Rehmannia glutinosa

地黄
  • 文章类型: English Abstract
    地黄是一种常用的中药材,具有清热活动,血液冷却,滋阴,和促进体液。环烯醚萜苷是主要的生物活性。环烯醚萜氧化酶是环烯醚萜苷生物合成途径中的关键限速酶。在这项研究中,基于转录组数据筛选了环烯醚萜类氧化酶基因RgIO,其次是生物信息学分析,表达特征检测,和亚细胞定位分析。结果表明,RgIO的编码区为1536bp,编码511个氨基酸,分子量约为58258。01.RgIO的蛋白质序列含有细胞色素P450氧化酶的保守结构域和基序。RgIO在Strigaasiatica中与其直系同源蛋白具有最高的序列同一性,Strigahermonthica,和Centrantheragrandiflora,并具有良好的序列同一性(77。28%)与长春花CrIO。RgIO在凝集素的叶中显示出特异性表达。为了回应MeJA归纳法,MeJA在处理后叶和根中的表达量增加了3。15和1。3次在3小时和6小时,分别。亚细胞定位结果表明,RgIO分布在内质网中。农杆菌介导的RgIO基因在谷草叶片中的瞬时表达使梓醇含量增加0。与空载体的瞬时表达比拟82倍。本研究为凝析油的分子调控和生物合成提供了关键的靶基因,为揭示凝析油完整的生物合成途径奠定了基础。
    Rehmannia glutinosa is one of the commonly used Chinese herbal medicines, which has activities of heat-clearing,blood-cooling, Yin-nourishing, and body fluid-promoting. Iridoid glycosides are the main bioactive in R. glutinosa. Iridoid oxidase is a key rate-limiting enzyme in the biosynthetic pathway of iridoid glycosides. In this study, an iridoid oxidase gene Rg IO was screened based on the transcriptome data, followed by bioinformatics analysis, expression characteristic detection, and subcellular localization analysis. The results show that the coding region of Rg IO is 1 536 bp, with 511 amino acids encoded, and the molecular weight is about 58 258. 01. The protein sequence of Rg IO contains the conserved domains and motifs of cytochrome P450 oxidases. Rg IO has the highest sequence identities with its ortholog proteins in Striga asiatica, Striga hermonthica, and Centranthera grandiflora and has good sequence identities(77. 28%) with Catharanthus roseus Cr IO. Rg IO shows specific expression in the leaf of R. glutinosa. In response to MeJA induction, the expression of MeJA in leaves and roots after treatment increases by 3. 15 and 1. 3 times at 3 h and 6 h,respectively. The result of subcellular localization shows that Rg IO is distributed in the endoplasmic reticulum. Agrobacterium-mediated transient expression of Rg IO gene in leaves of R. glutinosa makes the content of catalpol increase by 0. 82 times compared with the transient expression of the empty vector. This study provides a key target gene for the molecular regulation and biosynthesis of catalpol in R. glutinosa and lays a foundation for revealing the complete biosynthetic pathway of catalpol.
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  • 文章类型: Journal Article
    地黄(又称毛地黄)是一种多年生双子叶草本植物,在中医药中发挥着重要作用。其活性成分对血液系统具有广泛的药理作用,内分泌系统,免疫系统,心血管系统,和神经系统(Zhang等人。2008).2022年5月,在焦作市(35°01\'44.20″N,113°05\'30.63英寸),河南省,中国的发病率约为54%(约1,300种植物)。不规则的棕色病变最初出现在基叶的尖端,然后进展到整个叶子,导致叶子干燥(柔软。Fig.1-A,B,C).从植物的基部到顶部的叶子中相继出现了相同的症状,最终导致整株植物死亡。为了识别病原体,从八株植物中随机收集了八株有症状的叶子,在病灶边界处切成小块(5×5mm)。这些碎片在75%的乙醇中表面消毒15秒,然后用1%NaClO持续1分钟,在无菌水中冲洗三次,置于25℃黑暗条件下的马铃薯葡萄糖琼脂(PDA)培养基上3天。最后,采用单孢子法获得12株纯化分离物(DHY1-DHY12)。用分生孢子悬浮液(106分生孢子/ml)接种R.glinosa幼苗的叶子,每个分离株接种三株植物。用无菌水处理对照。将所有接种和对照植物在25℃、80±10%湿度和8-h/16-h黑暗/光照循环下孵育。该实验重复三次。5天后,接种DHY4-DHY10的叶片出现与幼苗苗圃中病叶相似的症状,而接种DHY1-DHY3,DHY11-DHY12的植物则无症状(柔软。图1-D,E).从病叶中重新分离出相同的真菌,履行科赫的假设。病原体DHY4到DHY10,显示相似的形态,在形态上被鉴定为曲霉属。(Visagie等人。2014).选择分离物DHY5用于进一步研究。在PDA车牌上,殖民地被白色的绒毛菌丝体覆盖(柔软。图1-F)。分生孢子为o石黄色,向外的同心圆。囊泡呈球形,直径约为20.1-26.6μm(柔软。图1-G)。分生孢子柱壁光滑透明,分生孢子头辐射。分生孢子为浅黄色至橙色,渗出液清澈到橙色的小滴。分生孢子直径为(2.53-3.25)μm×(2.58-3.47)μm(n=50)(柔软。图1-H)。为了进一步的分子鉴定,用引物对ITS1/ITS4和BT2a/BT2b(Glass和Donaldson。1995),分别。ITS(PP355445)和TUB(PP382788)序列的BLASTn搜索显示与A.westerdijkiae(OP237108和OP700424)的相似性为100%和98.42%,分别。基于ITS和TUB的串联序列的系统发育分析证实该真菌是A.westerdijkiae,(柔软。图2)。A.westerdijkiae主要报道其次生代谢产物OchratoxinA污染农产品,水果,和各种食品,如咖啡豆(Alvindiaetal2016),葡萄(Díaz等人。2009),橙子和果汁(Marino等人2009),等。据我们所知,这是中国首例引起赤霉病的赤霉病。
    Rehmannia glutinosa (also known as Chinese foxglove) is a perennial dicotyledonous herb, which plays an important role in traditional Chinese medicine. Its active ingredients have a wide range of pharmacological effects on the blood system, endocrine system, immune system, cardiovascular system, and nervous system (Zhang et al. 2008). In May 2022, leaf blight was observed on 45-day-old R. glutinosa in a seedling nursery in Jiaozuo City (35°01\'44.20″N, 113°05\'30.63″E), Henan Province, China with an approximate disease incidence up to 54% (~1,300 plants). Irregular brown lesion initially appeared on the tips of basal leaves, then progressed to the entire leaf causing leaf drying out (Supple. Fig. 1-A, B, C). The same symptoms appeared successively in the leaves from the base to the top of the plant, which eventually caused the whole plant to die. To identify the pathogen, eight symptomatic leaves were randomly collected from eight individual plants, and cut into small pieces (5 × 5 mm) at the border of lesions. The pieces were surface disinfected in 75% ethanol for 15 s, followed by 1% NaClO for 1 min, rinsed in sterile water three times, and placed on potato dextrose agar (PDA) medium in the dark for 3 days at 25℃. Finally, 12 purified isolates (DHY1-DHY12) were obtained by using single spore method. Leaves of R. glutinosa seedlings were inoculated with conidial suspension (106 conidia/ml), three plants were inoculated per isolate. Controls were treated with sterilized water. All inoculated and control plants were incubated in a greenhouse at 25℃ under 80 ± 10% humidity and a 8-h/16-h dark/light cycle. This experiment was repeated three times. After 5 days, similar symptoms to those of diseased leaves in the seedling nursery appeared on leaves inoculated with DHY4-DHY10, while plants inoculated with DHY1-DHY3, DHY11-DHY12, and the controls remained asymptomatic (Supple. Fig.1-D, E). The same fungi were re-isolated from diseased leaves, fulfilling Koch\'s postulates. The causal agents DHY4 to DHY10, showed similar morphology, which were morphologically identified as Aspergillus sp. (Visagie et al. 2014). Isolate DHY5 was selected for further study. On PDA plates, the colonies were covered with white velutinous mycelia (Supple. Fig.1-F). Conidia were ochre yellow and outwards concentric circles. Vesicles were globose, and about 20.1-26.6 μm in diameter (Supple. Fig.1-G). Conidiophore stipes were smooth walled and hyaline, with conidial heads radiating. The conidia were light yellow to orange, exudate clear to orange droplets. The conidia were (2.53-3.25) μm × (2.58-3.47) μm in diameter (n=50) (Supple. Fig.1-H). For further molecular identification, the ITS and TUB gene sequences were amplified with primer pairs ITS1/ITS4 and BT2a/BT2b (Glass and Donaldson. 1995), respectively. BLASTn searches of the ITS (PP355445) and TUB (PP382788) sequences showed 100% and 98.42% similarity to those of A. westerdijkiae (OP237108 and OP700424), respectively. Phylogenetic analysis based on the concatenated sequences of ITS and TUB confirmed that the fungus was A. westerdijkiae, (Supple. Fig.2). A. westerdijkiae was mainly reported on its secondary metabolite ochratoxin A contamination of agricultural products, fruits, and various food products, such as coffee beans (Alvindia et al 2016), grapes (Díaz et al. 2009), oranges and fruit juice (Marino et al. 2009), etc. To our knowledge, this is the first report of A. westerdijkiae causing leaf blight on R. glutinosa in China.
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  • 文章类型: Journal Article
    双盲,安慰剂对照,随机化,概念验证试验旨在评估VerbasnolTM[地黄叶基提取物(RGLE)]在女性中的疗效和安全性,中度至重度寻常痤疮。
    22名年龄在18至35岁之间的中度至重度痤疮,全球痤疮分级系统(GAGS)评分为19至38的女性被纳入研究,并以1:1的比例随机分配,早餐后口服1粒RGLE胶囊(100mg/天)或安慰剂56天。主要结果是在第56天与安慰剂相比,通过GAGS测量的痤疮严重程度的变化。次要结果是炎性痤疮病变数量的变化,面部皮脂分泌,生活质量,局部疼痛和瘙痒,皮肤皱纹严重程度,和其他皮肤特征,包括辐射,光度,平滑度,纹理,坚定,和水合作用。此外,评估了应答者的百分比以及总体耐受性和疗效.
    在RGLE(n=10)和安慰剂组(n=12)第28天,平均GAGS评分分别降低了21.72%和14.20%,分别,在第56天,两组进一步减少。RGLE组在第56天报告了其他皮肤特征的更好改善。未报告提取物的安全性或耐受性问题。早在补充28天,与安慰剂相比,RGLE减少了女性的痤疮并改善了皮肤质量。
    剂量为100mg/天的RGLE补充剂在56天的剂量给药后,临床上可降低痤疮严重程度,并改善痤疮参与者的皮肤水合作用和生活质量。
    UNASSIGNED: A double-blind, placebo-controlled, randomized, proof-of-concept trial aimed to evaluate the efficacy and safety of VerbasnolTM [Rehmannia glutinosa Libosch leaf-based extract (RGLE)] in females, with moderate to severe acne vulgaris.
    UNASSIGNED: Twenty-two females aged 18 to 35 years having moderate to severe acne with Global Acne Grading System (GAGS) scores of 19 to 38 were included in the study and were randomized in a 1:1 ratio to receive either one capsule (100 mg/day) of RGLE or placebo orally after breakfast for 56 days. The primary outcome was a change in acne severity measured by the GAGS compared to the placebo on day 56. The secondary outcomes were changes in the number of inflammatory acne lesions, facial sebum secretion, quality of life, local pain and itching, skin wrinkle severity, and other skin characteristics, including radiance, luminosity, smoothness, texture, firmness, and hydration. Additionally, the percentage of responders and global tolerability and efficacy were evaluated.
    UNASSIGNED: The mean GAGS score was reduced by 21.72% and 14.20% on day 28 in RGLE (n=10) and placebo groups (n=12), respectively, which further reduced in both groups on day 56. The RGLE group reported better improvement in other skin characteristics on day 56. No safety or tolerability concerns were reported for the extract. RGLE reduced acne and improved the skin quality in females compared to placebo as early as 28 days of supplementation.
    UNASSIGNED: RGLE supplementation at a dose of 100 mg/day has provided a clinically relevant decrease in acne severity and improved the skin hydration and quality of life of the participants with acne after 56 days of dose administration.
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  • 文章类型: Journal Article
    2型糖尿病(T2DM)对全球人口健康构成巨大威胁,需要探索更多的治疗药物。在这项研究中,我们研究了地黄中有效成分catalpol降低糖尿病血糖的机制。
    通过SD大鼠腹腔注射链脲佐菌素构建T2DM模型,随机分为糖尿病模型组,吡格列酮组,地黄组,catalol高剂量组,枸橼酸低剂量组及正常对照组。干预持续28天,和体重的变化,空腹血糖,观察胰岛素和血脂水平。
    在所有的药物中,吡格列酮具有最明显的降血糖作用,低剂量catalpol组治疗2周后开始下降,而高剂量catalpol组没有降血糖作用。其中,地黄能够增加血清甘油三酯水平,吡格列酮可有效降低大鼠总胆固醇水平。低剂量的梓醇降低了低密度脂蛋白胆固醇(LDL)的浓度,而高剂量的梓醇增加了LDL的浓度。
    作为地黄的活性成分,catalpol在糖尿病治疗中具有降低血糖和改善血脂的潜力,其作用可能是通过调节腺苷5'-单磷酸(AMP)激活的蛋白激酶(AMPK)信号通路来实现的,为糖尿病治疗新方法的发展提供了新的思路。
    UNASSIGNED: Type 2 diabetes mellitus (T2DM) poses a huge threat to population health globally, and more drugs need to be explored for treatment. In this study, we investigated the mechanism of active ingredient catalpol in Rehmannia glutinosa on reduces blood glucose in diabetic.
    UNASSIGNED: The T2DM model was constructed by intraperitoneal injection of streptozotocin into Sprague-Dawley (SD) rats, which were randomly grouped into diabetes model group, pioglitazone group, Rehmannia glutinosa group, catalpol high-dose group, catalpol low-dose group and normal control group.The intervention was continued for 28 d, and changes in body weight, fasting blood glucose, insulin and lipid levels were observed.
    UNASSIGNED: Of all the drugs, pioglitazone had the most pronounced hypoglycemic effect, which began to decline after 2 weeks of treatment in the low-dose catalpol group and had no hypoglycemic effect in the high-dose catalpol group. Among them, Rehmannia glutinosa was able to increase serum triglyceride level, and pioglitazone effectively reduced total cholesterol level in rats. The low dose of catalpol decreased the concentration of low-density lipoprotein cholesterol (LDL), while the high dose of catalpol increased the concentration of LDL.
    UNASSIGNED: As an active ingredient in Rehmannia glutinosa, catalpol has the potential to lower blood glucose and improve blood lipids in diabetes treatment, and its action may be achieved by regulating the adenosine 5\'-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway, which provides a new idea for the development of new diabetes therapeutic approaches.
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  • 文章类型: Journal Article
    本研究旨在确定三种抗炎材料的最佳组合[即,CervusnipponTemminck(CT),当归gigasNakai(AN),和地黄(RG)]具有最强的抗炎潜力。通过评估一氧化氮(NO),在LPS刺激的RAW264.7细胞中测试了三种材料的18种组合。来自体外研究的最佳组合给予LPS处理的C57BL/6J小鼠5天。随后,通过生物信息学分析和验证对血浆代谢物进行了分析.作为结果,2、20和50µg/mL的CT,AN,和RG(TM)是抑制炎症最有效的组合。在老鼠身上,TM减轻肝脏炎症标志物。同样,代谢组学表明TM可能抑制NF-κB信号通路,从而减轻肝脏炎症。TM还降低全身性和肝脏促炎细胞因子。总的来说,我们发现了缓解炎症的TM的最佳组合;因此,对安全性的进一步研究,机制,和临床模型对于人类应用是有保证的。
    在线版本包含补充材料,可在10.1007/s10068-023-01476-x获得。
    This study aimed to determine the optimal combination of three anti-inflammatory materials [i.e., Cervus nippon Temminck (CT), Angelica gigas Nakai (AN), and Rehmannia glutinosa (RG)] for the strongest anti-inflammatory potential. Eighteen combinations of the three materials were tested in LPS-stimulated RAW264.7 cells via assessing nitric oxide (NO). The best combination from in vitro studies was administered to LPS-treated C57BL/6J mice for five days. Subsequently, plasma metabolites were profiled by bioinformatics analyses and validations. As results, 2, 20, and 50 µg/mL of CT, AN, and RG (TM) were the most effective combination suppressing inflammation. In mice, TM mitigated hepatic inflammatory markers. Similarly, the metabolomics indicated that TM may suppress NF-κB signaling pathway, thereby alleviating hepatic inflammation. TM also decreased systemic and hepatic pro-inflammatory cytokines. Collectively, we found the optimal combination of TM for mitigating inflammation; thus further studies on safety, mechanisms, and clinical models are warranted for human applications.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s10068-023-01476-x.
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  • 文章类型: Journal Article
    五种未描述的嗜乙菌环型倍半萜,瑞亲生三醇E-I(1-5),从地黄的新鲜根中分离出7种已知化合物(6-12)。通过广泛的光谱数据分析对其结构进行了表征,并通过比较其计算的电子圆二色性(ECD)光谱和实验ECD光谱来确定其绝对构型。通过MTT方法评价了所有化合物的体外抗肺纤维化活性。化合物2、8、10和12表现出优异的抗肺纤维化活性。此外,化合物2可以降低TGF-β1诱导的BEAS-2B细胞的ROS水平和凋亡。
    Five undescribed eremophilane-type sesquiterpenes, remophilanetriols E-I (1-5), along with seven known compounds (6-12) were isolated from the fresh roots of Rehmannia glutinosa. Their structures were characterized by extensive spectroscopic data analysis and their absolute configurations were determined by comparing their calculated electronic circular dichroism (ECD) spectra and experimental ECD spectra. The anti-pulmonary fibrosis activities of all compounds were evaluated in vitro by MTT methods, and compounds 2, 8, 10, and 12 exhibited excellent anti-pulmonary fibrosis activities. In addition, compound 2 can reduce the levels of ROS and apoptosis in TGF-β1-induced BEAS-2B cells.
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  • 文章类型: Journal Article
    地黄,玄参科的一员,自古以来就被广泛应用于中药中。谷红的主要生物活性成分是梓醇。然而,catalpol的生物发生,尤其是它的下游途径,尚不清楚。为了确定参与catalol生物合成的候选基因,转录组是使用三个品种的幼叶从R.glutinosa构建的,北京一号3,怀丰,和JinNo.9,以及JinNo.的块根和不定根。9个品种。因此,产生了71,142个具有功能注释的单基因。对R.glutinosa转录组的比较分析确定了超过200种单基因的13种酶,这些酶可能参与catalpol形成的下游步骤。包括9个编码UGTs的基因,13用于醛脱氢酶,70用于氧化还原酶,44对于CYP450,22用于脱水酶,30用于脱羧酶,19用于羟化酶,10为环氧酶。此外,两个编码香叶醇合酶(RgGES)的新基因,这是人们在catalpol生产中的第一个酶,是从r.glutinosa克隆的。RgGESs的纯化的重组蛋白有效地将miR转化为香叶醇。这项研究首次发现了在catalpol生物合成中编码上述定制酶的推定基因,并在功能上表征了该途径中的酶编码基因。该结果丰富了基因工程的遗传资源,可用于设计过氧化氢和环烯醚萜类的生物合成途径。
    Rehmannia glutinosa, a member of the Scrophulariaceae family, has been widely used in traditional Chinese medicine since ancient times. The main bioactive component of R. glutinosa is catalpol. However, the biogenesis of catalpol, especially its downstream pathway, remains unclear. To identify candidate genes involved in the biosynthesis of catalpol, transcriptomes were constructed from R. glutinosa using the young leaves of three cultivars, Beijing No. 3, Huaifeng, and Jin No. 9, as well as the tuberous roots and adventitious roots of the Jin No. 9 cultivar. As a result, 71,142 unigenes with functional annotations were generated. A comparative analysis of the R. glutinosa transcriptomes identified over 200 unigenes of 13 enzymes potentially involved in the downstream steps of catalpol formation, including 9 genes encoding UGTs, 13 for aldehyde dehydrogenases, 70 for oxidoreductases, 44 for CYP450s, 22 for dehydratases, 30 for decarboxylases, 19 for hydroxylases, and 10 for epoxidases. Moreover, two novel genes encoding geraniol synthase (RgGES), which is the first committed enzyme in catalpol production, were cloned from R. glutinosa. The purified recombinant proteins of RgGESs effectively converted GPP to geraniol. This study is the first to discover putative genes coding the tailoring enzymes mentioned above in catalpol biosynthesis, and functionally characterize the enzyme-coding gene in this pathway in R. glutinosa. The results enrich genetic resources for engineering the biosynthetic pathway of catalpol and iridoids.
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  • 文章类型: Journal Article
    中药(TCM)地黄(RR)是指玄参科植物地黄的新鲜或干燥的块根。作为一种传统的中草药(CHM),它具有多种效果,包括镇痛,镇静,抗炎,抗氧化,抗肿瘤,免疫调节,心脑血管调节,和神经损伤修复,在临床上得到了广泛的应用。近年来,科学家已经广泛研究了RR的活性成分和药理作用。活性成分主要包括环烯醚萜苷类(如catalpol和aucuboside),类苯丙素糖苷(如乙酰糖苷),其他糖类,和不饱和脂肪酸。此外,与RR相关的中成药(CPM)和中药汤剂也成为中医从业者的主要研究课题之一,其中包括地黄,百合鳞茎地黄汤,和四物汤。本文回顾了近年来关于RR的文献;总结了其化学成分的研究,药理作用,和临床应用;并分析了当前研究的进展和局限性,为RR的进一步探索和发展提供参考。
    The traditional Chinese medicine (TCM) Rehmanniae Radix (RR) refers to the fresh or dried root tuber of the plant Rehmannia glutinosa Libosch of the family Scrophulariaceae. As a traditional Chinese herbal medicine (CHM), it possesses multiple effects, including analgesia, sedation, anti-inflammation, antioxidation, anti-tumor, immunomodulation, cardiovascular and cerebrovascular regulation, and nerve damage repair, and it has been widely used in clinical practice. In recent years, scientists have extensively studied the active components and pharmacological effects of RR. Active ingredients mainly include iridoid glycosides (such as catalpol and aucuboside), phenylpropanoid glycosides (such as acteoside), other saccharides, and unsaturated fatty acids. In addition, the Chinese patent medicine (CPM) and Chinese decoction related to RR have also become major research subjects for TCM practitioners; one example is the Bolus of Six Drugs, which includes Rehmannia, Lily Bulb and Rehmannia Decoction, and Siwu Decoction. This article reviews recent literature on RR; summarizes the studies on its chemical constituents, pharmacological effects, and clinical applications; and analyzes the progress and limitations of current investigations to provide reference for further exploration and development of RR.
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  • 文章类型: Journal Article
    地黄在中医治疗糖尿病中被广泛用于控制血糖。在本研究中,三个新化合物,包括一个环烯醚萜类地黄A(1)和两个紫罗兰地黄B-C(7-8),连同14种已知化合物(2-6和9-17),是从r.glutinosa的根中分离出来的。通过物理化学常数和光谱分析(1D,2D-NMR和MS)。体外测试了1-17对α-葡萄糖苷酶活性的影响。化合物9、10和11(IC50:5.0、3.1和6.3mM)显示出相对于阿卡波糖抑制α-葡糖苷酶的中等活性(IC50=3.0mM)。该研究结果为了解赤霉病的降血糖作用和α-葡萄糖苷酶抑制剂药物的开发提供了一些新的见解。
    Rehmannia glutinosa was extensively used to control blood sugar in diabetes treatment in tradition Chinese medicine. In the present study, three new compounds, including an iridoid rehmannia A (1) and two ionone rehmannias B-C (7-8), together with fourteen known compounds (2-6 and 9-17), were isolated from the roots of R. glutinosa. The structures of these compounds were determined by physicochemical constants and spectral analysis (1D, 2D-NMR and MS). The effect of 1-17 on α-glucosidase activity was tested in vitro. Compounds 9, 10, and 11 (IC50: 5.0, 3.1, and 6.3 mM) showed moderate activity to suppress α-glucosidase relative to acarbose (IC50 = 3.0 mM). The findings provided some new insights to understand the hypoglycemic effect of R. glutinosa and the development towards the α-glucosidase inhibitor drugs.
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  • 文章类型: Journal Article
    我们报告了成功开发的磁性分子印迹聚合物(MMIP),用于分离和检测地黄提取物和生物样品中的梓醇。Catalpol被用作MMIP的模板分子,使用反向预测方法和差示紫外-可见光谱合成,以优化合成条件。使用包括扫描电子显微镜在内的技术对所得的MMIP进行了表征,透射电子显微镜,傅里叶变换红外光谱,和振动样品磁力测定,这证实了他们优秀的核壳结构和简单的快速磁分离。进行了静态和动态吸附实验以评估灵敏度,重复性,和MMIP的吸附能力。发现吸附容量很高(59.09µg/mg)。MMIP还表现出对catalol类似物的强鉴定特异性(IF=2.41)。对磁性固相萃取(MSPE)进行了优化,以建立基于MMIP的MSPE-HPLC方法。在理想条件下,新方法在0.1-1.0mg/mL和0.03-0.18mg/mL范围内显示出优异的测定系数(R2>0.99)。分别,检出限为5.4ng/mL和1.7ng/mL。这项研究的新颖之处在于使用磁性载体制备磁性表面MMIP。这些MMIP表现出优异的磁性能,并具有对catalpol的特定吸附能力,实现快速分离。它们具有低成本等优点,高特异性,化学稳定性,物理鲁棒性,和可回收性(97.09%-106.53%,91.32%-100.99%)。将这些聚合物应用于测定赤霉酸提取物和生物样品中梓醇含量的预处理方法中,提高了梓醇的提取效率和检测准确性。它消除了Catalpol测定中其他成分的干扰和影响,同时具有很高的灵敏度,良好的富集效果,优秀的可重用性,和高回收率。
    We report on the successful development of a magnetic molecularly imprinted polymer (MMIP) for the separation and detection of catalpol in Rehmannia glutinosa extract and biological samples. Catalpol was used as the template molecule for the MMIPs, which were synthesized using the reverse prediction approach and differential UV-visible spectra to optimize the synthesis conditions. The resulting MMIPs were characterized using techniques including scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and vibrating sample magnetometry, which confirmed their excellent nuclear shell construction and simple rapid magnetic separation. Static and dynamic adsorption experiments were performed to assess the sensitivity, repeatability, and adsorption capacity of the MMIPs. Adsorption capacity was found to be high (59.09 µg/mg). The MMIPs also exhibited strong identification specificity for catalpol analogs (IF = 2.41). Magnetic solid phase extraction (MSPE) was optimized to establish an MSPE-HPLC approach based on the MMIPs. Under ideal conditions, the new method showed excellent determination coefficient (R2 > 0.99) in the ranges of 0.1-1.0 mg/mL and 0.03-0.18 mg/mL for R. glutinosa extract and biological material, respectively, with limits of detection of 5.4 ng/mL and 1.7 ng/mL. The novelty of this research lies in the preparation of magnetic surface MMIPs using a magnetic carrier. These MMIPs exhibit excellent magnetic properties and possess specific adsorption capability for catalpol, enabling rapid separation. They offer advantages such as low cost, high specificity, chemical stability, physical robustness, and recyclability (97.09%-106.53%, 91.32%-100.99%). The application of these polymers in the pretreatment method for determining the catalpol content in R. glutinosa extract and biological samples enhances the extraction efficiency and detection accuracy for catalpol. It eliminates interference and influences from other components in catalpol determination while exhibiting high sensitivity, good enrichment effects, excellent reusability, and a high recovery rate.
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