentration effectively inhibited the secretion of HBsAg,HBeAg,and HBV DNA from HepG2.2.15 cells.At the same time,AFN preparation promoted not only the activation rate of NF-κB positive cells,but also antiviral GSH-PX enzyme function.In conclusion,nano-encapsulation of the flavonoids extract of Artemisia rupestris L.showed an enhanced anti-HBV effect in vitro compared to the original total flavonoids extract(without nanoencapsulation);therefore,nano-encapsulation has great potential for the development of a novel antiviral herbal medicine preparation with improved efficacy.

HPLC法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D的含量

目的建立高效液相色谱(HPLC)法同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草黄素、猫眼草酚D的含量。方法采用Agilent ZORBAX SB-C18(250 mm×4.6 mm,5μm)色谱柱;流动相:乙腈(A)-纯水(B),梯度洗脱;体积流量:0.8 mL·min^(-1);柱温:30℃;进样量:10μL;检测波长:210 nm。结果青蒿素、青蒿乙素、猫眼草黄素和猫眼草酚D进样量分别在1.6088~16.088μg(r=0.9999)、0.0141~0.1414μg(r=1)、0.1851~1.8509μg(r=0.9999)、0.1441~1.4414μg(r=0.9999)范围内与峰面积呈良好的线性关系,平均加样回收率(n=6)分别为102.44%、97.82%、95.07%、95.55%,RSD分别为1.12%、1.44%、1.29%、1.53%。结论该方法简便准确、可靠稳定、重复性好,可用于同时测定黄花蒿水提浸膏中青蒿素、青蒿乙素、猫眼草酚D、猫眼草黄素的含量。

椒蒿精油成分随生长时间的OPLS-DA分析

为探究不同生长时间椒蒿精油品质的差异性,采用气相色谱-质谱(GC-MS)联用技术对椒蒿精油的化学成分进行分析,通过正交偏最小二乘法判别分析(OPLS-DA)和聚类热图分析验证生长时间对其化学成分及相对含量的影响。GC-MS分析结果表明,各生长时间共有物质25种,其中13种萜烯类、9种醇类、3种其他物质,除此之外在个别生长时间还检测出酯类、脂肪酸及酮类等物质。聚类热图分析表明,大多数物质含量在采摘时间S1和S2较多,S3和S4次之,S5和S6最低。萜烯类物质整体呈现先下降后上升再下降的趋势,萜烯类物质总的相对含量在4月份相对偏高,5月份次之,6月份最低。醇类物质在S4之前各生长时间含量相对较多,S4之后相对含量变少,同时α-生育酚在S4之前含量也较多。变量投影重要性(VIP)分析表明,共有5种物质的VIP大于1,分别为榄香素、α-蒎烯、3-蒈烯、γ-谷甾醇、β-谷甾醇。因此,5月下旬之前采摘椒蒿并提取精油,得到的椒蒿精油品质相对较好。

黄花蒿(Artemisia annua L.)提取物对两种病原真菌的生物活性

以石油醚Ⅰ(30℃～60℃)、石油醚Ⅱ(60℃～90℃)、乙醇和丙酮等4种溶剂对黄花蒿的根、茎、叶进行初步提取,采用生长速率法测定不同提取物对玉米小斑病菌、棉花枯萎病菌的抑菌活性。结果表明,黄花蒿提取物对玉米小斑病菌的生物活性好于棉花枯萎病菌;黄花蒿叶的提取物抑菌效果最好,根的抑菌效果最差;叶的石油醚(60℃～90℃)提取物对玉米小斑病菌EC50为156.32mg/L;而叶的丙酮提取物对玉米小斑病菌的EC50为82.37mg/L。

<i>Artemisia vulgaris</i>L. Chemotypes

Artemisia vulgaris L. was collected from various regions in the USA and Montreal Canada. Gas Chromatography-Mass Spectrometry was used to identify the analytes present in the volatiles extracted by headspace solid-phase microextraction of the crushed leaves and flowers. Four distinct chemotypes are were found: One featuring the coexistence of ar-curcumene and α-zingiberene;two marked by the presence or absence of thujone and santolinatriene;and a fourth characterized by the presence of crysanthenyl acetate (40%). DNA was used to confirm the identity of Artemisia vulgaris L.

Effect of Artemisia annua L. as Substitute to Sulfonamides (Sodium Sulfadimerzine) on Coccidiosis and Growth Performance in Rabbits

Coccidiosis is a disease caused by protozoa of the genus Eimeria which seriously affects young rabbits. Treatment based on the use of anticoccidial drugs is increasingly ineffective due to the rapid emergence of resistant strains of coccidia and the high cost of drugs. Consumer demand for rabbit products without chemical residues led to a growing interest in the use of medicinal plants as an alternative treatment for coccidiosis. The present study was carried out during the period of August to December 2020 to assess the anticoccidial effect of hydro-ethanolic extract of leaves of Artemisia annua L., in young rabbits. The antiparasitic efficacy of Artemisia extract was tested on 15 young rabbits (whose age varied between 7 and 9 weeks) divided into 5 lots of 3 animals. The average weight of these animals was 790 g. The results of this study show that the feces samples and the weight of young rabbits before administration of the treatment and the coprological examination (every 7 days for 4 weeks) show a fecal excretion reduction rate (FECRT) of 55.13% in the lot treated by sulfonamide. On the other hand, in animals received treatments extract of the leaves of Artemisia annua L., the average FECRT is evaluated at 69.64%, 79.22%, and 96.36% for respective doses of 400, 800 and 1200 mg/kg bodyweight and proves their anticoccidial effect. Furthermore, the variation in mean Eggs Per Gram (EPG) of coccidia and the average weekly weight gain (AWWG) of each lot were significant in the lots treated with hydro-ethanolic extract (P 0.05). The greatest reductions in oocystal excretion and weight gain obtained were those of lot 5, treated at 1200 mg/kg of hydro-ethanolic leaves extract of Artemisia annua L.

Relationship between the haplotype distribution of Artemisia halodendron(Asteraceae) and hydrothermal regions in Horqin Sandy Land, northern China

The genetic diversity of Artemisia halodendron(Asteraceae), a constructive and dominant species in Horqin Sandy Land,was investigated to examine the genetic relationships with different hydrothermal regions in Horqin Sandy Land. We sequenced chloroplast DNA(cp DNA) fragments(trn L–F) of 243 plants from 10 populations across the Horqin Sandy Land.The analyses of cp DNA variation identified seven haplotypes. A low level of haplotype diversity(H_d=0.706) and nucleotide diversity(π=0.0013) was detected. Haplotypes clustered into two tentative clades. Low genetic differentiation among regions was consistently indicated by hierarchical analyses of molecular variance(AMOVA). Across the sampled populations, the haplotype distributions were differentiated with hydrothermal gradients.

Study on Allergic Rhinitis of Artemisia Sphaerocephala

Objective:To explore the cause and feasibility of Artemisia ordosica allergic rhinitis.Methods:The main allergic causes of allergic rhinitis patients in Chifeng City of inner Mongolia were analyzed by a control method.Results:According to the results,Artemisia ordosica is the most important cause of allergic rhinitis in inner Mongolia.In addition,allergic history,asthma and other diseases,or animal hair and other factors can also increase the risk of patients with allergic rhinitis.Conclusions:Artemisia sphaerocephala is the main cause of allergic rhinitis.In recent years,allergic rhinitis has attracted the attention of international organizations,and the medical community has also studied more methods for the treatment of rhinitis,such as putting forward relevant traditional Chinese medicine(TCM)treatment methods.

黄花蒿对大蒜化感作用研究

为探究黄花蒿对大蒜鳞茎萌发及其幼苗生长的化感作用,用水提取黄花蒿叶、根、根际土壤,获取不同质量浓度的浸提液,用于处理大蒜鳞茎,测定大蒜鳞茎萌发率及其幼苗生长状况和叶绿素含量等指标。结果表明:随着黄花蒿水浸液浓度的升高,黄花蒿水浸液对大蒜鳞茎萌发率及其幼苗苗高、根长、叶绿素a和b含量、叶绿素含量、过氧化物酶活性、植株鲜质量、植株高的抑制作用不断增大,对大蒜幼苗丙二醛含量的促进作用不断增强;黄花蒿水浸液通过抑制大蒜鳞茎萌发率及其幼苗过氧化物酶活性、增加膜脂过氧化和逆境伤害、抑制叶绿素a、b和叶绿素的生成,从而抑制植株生长,表现出化感作用,且化感效应随着黄花蒿水浸液浓度的升高而增大。本研究为农作物和药用植物栽培及植物间化感作用研究提供参考。

新疆一枝蒿不同部位水提物的体外抗炎效果

为了探究新疆一枝蒿不同部位水提物的体外抗炎效果,本试验在体外用脂多糖(LPS)诱导小鼠单核巨噬白血病细胞(RAW264.7)构建炎症模型,采用水提法对新疆一枝蒿不同部位(叶、茎、花)进行提取,运用噻唑蓝(MTT)法检测新疆一枝蒿不同部位水提物对细胞活力度的影响,实时荧光定量PCR(RT-qPCR)检测其对炎症相关基因TNF-α、IL-1β和IL-6 mRNA表达的影响,Western blot检测其对TNF-α、IL-1β和IL-6蛋白表达的影响。结果显示,新疆一枝蒿叶、茎、花水提物分别在0.078~1.250、0.078~5.000和0.078~1.250 mg/mL的浓度范围内对RAW264.7细胞活力度无显著抑制作用。与LPS模型组相比,新疆一枝蒿叶、茎、花水提物浓度范围在0.313~1.250、0.625~2.500和0.625~1.250 mg/mL时,细胞活力度分别提高了23.12%~28.14%(P<0.01)、10.88%~18.62%(P<0.01)和16.13%~16.76%(P<0.05)。浓度为1.250 mg/mL的叶水提物可以极显著降低TNF-α、IL-1β和IL-6的mRNA和蛋白表达(P<0.01);浓度为0.625和1.250 mg/mL的茎水提物可以极显著降低IL-1β的mRNA和蛋白表达(P<0.01),以及IL-6和TNF-α的mRNA表达(P<0.01);浓度为0.625 mg/mL的花水提物可以极显著降低IL-1β和IL-6的mRNA和蛋白表达(P<0.01)。结果表明,新疆一枝蒿不同部位(叶、茎、花)水提物均有抗炎效果,本试验结果可为新疆一枝蒿的兽医临床应用提供参考依据。

黄花蒿中青蒿素含量的RP-HPLC法测定

建立黄花蒿中青蒿素的高效液相色谱测定方法,采用柱前衍生RP-HPLC法测定黄花蒿中青蒿素的含量,采用ZORBAXXDB-C18(4.6mm×150mm,5μm)色谱柱,甲醇-0.01mol/L醋酸钠-醋酸缓冲液(pH5.8,体积比62∶38)为流动相;检测波长:260nm;流速:0.8mL/min;柱温:30℃。对广西、沈阳、北京、郑州、苏州和杭州等不同产地的野生黄花蒿样品、以及同一产地不同采集时间黄花蒿样品进行检测,结果表明不同地区青蒿素含量差异很大,同一地区不同采集时间黄花蒿样品的青蒿素含量也有差异。该法准确可靠、重现性好,能准确地反映青蒿素含量的检测。

不同采收期和种植地貌青蒿的品质分析

目的:比较不同采收期喀斯特山地和传统平地上种植青蒿的表型特征和有效成分含量变化,确定青蒿的最适种植环境及最佳采收期。方法:采用直接测量法、高效液相色谱法和比色法,分别测定2种地貌上种植的5个采收日期青蒿的表型特征,以及青蒿素、青蒿酸、总黄酮和总多酚含量。结果:同一采收日期,在喀斯特山地上种植的青蒿呈全株矮且宽、根系稀疏且为浅根系的生长状态,而在传统平地种植的青蒿呈枝繁叶茂、株高茎壮、根系茂密且为深根系的生长状态;传统平地种植的青蒿总黄酮、总多酚、青蒿素、青蒿酸含量高于喀斯特山地种植的青蒿。喀斯特山地和传统平地上种植青蒿的总黄酮、青蒿素、青蒿酸含量均在7月末最高,总多酚含量的变化趋势与青蒿表型特征的变化趋势相似。结论:传统平地上种植的青蒿表型特征及青蒿素等成分含量的最高值均优于喀斯特山地上种植的青蒿,7月下旬为青蒿的最佳采收期。

RP-HPLC法测定青蒿中青蒿素的含量

目的建立青蒿(黄花蒿)干叶中青蒿素的含量测定方法,为甲醇提取青蒿素的工业化生产工艺提供依据。方法采用正交试验设计,优选测定条件,采用Shimadzu Shim-pack VP-ODS C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(55∶45),流速为1.0 mL.min-1,检测波长为210 nm。结果最佳的含量测定条件为加40倍量甲醇,超声提取3次,每次50 min。青蒿素在1～40μg范围内与峰面积呈良好的线性关系(r=0.99998),平均加样回收率为99.8%,RSD为1.0%(n=6)。结论该方法简便、准确,适用于黄花蒿中青蒿素的含量测定。

pH-时滞型新疆一枝蒿结肠定位片的制备及体外释放评价

目的:制备pH-时滞型新疆一枝蒿结肠定位片并对其体外释放性能进行评价。方法:采用压制包衣、喷雾包衣的方法制备结肠定位片,以总黄酮累积释放度为指标,分别考察压制包衣时滞材料处方比例、包衣用量、上下层填料比例,pH控释层处方比例与包衣增重。结果:经过考察,最优工艺处方为EC-PEG 6 000(4∶1),压制包衣0.1 g,压制包衣上下层填料(1.5∶1),Eudragit FS 30 D-Eudragit L30 D-55(2∶1),p H控释层包衣增重为7%时,制得结肠定位制剂在0.1 mol/L HCl溶液中2 h内不释药,在pH 6.8磷酸盐缓冲液中4 h累积释放度<10%,在p H 7.5磷酸盐缓冲液中6 h累积释放度>80%。结论:经过筛选后的结肠定位片能够实现结肠定位释放的目的且体外释放稳定。

黄花蒿中青蒿素的柱前衍生HPLC-UV分析

研究衍生化条件对HPLC-UV法测定黄花蒿青蒿素含量的影响。建立了黄花蒿青蒿素的柱前衍生HPLC-UV测定方法:即青蒿素标准品溶液(或黄花蒿提取浸膏)以体积分数为95%乙醇补充(溶解)至15.0mL,再加入质量分数为0.2%的NaOH8.0mL,45℃水浴30min后冷却至室温,加入0.08mol/L的HAc10mL摇匀,以95%乙醇定容至50mL容量瓶中;HPLC分离,260nm紫外检测。所得方法检测线性范围为0～200mg/L,相对标准偏差(RSD)值为1.40%,平均回收率(n=5)为100.1%。用该方法测定了黑龙江引种的黄花蒿提取物样品,稳定性RSD值为4.35%,重现性RSD值为4.96%。证明该方法简单、稳定、重现性好。

青蒿挥发油的化学成分、生物活性及应用的研究进展

目的对近20年有关青蒿挥发油的化学成分、生物活性以及应用领域的研究进行了全面、系统的综述,为充分利用青蒿挥发油提供参考。方法在Web of Science、Scifinder、PubMed、中国知网和万方等数据库,对2001—2023年的相关文献进行检索,以青蒿挥发油、提取方法、活性、应用等为关键词进行检索,对化合物的种类、活性等进行归纳总结。结果青蒿挥发油的化学成分主要包括倍半萜类、醇类、醛类、酸类、酚类、酯类、酮类,其具有抗炎、抑菌、抗寄生虫、抗病毒、抗肿瘤等生物活性。结论青蒿挥发油呈多样性的生物活性,其中抗菌作用较为显著,可以作为化妆品的祛屑成分,其作用机制还需进一步深入研究。

青蒿查尔酮合成酶(AaCHS)基因家族鉴定及光调控分析

[目的]为了解青蒿(Artemisia annua L.)查尔酮合成酶(chalcone synthase,CHS)基因家族的分子进化特性及其在不同组织部位的表达情况。[方法]从Pfam数据库下载CHS蛋白HMM模型,并使用HUMMER 3.0、Blastp和CD-search鉴定出青蒿基因组中的CHS基因家族成员。采用TBtools、EXPASy、CELLO v.2.5、MEGA 7.0、MEME、SOPMA、SWISSMODEL和PlantCARE等软件对其氨基酸与碱基序列进行生物信息学分析,通过8个不同组织部位的转录组数据对AaCHS基因家族成员进行表达分析。[结果]青蒿基因组中共有16个AaCHS基因家族成员,蛋白结构分析显示AaCHS蛋白质结构并不稳定,将其分为4个组,基因结构分析显示所有AaCHS基因蛋白均具有外显子,数量为2~4,内含子数量为0~2,其中8个AaCHS基因不具有内含子。启动子顺式作用元件分析表明该基因家族成员启动子上含有大量光响应、植物激素响应元件;CD-search验证结果发现每个AaCHS蛋白均有Chal-sti-synt_N结构域。[结论]AaCHS家族成员表达分析结果表明,16个AaCHS基因在不同组织和光处理下表达不同。GO富集结果得到GO二级分类的生物过程和分子功能,生物过程共富集了80条序列;细胞组分共富集了19条序列,其中二级分类过程细胞质中富集了11条序列,数量最多,与亚细胞定位预测结果一致;推测AaCHS基因在细胞质中调控黄酮类化合物的累积。通过青蒿光调控差异表达分析,推测AaCHS7、AaCHS10在青蒿受到红光光处理中起正向调控作用。

衍生化条件的优化及不同产地青蒿中青蒿素含量的柱前衍生化-HPLC法测定

Derivation conditions of pre-column derivation-HPLC method used for determinating artemisinin content were compared and selected,and artemisinin content in above-ground part of Artemisia annua L.from seventeen locations was compared by optimal pre-column derivation-HPLC method.The optimal derivation condition is selected via comparing of 0.2% NaOH solution addition(3,4,5,6 and 7 mL),derivation temperature(30 ℃,35 ℃,40 ℃,45 ℃ and 50 ℃) and derivation time(0,2,5,10,20,40 and 60 min) during derivation process.The optimal derivation condition is adding 5 mL 0.2% NaOH solution,then insulating in 40 ℃ water bath for 10 min.Differences of artemisinin content in A.annua from seventeen locations are obvious,the general trend is that artemisinin content decreases gradually with location from south to north,in which artemisinin content in A.annua from Youyang of Chongqing is the highest with a value of 7.08 mg·g-1.The method is simple and accurate with good reproducibility,and can be used to determinate artemisinin content in medicinal material of A.annua.

中草药青蒿(Artem isia annua L.)的实时荧光PCR检测

中草药青蒿,学名黄花蒿(Artem isia annuaL.),属于中国独有的抗疟药用植物资源之一。根据蒿属植物rDNA ITS序列多态性设计TaqM an探针及引物,通过实时荧光PCR方法对黄花蒿进行检测。结果表明:该组探针-引物对黄花蒿有很强的特异性,除黄花蒿外,其余8种对照蒿属材料均未检测到荧光信号。该方法快速、简便、安全、准确,适用于黄花蒿的出入境检验。

新疆一枝蒿口服液的制备工艺优化研究

为了探究新疆一枝蒿口服液的最佳制备工艺,采用水提醇沉法提取新疆一枝蒿中的总黄酮,经单因素、响应面试验获取最佳的制备工艺参数,并通过薄层层析与含量限度测定对新疆一枝蒿口服液建立初步的质量控制。结果表明:新疆一枝蒿最佳制备工艺为料液比1∶14,煎煮时间为1.5 h,醇沉浓度为50%。供试品色谱中,在与对照品色谱相应的位置上,显相同颜色的斑点,含量限度测定黄酮含量范围在28.67~29.59 mg/mL,平均值为29.10 mg/mL,最低限度为23.28 mg/mL。