Molecular authentication of the traditional Chinese medicinal plant Euphorbia pekinensis

The ITS regions of Euphorbia pekinensis and six other Euphorbia species used as adulterants of E. pekinensis were sequenced to differentiate them. The sequences are identical among the individuals in the seven species studies. Diversity in DNA sequences among various species was found ranging from 8.3% to 43.8% in ITS1 and 7.6% to 36.6% in ITS2 region. Furthermore, based on the divergent ITS regions, species-specific primers, JDJp 1 and JDJp 2, were designed in the polymorphic regions of E. pekinensis to distinguish it from adulterants. These ITS-derived primers amplified a 281-bp-specific DNA fragment from E. pekinensis. No amplified product was observed using DNA of six adulterants.     

Molecular authentication of the traditional dai medicinal plant Croton caudatus

Croton caudatus is an important species in Dai folk medicine. Some other Croton species, in particular, C. kongensis, C. cascarilloides, C. crassifolius, C. lachnocarpus and C. olivaceus are often marketed as C. caudatus, and thus, the therapeutic effects of C. caudatus are not achieved. The respective morphological features of the plants are similar, and they are not easy to distinguish morphologically from each other. In an attempt to find a method for discriminating among these species, we compared their nuclear ribosomal DNA internal transcribed spacer (ITS) region. Species-specific probes were derived from the ITS region of these species for species identification. This method provides effective and accurate identification of C. caudatus.     

RP-HPLC测定藏药川西獐芽菜中的龙胆苦苷

目的 建立了测定不同地区川西獐芽菜中的龙胆苦苷.方法 采用RP-HPLC法,Kromasil C18色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇-0.04%磷酸水溶液(23:77),流速1.0 ml · min-1,检测波长238 nm,柱温35℃.结果 龙胆苦苷的线性范围为0.026～1.048 μg ( r＝0.9996),平均回收率为99.6%,RSD＝1.54%.结论 所建方法简便、准确、重复性好,适用于川西獐芽菜及其他药用植物中的龙胆苦苷.     

Application of real-time scorpion PCR for authentication and quantification of the traditional Chinese medicinal plant Drynaria fortunei

DNA sequence analysis of the trnl-trnF spacer region and real-time scorpion polymerase chain reaction were exploited for their applications in the differentiation and quantification of the traditional Chinese medicinal plants Drynaria fortunei from five related adulterants: D. mollis, D. quercifolia, D. rigidula, D. sparsisora, and Pseudodrynaria coronans. The data demonstrated that variations in the trnl-trnF spacer regions were very low at the intraspecific level but extremely high at the interspecific level, meaning that they could be easily distinguished at the DNA level. The sequence difference allowed effective and reliable differentiation of D. fortunei from adulterants by real-time scorpion PCR. Furthermore, a quantification methodology was carried out to quantify D. fortunei.     

HPLC法测定川西獐牙菜中獐牙菜苷的含量

目的建立川西獐牙菜所含獐牙菜苷的HPLC定量测定方法。方法采用HPLC法。色谱柱:C18柱(200mm×4.6mm,5μm)柱;流动相:乙腈-0.1%磷酸(10∶90);流速为1.0mL·min-1;检测波长:237nm;柱温:30℃。结果獐牙菜苷在0.24～1.2μg范围内线性关系良好,平均回收率为102.6%,RSD为2.2%。结论所建立的检测方法可快速有效的测定獐牙菜苷。     

A DNA microarray for the authentication of toxic traditional Chinese medicinal plants

A silicon-based DNA microarray was designed and fabricated for the identification of toxic traditional Chinese medicinal plants. Species-specific oligonucleotide probes were derived from the 5S ribosomal RNA gene of Aconitum carmichaeli, A. kusnezoffi, Alocasia macrorrhiza, Croton tiglium, Datura inoxia, D. metel, D. tatula, Dysosma pleiantha, Dy. versipellis, Euphorbia kansui, Hyoscyamus niger, Pinellia cordata, P. pedatisecta, P. ternata, Rhododendron molle, Strychnos nux-vomica, Typhonium divaricatum and T. giganteum and the leucine transfer RNA gene of Aconitum pendulum and Stellera chamaejasme. The probes were immobilized via dithiol linkage on a silicon chip. Genomic target sequences were amplified and fluorescently labeled by asymmetric polymerase chain reaction. Multiple toxic plant species were identified by parallel genotyping. Chip-based authentication of medicinal plants may be useful as inexpensive and rapid tool for quality control and safety monitoring of herbal pharmaceuticals and neutraceuticals.     

藏茵陈对重症胰腺炎大鼠炎症因子表达的影响

目的研究藏茵陈对重症胰腺炎大鼠的保护作用。方法将大鼠分为假手术组、模型组及藏茵陈高、中、低剂量组,灌胃给予药物,连续给药14 d后造模。采用生化法测定血清淀粉酶(AMS)的活力;用酶联免疫吸附法测定血清中肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、白介素-6(IL-6)和单核细胞趋化蛋白-1(MCP-1)的蛋白表达;用酶联免疫吸附法测定胰腺中IL-6和MCP-1的蛋白表达;用实时荧光定量PCR测定胰腺中MCP-1和IL-6的基因表达;用HE染色观察大鼠胰腺病理学组织。结果藏茵陈高剂量组AMS活力及血清中TNF-α、IL-6和MCP-1的蛋白表达明显降低,IL-10的蛋白表达升高(P<0.05)。藏茵陈可显著降低胰腺中MCP-1的蛋白及基因表达(P<0.05),对IL-6的蛋白和基因表达影响差异无统计学意义(P>0.05)。结论藏茵陈对重症胰腺炎大鼠具有一定的保护作用,其机制可能是降低了胰腺中MCP-1的蛋白和基因表达。     

川西獐牙菜的化学成分、药理作用和临床应用研究进展

川西獐牙菜是我国传统藏药"藏茵陈"原植物之一,含有呫吨酮、环烯醚萜苷、三萜等化学成分,具有保肝、抑菌等药理作用。其临床应用主要为各型肝炎的治疗,已被开发为多种剂型的成药,该植物其他方面的应用还有待于进一步研究。对该植物的化学成分、药理作用和临床应用研究进展做一综述,以期为开发和利用川西獐牙菜提供参考。     

Authentication of the traditional medicinal plant Eleutherococcus senticosus by DNA and chemical analyses

Shigoka (SGK), the rhizome of Eleutherococcus senticosus, is a traditional medicine used as a tonic in northeastern Asia and far eastern Russia. We analyzed the nuclear ribosomal DNA internal transcribed spacer (ITS) sequence of the medicine available on the Japanese and Chinese markets and found that at least 3 species were used as the source plant of the commercial SGKs and that only 70% of all samples was made from the correct species. Furthermore, we performed the quantitative determination of 3 marker compounds, eleutheroside B (EB), syringaresinol diglucoside (Syr), and isofraxidin (Iso) by ultraperformance liquid chromatography (UPLC)/mass spectrometry (MS). We found that EB and Iso are specific to the correct source plant of SGK. Of them, EB is thought to be the best marker compound for quality assurance of the SGK from the viewpoint of its pharmacological activity.     

Genome-based approaches to the authentication of medicinal plants

Medicinal plants are the source of a large number of essential drugs in Western medicine and are the basis of herbal medicine, which is not only the primary source of health care for most of the world's population living in developing countries but also enjoys growing popularity in developed countries. The increased demand for botanical products is met by an expanding industry and accompanied by calls for assurance of quality, efficacy and safety. Plants used as drugs, dietary supplements and herbal medicines are identified at the species level. Unequivocal identification is a critical step at the beginning of an extensive process of quality assurance and is of importance for the characterization of the genetic diversity, phylogeny and phylogeography as well as the protection of endangered species. DNA-based methods have been developed for the identification of medicinal plants. Nuclear and chloroplast DNA is amplified by the polymerase chain reaction and the reaction products are analyzed by gel electrophoresis, sequencing, or hybridization with species-specific probes. Genomic fingerprinting can differentiate between individuals, species and populations and is useful for the detection of the homogeneity of the samples and presence of adulterants. Although sequences from single chloroplast or nuclear genes have been useful for differentiation of species, phylogenetic studies often require consideration of DNA sequence data from more than one gene or genomic region. Phytochemical and genetic data are correlated but only the latter normally allow for differentiation at the species level. The generation of molecular "barcodes" of medicinal plants will be worth the concerted effort of the medicinal plant research community and contribute to the ongoing effort of defining barcodes for every species on earth.     

Molecular authentication of the ethnomedicinal plant Sabia parviflora and its adulterants by DNA barcoding technique

Sabia parviflora Wall. ex Roxb. is a traditional herb widely used by Chinese people, especially by the Buyi ethnic group which resides in Guizhou and Yunnan provinces. According to the Chinese Ethnic Pharmacopeia, the species is commonly used for soothing the liver and for the treatment of icteric hepatitis, hemostasis, and inflammation. However, due to the similar morphological characters of Sabia species and higher market demands, there are many substitutes and adulterants of S. parviflora. In this study, the differential identification of 6 Sabia species and 7 adulterants were investigated through DNA sequence analysis of three candidate DNA barcodes (trnH-psbA, rbcL-α, matK). Based on sequence alignments, we concluded that not only the trnH-psbA spacer sequence can distinguish S. parviflora from other Sabia species, but the matK + rbcL-α sequences also can differentiate it from the substitutes and adulterants. The classification tree of all samples based on rbcL-α sequences indicated that the rbcL region can identify samples into a family/genus level. Our results suggest that the three candidate barcodes can be used for the identification of S. parviflora and to distinguish it from common substitutes or adulterants.     

Hepatoprotective and anti-inflammatory effects of a traditional medicinal plant of Chile, Peumus boldus

Dried hydro-alcoholic extract of Peumus boldus (Monimiaceae) has been evaluated for hepatoprotective, choleretic and anti-inflammatory effects in mice and rats, in order to validate or to invalidate traditional therapeutic indications. This extract exerted a significant hepatoprotection of tert-butyl hydroperoxide-induced hepatotoxicity in isolated rat hepatocytes (in vitro technique) by reducing the lipid peroxidation and the enzymatic leakage of LDH; this in vitro efficacy was reinforced by a significant hepatoprotection on CCl4-induced hepatotoxicity in mice (in vivo technique), the plant extract reducing the enzymatic leakage of ALAT. Boldine, the main alkaloid of P. boldus appears to be implicated in this hepatoprotective activity. Choleretic effects, often mentioned in traditional indications, have not been confirmed in rats. Finally, significant and dose-dependent anti-inflammatory effects were obtained on an acute inflammatory process (carrageenan-induced edema test in rats). Boldine does not appear to be involved in such properties.     

Neurotoxicity of Coscinium fenestratum stem, a medicinal plant used in traditional medicine.

Coscinium fenestratum is a common medicinal plant widely used in the Indochina region, but scientific data on its safety is very limited. This study aimed to observe the effect of this plant on neurotoxicity and neurobehavior. Oral administration of plant alcoholic extract at dosages of 5, 10 and 20 mg/kgBW for 14 days increased the rats body weight and decreased the neuron density in the cerebral cortex, hippocampus and striatum. The plant extract significantly increased stereotyped behavior in licking but did not cause anxiolytic activity, anti-depression, sensory motor co-ordination impairment and ataxia. It is concluded that the plant possesses neurotoxicity and is able to induce neurobehavioral changes in rats. Therefore, the application of this plant as either drug or supplementary food should be reconsidered.     

川西獐牙菜乙醇提取物对咖啡因依赖小鼠戒断反应的抑制及其镇痛作用

目的 观察川西獐牙菜75％乙醇提取物(SME)的镇痛作用和对咖啡因依赖小鼠戒断反应的抑制作用.方法 将小鼠随机分为4组,分别以生理盐水、咖啡因和SME不同组合处理小鼠,每天两次,连续10 d;第10天初次处理后,注射纳洛酮引起戒断综合症,观察SME对戒断反应的抑制作用;在实验的第1、3、6、9、10天,用热板实验观察SME的镇痛作用;将小鼠随机分为空白组、模型组、阳性(阿司匹林)组和SME不同剂量组,分别给予相应药物,qd,连续8d,通过扭体实验和血清生化指标的检测,进一步验证SME的镇痛作用.结果 SME可抑制咖啡因引起的镇痛高峰期,减少小鼠在戒断反应中的跳跃次数;扭体实验中,给予小鼠不同剂量SME后,扭体次数和给药剂量之间呈负相关,并明显降低血清中前列素E2和一氧化氮的含量.结论 SME有显著的镇痛作用和潜在的抑制戒断反应的作用.     

Evaluation of the antioxidants activities of four Slovene medicinal plant species by traditional and novel biosensory assays

We investigated the antioxidant activity of methanolic and water extracts of Slovene accessions of four medicinal plant species (Salvia officinalis, Achillea millefolium, Origanum vulgare subsp. vulgare and Gentiana lutea). Their free radical-scavenging activity against the DPPH· free radical was studied with a spectrophotometric assay, while their biological activity with the help of a laboratory-made biosensor based on immobilized fibroblast cells (assay duration: 3 min). The observed antioxidant activity of the extracts from the four investigated medicinal plant species was dependent on both the solvent used for extraction and the assay method (conventional or biosensor-based). Independently from the assay method and the solvent used for extraction, the lowest scavenging activity was observed in root extracts of G. lutea. Treatment of the immobilized cells with the plant extracts resulted in an increase of the cell membrane potential (membrane hyperpolarization), possibly due to the reduction of membrane damage due to oxidation. The novel cell biosensor could be utilized as a rapid, high throughput tool for screening the antioxidant properties of plant-derived compounds.     

Hepatoprotective properties of Commiphora opobalsamum ("Balessan"), a traditional medicinal plant of Saudi Arabia

The hepatoprotective activity of an ethanolic extract of Commiphora opobalsamum ("Balessan") was investigated in rats by inducing hepatotoxicity with carbon tetrachloride:liquid paraffin (1:1). This extract has been shown to possess significant protective effect by lowering serum transaminase levels (serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase), alkaline phosphatase and bilirubin. Pretreatment with an extract of Balessan prevented the prolongation of the barbiturate sleeping time associated with carbon tetrachloride-induced liver damage in mice. On the other hand, CCl4-induced low-level nonprotein sulfhydryl concentration in the liver was replenished by the Balessan extract. These data suggest that the plant C. opobalsamum may act as an antioxidant agent and may have a hepatoprotective effect.     

Antihypertensive actions of methylripariochromene A from Orthosiphon aristatus, an Indonesian traditional medicinal plant.

Methylripariochromene A (MRC) was isolated from the leaves of Orthosiphon aristatus (Lamiaceae) and subjected to the examination of several pharmacological actions related to antihypertensive activity. The following four findings were revealed from the present study: 1) MRC caused a continuous decrease in systolic blood pressure and a decrease in heart rate after subcutaneous administration in conscious male SHRSP, 2) MRC exhibited the concentration-dependent suppression of contractions induced by high K+, l-phenylephrine or prostaglandin F2alpha in endothelium-denuded rat thoracic aorta, 3) MRC showed a marked suppression of contractile force without a significant reduction in the beating rate in isolated bilateral guinea pig atria, and 4) MRC increased urinary volume and the excretion of Na+, K+ and Cl- for 3 h after oral administration with a load of saline in fasted rats. These findings indicate that MRC possesses some actions related to a decrease in blood pressure, i.e. vasodilating action, a decrease in cardiac output and diuretic action. Furthermore, it is presumed that the traditional use of this plant in the therapy of hypertension may be partially supported by these actions with MRC.