Studies on Acaricidal Bioactivities of Artemisia annua L.Extracts Against Tetranychus cinnabarinus Bois.(Acari:Tetranychidae)

The aim of this study was to determine the best extraction technique, the most suitable solvent, the optimal plant parts, and the acaricidal activities of Artemisia annua L. The petroleum ether （30-60℃）, petroleum ether （60-90℃）, ethanol, acetone, and water parallel and sequenced extracts were obtained from the leaves, stems and roots of different period of A. annua L. in April, May, June, July and September respectively. And then the acaricidal bioactivities against Tetranychus cinnabarinus of all extracts were determined by the slide-capillary method in the laboratory. The results indicated that the acaricidal bioactivities elevated as the development of A. annua plant at the concentration of 5 mg mL-L The general tendency exhibited the sequence of July 〉 June 〉 May 〉 April, but September decreased comparing to July. However, the most effective extracts in five months were all acetone parallel extract of A. annua leaf, and the corrected mortalities treated after 48 h ranged from 74 to 100%. The median lethal concentrations （LC50） against T. cinnabarinus of acetone parallel extracts ofA. annua leaves in September, July, June, May and April were 0.5986, 0.4341, 0.8376, 0.9443 and 1.3817 mg mL^-1, respectively, treated after 48 h. The 13 groups were isolated from acetone extracts ofA. annua leaves in July by column chromatography, both the 1 lth and 12th groups exhibited strong bioactivities. The median lethal concentrations of the 1 lth and 12th groups against T. cinnabarinus were 0.3683 and 0.1586 mg mL^-1, respectively. The acetone parallel extract ofA. annua leaf in July was the most toxic to T. cinnabarinus and the corrected mortality was 100% after 48 h. The acetone parallel extract of the 1 lth and 12th groupswere the most active components, acted as the emphases in further study.

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.

The AaBBX21–AaHY5 module mediates light-regulated artemisinin biosynthesis in Artemisia annua L.

The sesquiterpene lactone artemisinin is an important anti-malarial component produced by the glandular secretory trichomes of sweet wormwood(Artemisia annua L.).Light was previously shown to promote artemisinin production,but the underlying regulatory mechanism remains elusive.In this study,we demonstrate that ELONGATED HYPOCOTYL5(HY5),a central transcription factor in the light signaling pathway,cannot promote artemisinin biosynthesis on its own,as the binding of AaHY5 to the promoters of artemisinin biosynthetic genes failed to activate their transcription.Transcriptome analysis and yeast two-hybrid screening revealed the B-box transcription factor AaBBX21 as a potential interactor with AaHY5.AaBBX21 showed a trichome-specific expression pattern.Additionally,the AaBBX21–AaHY5 complex cooperatively activated transcription from the promoters of the downstream genes AaGSW1,AaMYB108,and AaORA,encoding positive regulators of artemisinin biosynthesis.Moreover,AaHY5 and AaBBX21 physically interacted with the A.annua E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1(COP1).In the dark,AaCOP1 decreased the accumulation of AaHY5 and AaBBX21 and repressed the activation of genes downstream of the AaHY5–AaBBX21 complex,explaining the enhanced production of artemisinin upon light exposure.Our study provides insights into the central regulatory mechanism by which light governs terpenoid biosynthesis in the plant kingdom.

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。

Studies on the biosynthesis of arteannuin——Ⅳ.The biosynthesis of arteanniuin and arteannuin B by the leaf homogenate of Artemisia annua L.

Arteannuic acid and(-)11R,13-d(?)ydroarteannuic acid are the key intermediates in the biosynthesis of arteannuin by the leaf homogenate of Artemisia annua L.,and arteannuic acid and epoxyarteannuic acid are the intermediates of arteannuin B.and epoxyarteannuic acid can not be transformed into arteannuin by the homogenate.

Transcriptional regulation of artemisinin biosynthesis in Artemisia annua L.

Artemisinin, also known as qinghaosu, a sesquiterpene endoperoxide lactone isolated from the Chinese medicinal plant Artemisia annua L., is the most effective antimalarial drug which has saved millions of lives.Due to its great antimalarial activity and low content in wild A. annua plants, researches focused on enhancing the artemisin yield in plants became a hotspot. Several families of transcription factors have been reported to participate in regulating the biosynthesis and accumulation of artemisinin.In this review, we summarize recent investigations in these fields, with emphasis on newly identified transcription factors and their functions in artemisinin biosynthesis regulation, and provide new insight for further research.

Study on the Repairing Effect of Cosmetics Containing <i>Artemisia annua</i>on Sensitive Skin

In view of the increasing sensitivity of consumer skin in recent years, cosmetics containing Artemisia annua extract was tested to evaluate its effectiveness in repairing sensitive skin. Through the experiment of xylene-induced ear swelling in mice, it was found that the inhibition rates of ear swelling in mice induced by xylene in three groups of cosmetics containing Artemisia annua extract reached 60.40%, 73.36% and 74.01%, respectively, close to the positive drug group. Twenty-five sensitive skin volunteers were selected for human clinical trial, and the skin TEWL value, cuticle hydration degree and skin heme (ultra-high concentration) were tested. The results showed that using cosmetics containing Artemisia annua extract for four weeks could effectively increase the hydration degree of cheek cuticle by 63.90% and reduce transepidermal waterloss (TEWL) by 21.51%. The skin heme (ultra-high concentration) decreased by 69.14% and the affected area decreased by 77.47%. The results show that the cosmetics containing Artemisia annua extract can inhibit inflammation, repair skin barrier, improve damaged skin, and reduce redness and other sensitive skin symptoms.

Effects of different doses of cadmium on secondary metabolites and gene expression in Artemisia annua L.

This study aims to elucidate the underlying molecular mechanisms of artemisinin accumulation induced by cadmium （Cd）. The effects of different Cd concentrations （0, 20, 60, and 120 μmol/L） on the biosynthesis ofArtemisia annua L. were examined. Intermediate and end products were quantified by HPLC-ESI- MS/MS analysis. The expression of key biosynthesis enzymes was also determined by qRT-PCR. The results showed that the application of treatment with 60 and 120 μmol/L Cd for 3 days significantly improved the biosynthesis of artemisinic acid, arteannuin B, and artemisinin. The concentrations of artemisinic acid, arteannuin B, and artemisinin in the 120 llmol/L Cd-treated group were 2.26, 102.08, and 33.63 times higher than those in the control group, respectively. The concentrations of arteannuin B and artemisinin in 60 μmol/L Cd-treated leaves were 61.10 and 26.40 times higher than those in the control group, respectively. The relative expression levels of HMGR, FPS, ADS, CYP71AV1, DBR2, ALDH1, and DXR were up-regulated in the 120 μmol/L Cd-treated group because of increased contents of artemisinic metabolites after 3 days of treatment. Hence, appropriate doses of Cd can increase the concentrations of artemisinic metabolites at a certain time point by up-regulating the relative expression levels of key enzyme genes involved in artemisinin biosynthesis.

The Effect of Different Doses of Composite Artemisia annua Extract on Ruminal Fermentation of Dairy Goats in vitro

An in vitro study was conducted to determine the effect of different doses of Composite Artemisia annua extract(CAE) on rumen microbial fermentation.CAE was a crude extract from the stem and leaves of Composite Artemisia annua extracted with ethanol.The doses of CAE supplementation with frozen-dry form were:control(no addition)

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。

Study on Soothing and Oil Control Anti-acne Effects of Artemisia Annua Extract

Preliminary exploration of the soothing,oil control,acne removing effects and its mechanisms of Artemisia annua extract.The soothing effect of Artemisia annua extract was tested by hyaluronidase biochemical reaction.The soothing and oil-controlling effects were investigated by cell model.The inhibitory effect on propionibacterium acnes was studied by suspension quantification.The results showed that Artemisia annua extract could effectively inhibit the degradation of hyaluronic acid(P<0.01).Artemisia annua extract significantly inhibited the secretion of inflammatory cytokines TNF-αand IL-6 in RAW264.7 cells(P<0.05).Artemisia annua extract at 0.5%,0.25%,0.125%could significantly inhibit the secretion of oil by SZ95 cells(P<0.05).The minimum inhibitory concentration of Artemisia annua extract against propionibacterium acnes was 0.625%,and the inhibitory rate against propionibacterium acnes increased with the increase of the concentration of Artemisia annua extract.In summary,Artemisia annua extract can achieve acne efficacy through soothing and oil control,and this function may be achieved by reducing hyaluronic acid degradation,inhibiting inflammatory pathways produced by inflammatory factors TNF-αand IL-6,inhibiting oil secretion,and inhibiting the growth of propionibacterium acnes.

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。

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。

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。

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.

黄花蒿提取物生产工艺条件及其对乳中共轭亚油酸(CLA)生成的调控作用

黄花蒿乙醇提取物含有多种营养活性物质,主要包括萜类、倍半萜类、芳香族类、脂肪酸类、甾体类、黄酮类、生物碱类等化合物。反刍动物日粮中添加黄花蒿乙醇提取物能够增加其乳中共轭亚油酸(CLA)含量。综述了黄花蒿乙醇提取物的主要营养活性物质、提取工艺条件、营养活性物质的检测方法及黄花蒿提取物在动物日粮中的适宜添加量,旨在为生产CLA乳提供参考。

UPLC-MS/MS法检测青蒿中青蒿素的含量

采用UPLC-MS/MS法检测青蒿中青蒿素的含量,色谱柱Waters ACQUITY BEH C18(50 mm×2.5mm,1.9μm),以甲醇和水梯度洗脱,甲醇-水体积比为40%(0 min)-70%(4.5 min)-40%(5.0 min)-40%(5.5 min),流速0.30 mL/min,柱温30℃,进样量3.0μL,离子模式(ESI+)分析,青蒿素母离子为m/z=283.3。子离子m/z=151.2,m/z=209.4。结果表明,在上述条件下,青蒿素与杂质分离完全,无明显干扰,检出限达到0.174 ng,平均回收率为99.38%,精密度良好,青蒿中青蒿素含量为(1 045.19±27.55)mg/kg,该法适用于青蒿素分析。

黄花蒿bHLH转录因子基因家族鉴定及光调控分析

目的:基于黄花蒿全基因组及转录组数据,进行AabHLH基因家族生物信息学分析及在不同光处理下的基因表达分析。方法:基于基因隐马科夫模型(HMM)对AabHLH基因进行鉴定,利用Pfam、GSDS、MEME等在线分析软件对基因结构、分子进化及保守结构域等进行分析。结果:全基因组水平共鉴定出99条黄花蒿AabHLH基因,包含49对复制基因对;基于系统发育树将其分为13个亚族,所有亚族AabHLH蛋白亚细胞定位于细胞核且均为亲水性蛋白,同一亚族基因具有相似的外显子-内含子结构及保守基序。基因表达模式分析发现,AabHLH基因在不同水平上响应蓝光、红光、远红光、白光调控,推测其中6个亚族基因为光照条件下,7个亚族基因为非光照条件下青蒿素合成途径调控基因。结论:在全基因组水平鉴定了AabHLH基因,为深度解析AabHLH基因功能及其对光调控下青蒿素生物合成调控机制提供参考。

青蒿UPLC指纹图谱的建立与多种成分含量测定

目的:建立30批青蒿样品的UPLC指纹图谱及有机酸类、黄酮类、香豆素类等成分的含量测定方法。方法:采用Agilent Eclipise Plus C18(50 mm×2.1 mm,1.8μm)色谱柱;以0.1%磷酸溶液-甲醇为流动相,梯度洗脱;建立指纹图谱;并对青蒿中有机酸类、黄酮类、香豆素类等9个成分进行含量测定。结果:建立了青蒿UPLC指纹图谱及同时测定多成分含量的方法。9种成分在测定范围内线性关系良好,不同产地的青蒿药材质量存在一定差异。江苏盱眙的青蒿综合评价质量最优。结论:本实验建立的方法准确可靠,可同时测定青蒿中东莨菪内酯、滨蒿内酯、异绿原酸A、异绿原酸B、异绿原酸C、木犀草素、芹菜素、猫眼草黄素、艾黄素的含量。

HPLC-ELSD法测定黄花蒿提取物中青蒿素

目的 建立HPLC-ELSD法对黄花蒿提取物中青蒿素的测定方法。方法 采用HPLC-ELSD法。色谱柱：Inertsil ODS-3（150mm×4．6mm，5μm）；流动相为乙腈-水（69：31，用三氟醋酸调pH至3）；体积流量：1．0mL／min；检测器漂移管温度：45C；载气体积流量：1．0L／min；分流：开；柱温：25℃。结果 青蒿素在0．5893～5．8935mg／mL时面积的常用对数值与其质量浓度的常用对数值线性关系良好（r=0．989）；样品平均回收率为98．71％（RSD为0．42％，n=6）。结论 该方法灵敏、准确可靠，可作为黄花蒿提取物中青蒿素的定量方法。