凤尾草抗糖尿病活性成分的研究

目的研究凤尾草抗糖尿病的活性成分。方法凤尾草全草用75%乙醇提取,利用硅胶、凝胶等柱色谱及制备液相色谱进行分离,并根据理化性质和有机波谱技术鉴定了化合物结构,进一步通过测定黄酮苷类化合物对胰岛素抵抗的HepG2细胞葡萄糖消耗量以及对AMPK和ACC磷酸化水平的影响来评价其抗糖尿病活性。结果从凤尾草中分离鉴定了9个化合物,分别为:芹菜素-4'-O-α-L-鼠李糖苷(1),芹菜素-7-O-β-D-葡萄糖-4'-O-α-L-鼠李糖苷(2),柚皮素-7-O-β-D-葡萄糖苷(3),木犀草素(4),2β,5α-二羟基-(-)-16-贝壳杉烯(5),pterosin B(6),24R-ethylcholest-4-en-3-one(7),cassipourol(8),cyclolaudenol(9),此外化合物(2)能够显著提高胰岛素抵抗HepG2细胞葡萄糖消耗及AMPK和ACC磷酸化水平。结论化合物(7)～(9)为首次从该植物中分离得到,化合物(2)具有较强的抗糖尿病活性。     

HepG2细胞胰岛素抵抗模型的建立及在筛选两色金鸡菊活性化合物中的应用

目的:体外建立人肝癌细胞(HepG2)胰岛素抵抗模型,并初步筛选两色金鸡菊中可有效改善胰岛素抵抗的萃取部位及活性化合物。方法:用不同浓度的胰岛素、棕榈酸或葡萄糖分别对HepG2细胞进行不同时间的诱导,通过MTT法对细胞活性进行评价及葡萄糖氧化酶法对HepG2细胞葡萄糖消耗量测定,明确建立稳定的HepG2胰岛素抵抗模型的试剂诱导浓度及诱导时间。模型建立后,应用不同浓度的两色金鸡菊粗提物、各萃取部位及七个单体化合物奥卡宁(Okanin,2)、3',4',8-三羟基黄酮-7-O-β-D-吡喃葡萄糖苷(3',4',8-trihydroxyflavone-7-O-β-D-glucopyranoside,3)、槲皮素(Quercetin,4)、黄诺马苷(Flavanomarein,5)、马里苷(Marein,6)、豆甾醇(Stigmasterol,13)、紫铆花素(Butein,16)预处理细胞24 h,再用55 mmol·L-1葡萄糖刺激24h,用葡萄糖氧化酶法或2-NBDG荧光标记葡萄糖法分别观察不同浓度的上述成分对胰岛素抵抗模型HepG2细胞糖消耗或糖吸收的影响。MTT法对各组细胞活性进行评价。结果:HepG2细胞在10-7mol·L-1浓度的胰岛素中作用24 h,或在55 mmol·L-1的葡萄糖中刺激24 h,细胞葡萄糖消耗量明显减少(P0.01)且稳定性良好,并且对细胞存活率未造成影响,可作为胰岛素抵抗模型;而棕榈酸组虽然糖消耗显著下降但细胞存活率也显著降低。两色金鸡菊总提物、乙酸乙酯部位、正丁醇部位、水部位及2号、3号、5号和16号化合物均有改善细胞胰岛素抵抗作用,其中5号黄诺马苷和6号马里苷效果较好。结论:黄诺马苷和马里苷可能是两色金鸡菊中改善胰岛素抵抗的主要活性成分。     

金鸡脚化学成分及其抗氧化活性

目的:研究金鸡脚Phymatopteris hastate的化学成分及其抗氧化活性。方法:综合运用硅胶柱色谱、凝胶柱色谱、HPLC制备色谱等多种色谱法分离纯化金鸡脚中的化学成分;采用NMR等波谱学方法鉴定化合物结构;进一步通过DPPH法评价各化合物的抗氧化活性。结果:从金鸡脚乙酸乙酯层和正丁醇层中分离得到14个化合物,分别为4-O-β-D-吡喃葡萄糖基咖啡酸乙酯(1),山柰酚-7-O-α-L-吡喃鼠李糖苷(2),山柰酚-3,7-O-α-L-二吡喃鼠李糖苷(3),山柰酚-3-O-α-L-呋喃阿拉伯糖-7-O-α-L-吡喃鼠李糖苷(4),核桃苷(5),柚皮苷(6),柚皮素-7-O-β-D-吡喃葡萄糖苷(7),反式咖啡酸(8),反式咖啡酸-4-O-β-D-吡喃葡萄糖苷(9),4-O-β-D-吡喃葡萄糖基反式肉桂酸(10),2-O-β-D-吡喃葡萄糖基反式肉桂酸(11),2-O-β-D-吡喃葡萄糖基顺式肉桂酸(12),绿原酸乙酯(13),原儿茶酸(14)。抗氧化实验结果表明:化合物1～9,13,14显示了明确的抗氧化活性。结论:所有化合物均为首次从该属中分离得到;其中化合物1未见文献报道,推测可能为化合物9与乙醇反应后的人工产物;化合物1～9,13,14显示了明确的抗氧化活性。     

茺蔚子萜类化学成分及其抗炎活性研究

目的 研究茺蔚子中的化学成分及其抗炎活性。方法 采用大孔吸附树脂、硅胶柱色谱以及半制备HPLC等多种色谱方法对其化学成分进行分离纯化，结合化合物的理化性质及NMR和MS等波谱数据鉴定化合物结构。通过测定化合物对脂多糖(lipopolysaccharide, LPS)诱导RAW 264.7巨噬细胞释放一氧化氮(NO)的抑制能力来评价化合物的抗炎活性。结果 从茺蔚子正丁醇提取部位中共分离得到4个萜类化合物，含1个新化合物，结构分别鉴定为(-)-(5R,6S,8R)-5,6-二羟基-二氢猕猴桃内酯(1)、蜜柑苷A(2)、淫羊藿次苷C3(3)、益母草宁素K(4)。活性筛选结果表明，化合物4对LPS诱导RAW264.7细胞释放NO具有明显的抑制作用。结论 化合物1是新化合物，化合物3是首次在益母草属植物中分离得到，化合物2和4是首次从茺蔚子中分离得到。化合物4可以显著抑制LPS诱导的RAW264.7细胞的NO释放，具有潜在的抗炎作用。     

马齿苋抗糖尿病活性成分的研究

目的研究马齿苋抗糖尿病的活性成分。方法马齿苋全草用75%乙醇提取,利用硅胶、凝胶等柱色谱及制备液相色谱进行分离,并根据有机波谱技术鉴定了化合物结构,进一步通过测定各个化合物对胰岛素抵抗的Hep G2细胞葡萄糖消耗量的影响以及黄酮苷类化合物对Akt磷酸化水平的影响来评价其抗糖尿病活性。结果从马齿苋中分离鉴定了9个化合物,分别为芹菜素-4'-O-α-L-鼠李糖苷(1),橙皮苷(2),山柰酚(3),木栓酮(4),6,7-二羟基香豆素(5),反式-对香豆酸(6),金莲花碱(7),咖啡酸(8),二十八烷酸(9),此外化合物1~4能够显著提高胰岛素抵抗Hep G2细胞葡萄糖消耗,其中化合物1的效果最为显著并能降低该细胞Akt的磷酸化水平。结论化合物1为首次从该植物中分离得到并且具有较强的抗糖尿病活性。     

桂枝茯苓胶囊化学成分研究(Ⅶ)

目的对桂枝茯苓胶囊内容物正丁醇萃取部位进行化学成分研究和活性筛选。方法采用多种色谱方法进行分离纯化,应用波谱数据解析鉴定化合物结构;对分离得到化合物进行原代小鼠子宫平滑肌细胞的钙离子内流作用的活性筛选。结果从桂枝茯苓胶囊内容物正丁醇层分离得到5个没食子酰苷类化合物,分别鉴定为3′-O-没食子酰蔗糖苷(1)、4′-O-没食子酰蔗糖苷(2)、6′-O-没食子酰蔗糖苷(3)、1′-O-没食子酰蔗糖苷(4)、1,2,3,4,6-五没食子酰葡萄糖(5)。结论化合物1～5为首次从该复方中分离得到,其中化合物1为新化合物;化合物1～5均有抑制原代小鼠子宫平滑肌细胞的钙离子内流作用。     

小桃儿七中2个新的甾体类化合物

目的 研究小桃儿七Helleborusthibetanus中的甾体类成分及其细胞毒性。方法 采用多种现代分离色谱技术进行分离纯化,根据MS、IR和NMR等数据鉴定新化合物的结构,采用MTT法对新化合物的细胞毒活性进行筛选。结果 从小桃儿七的正丁醇部位分离鉴定了2个甾体类化合物,分别鉴定为(25S)-27-O-β-D-葡萄吡喃糖基-3,25-二羟基螺甾-5(6)-烯-1β-yl-O-α-L-阿拉伯吡喃糖苷（1）和3β,14β-二羟基-3β-(2-羟基丙酰氧基)-5α-蟾酥甾-20,(22)-二烯（2）,化合物2对人结肠癌HCT116细胞、人肺癌A549细胞和人肝癌HepG2细胞的半数抑制浓度（half inhibitory concentration,IC50）分别为（10.8±0.4）、（4.8±0.7）、（45.3±1.8）μmol/L。结论 化合物1和2均为新化合物,分别命名为铁筷子苷M(thibetanosideM)和铁筷子苷N(thibetanoside N);化合物2表现出较好的细胞毒活性。     

石菖蒲乙醇提取物的化学成分及体外抗疲劳活性研究

采用柱色谱等分离手段,研究石菖蒲乙醇提取物中乙酸乙酯和正丁醇萃取部分的化学成分;并运用体外抗蟾蜍骨骼肌疲劳模型对分离得到的化合物进行抗疲劳活性研究.结果分离得到5个化合物,经波谱分析鉴定为细辛酮(1),环木菠萝烯酮(2),2,4,5-三甲氧基-2′-丁氧基-1,2-苯丙二醇(3),5-羟甲基糠醛(4),5-丁氧甲基糠醛(5).化合物3是新化合物,化合物2和5是首次从该植物中分离得到.化合物4具有明显的体外抗骨骼肌疲劳活性.     

双益方、组方中药及有效成分对骨骼肌及HepG2细胞胰岛素抵抗的影响

目的:研究双益方、组方中药及有效成分对骨骼肌细胞胰岛素抵抗模型葡萄糖消耗量及人肝癌HepG2细胞糖原合成,己糖激酶(hexokinase,HK),丙酮酸激酶(pyruvate kinase,PK)活性的影响,初步探讨其作用机制,为其临床应用奠定基础。方法:采用高胰岛素方法建立骨骼肌细胞的胰岛素抵抗(IR)模型,观察双益方低、中、高剂量组、组方中药(黄芪、山茱萸、黄连、桑白皮、葛根、佩兰),有效成分(黄芪甲苷,毛蕊异黄酮葡萄糖苷,熊果酸,马钱苷,小檗碱,1-脱氧野尻霉素,葛根素)组(30,120,480μg·L-1)对骨骼肌细胞胰岛素抵抗模型的葡萄糖消耗量的影响,蛋白质免疫印迹法(Western blot)测定骨骼肌细胞蛋白酪氨酸磷酸酶-1B(protein tyrosine phosphatase-1B,PTP-1B),葡萄糖转运蛋白4(glucosetransporters 4,GLUT4)表达;建立HepG2细胞的胰岛素抵抗模型,研究双益方、组方中药及其有效成分对HepG2细胞糖原合成及己糖激酶、丙酮酸激酶活性的影响。结果:双益方,黄芪,山茱萸,黄芪甲苷,1-脱氧野尻霉素,葛根素可降低骨骼肌细胞细胞上清液葡萄糖浓度(P0.05),以480μg·L-1双益方,黄芪甲苷组及葛根素组效果较好;与模型组比较,双益方、黄芪及有效成分组方可降低PTP-1B,提高GLUT-4的表达(P0.05,P0.01)。与正常组比较,模型组糖原合成量及HK,PK活性显著降低(P0.01);与模型组比较,各浓度组单味中药及有效成分均能一定程度的增加糖原合成量及HK,PK活性(P0.05,P0.01),其中黄芪甲苷、黄芪提取液增加糖原合成及提高HK活性作用较好;双益方提取液及葛根素增加PK活性作用较好。结论:双益方、组方中药及有效成分对骨骼肌,HepG2细胞胰岛素抵抗具有一定缓解作用,降低PTP-1B表达、提高GLUT4表达、增加糖原合成、提高HK,PK活性可能是其作用机制。     

玉米须化学成分研究

目的研究玉米须的化学成分及其细胞毒、葡萄糖消耗活性。方法采用溶剂萃取、硅胶柱色谱、高效液相色谱等方法进行分离纯化,通过理化性质及波谱数据分析鉴定结构,采用计算电子圆二色谱(ECD)方法确定新化合物的绝对构型;对分离得到的化合物1、2进行体外细胞毒及葡萄糖消耗活性筛选。结果从玉米须甲醇浸提液醋酸乙酯萃取物中分离得到4个化合物,分别鉴定为6β,12β-二羟基-4(5),7(8)-玉米二烯(1)、8α,10α-二羟基-4(5),7(12)-玉米二烯(2)、环己甲酸甲酯(3)、氮乙酰对羟基苯乙胺(4)。化合物1对人卵巢癌SK-OV-3细胞及膀胱癌T-24细胞的抑制率分别为11.86%、18.64%,对人肝癌HepG2细胞中葡萄糖消耗促进率为10.30%。结论化合物1、2是未见报道的新化合物,化合物1对SK-OV-3及T-24细胞显示抑制活性,对Hep G2细胞中葡萄糖消耗具有促进作用。     

α-Glucosidase inhibitory activity of selected Philippine plants

Ethnopharmacological relevance

Antidesma bunius Spreng. (Phyllantaceae), Averrhoa bilimbi L. (Oxalidaceae), Biophytum sensitivum (L.) DC. (Oxalidaceae), Ceriops tagal (Perr.) C.B. Rob. (Rhizophoraceae), Kyllinga monocephala Rottb. (Cyperaceae), and Rhizophora mucronata Lam. (Rhizophoraceae) are used as remedies to control diabetes. In the present study, these plants were screened for their potential α-glucosidase inhibitory activity.

Materials and methods

The 80% aqueous ethanolic extracts were screened for their α-glucosidase enzyme inhibitory activity using yeast alpha glucosidase enzyme.

Results

Except for A. bilimbi with IC₅₀ at 519.86±3.07, all manifested a significant enzyme inhibitory activity. R. mucronata manifested the highest activity with IC₅₀ at 0.08±1.82 μg mL⁻¹, followed by C. tagal with IC₅₀ at 0.85±1.46 μg mL⁻¹ and B. sensitivum with IC₅₀ at 2.24±1.58 μg mL⁻¹.

Conclusion

This is the first report on the α-glucosidase inhibitory effect of the six Philippine plants; thus, partly defining the mechanism on why these medicinal plants possess antidiabetic properties.     

European medicinal polypores – A modern view on traditional uses

Ethnopharmacological relevance

In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal–spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores.

Materials and methods

All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge.

Results

Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities.

Conclusions

The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.     

真菌的中药干预研究近5年进展

近年来真菌感染率逐年上升,传统抗真菌药物易产生耐药性,而中药在防治真菌感染方面具有一定的优势。本文就近5年来中药对白念珠菌、皮肤癣菌、曲霉菌、马拉色菌、串珠镰孢菌、申克孢子丝菌、新生隐球菌及真菌生物膜的干预研究进展进行综述。     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

列当属、肉苁蓉属和草苁蓉属植物传统药物学调查

通过野外资源调查、整理各大标本馆标本原始记录和查阅文献记载等方法,系统整理、总结、归纳了列当属、肉苁蓉属和草苁蓉属民族药用植物种类、功效及民间使用情况,结果表明列当属6种药用植物在4个少数民族间作为7种民族药应用,草苁蓉属2种药用植物在8个少数民族间作为10种民族药应用,肉苁蓉属2种药用植物在3个少数民族间作为3种民族药应用,且同种药用植物常作不同民族药;发现3属植物的传统疗效主要集中在补肾壮阳、止血和止痛3个方面,并且该传统疗效与现代药理研究结果基本吻合。因此深入研究植物种类丰富的列当属植物资源对缓解肉苁蓉植物资源匮乏局面和扩大药源具有积极意义。     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Comparative “in vitro” evaluation of the antiresorptive activity residing in four Ayurvedic medicinal plants. Hemidesmus indicus emerges for its potential in the treatment of bone loss diseases

Ethnopharmacological relevance

Four Indian plants, traditionally used in Ayurvedic medicine: Asparagus racemosus Willd., Emblica officinalis Gaertn., Hemidesmus indicus R. Br., and Rubia cordifolia L. were selected on the basis of their ethnobotanical use and of scientific evidence that suggests a potential efficacy in the treatment of bone-loss diseases. The antiresorptive properties of the four plants have been investigated. The aim was to provide adequate evidence for the exploitation of natural compounds as alternative therapeutics for the treatment of diseases caused by increased osteoclast activity.

Materials and methods

Decoctions were prepared from dried plant material according to the traditional procedure and standardization by HPLC was performed using marker compounds for each species. Total polyphenols, flavonoids and radical scavenging activity of the decoctions were also determined. The bioactivity of the plant decoctions was evaluated in subsequent phases. (1) A cytotoxicity screening was performed on the mouse monocytic RAW 264.7 cell line to define the concentrations that could be utilized in the following step. (2) The antiresorptive properties of plant decoctions were compared with that of a “gold standard” drug (alendronate) by measuring osteoclastogenesis inhibition and osteoclast apoptosis. (3) The toxic effect on bone forming cells was excluded by evaluating the impact on the proliferation of osteogenic precursors (mesenchymal stem cells, MSC).

Results

All the decoctions inhibited osteoclastogenesis similarly to alendronate at the highest doses, but Hemidesmus indicus and Rubia cordifolia were also effective at lower concentrations. Apoptosis increased significantly when cells were exposed to the highest concentration of Emblica officinalis, Hemidesmus indicus, and Rubia cordifolia. All concentrations of Emblica officinalis tested inhibited the proliferation of osteogenic precursors, while only the highest doses of Asparagus racemosus and Rubia cordifolia were toxic. On the contrary, Hemidesmus indicus did not affect osteogenic precursor growth at any concentration tested.

Conclusion

Among the medicinal plants included in the study, Hemidesmus indicus showed the greatest antiosteoclastic activity without toxic effect on osteogenic precursors. Therefore, Hemidesmus indicus exhibits the properties of an antiresorptive drug and represents the ideal candidate for further clinical investigations.     

金银花类药用植物IspE和IspH基因克隆和生物信息学分析

目的:克隆金银花类药用植物4-二磷酸胞苷-2-C-甲基赤藓糖激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,IspE)和4-羟基-3-甲基-2-邻苯基二磷酸还原酶(4-hydroxy-3-methylbut-2-enyl diphosphate reductase,IspH)基因,并对其基因序列、蛋白特性和转录活性进行分析、比较.方法:从忍冬Lonicera japonica转录组测序结果中分析获得了IspE,IspH基因.分别以忍冬、红白忍冬L.japonica var.chinensis、红腺忍冬L.hypoglauca和水忍冬L.dasystyla新鲜花蕾为材料,利用RT-PCR技术克隆获得了4种金银花类药用植物IspE和IspH基因的全长cDNA.运用生物信息学分析软件,预测编码蛋白的结构和功能,并通过RT-PCR检测IspE和IspH在忍冬、红白忍冬、红腺忍冬、水忍冬花蕾中的转录情况.结果:金银花类药用植物IspE基因含有1个完整的开放阅读框,长度为1 221 bp,编码406个氨基酸;IspH含有一个完整的开放阅读框,长度为1 380 bp,编码459个氨基酸.IspE和IspH均为非分泌蛋白,均定位于叶绿体中.RT-PCR分析结果表明在忍冬、红腺忍冬和水忍冬的花蕾中IspE,IspH基因的转录水平没有显著差异,但红白忍冬花蕾中IspE,IspH基因的转录水平均显著高于忍冬.结论:克隆获得忍冬、红白忍冬、红腺忍冬和水忍冬中IspE,IspH基因,并证实了其在不同金银花类药用植物中的表达,为进一步研究IspE,IspH基因对萜类化合物生物合成和花香气以及颜色的影响奠定了基础.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.