In vitro effects of Sutherlandia frutescens water extracts on cell numbers,morphology, cell cycle progression and cell death in a tumorigenic and a non-tumorigenic epithelial breast cell line

Sutherlandia frutescens is a South African herb traditionally used for internal cancers, diabetes, a variety of inflammatory conditions and recently to improve the overall health in cancer and HIV/AIDS patients. The in vitro effects of S. frutescens extracts were evaluated on cell numbers, morphology, cell cycle progression and cell death. Dose-dependent studies (2–10 mg/ml) revealed a decrease in malignant cell numbers when compared to their controls. S. frutescens extracts (10 mg/ml) decreased cell growth in a statistically significantly manner to 26% and 49% (P < 0.001) in human breast adenocarcinoma (MCF-7) and human non-tumorigenic epithelial mammary gland cells (MCF-12A) respectively after 72 h of exposure. Cell density was significantly compromised and hypercondensed chromatin, cytoplasmic shrinking, membrane blebbing and apoptotic bodies were more pronounced in the MCF-7 cell line. Both S. frutescens-treated cell lines exhibited and increased tendency for acridine orange staining, suggesting increased lysosomal and/or autophagy activity. Flow cytometry showed an increase in the sub G₁ apoptotic fraction and an S phase arrest in both the 5 mg/ml and 10 mg/ml S. frutescens-treated cells. S. frutescens induced an increase in apoptosis in both cell lines as detected by Annexin V and propidium iodide flow cytometric measurement. At 10 mg/ml, late stages of apoptosis were more prominent in MCF-7 S. frutescens-treated cells when compared to the MCF-12A cells. Transmission electron microscopy revealed hallmarks of increased vacuolarization and hypercondensed chromatin, suggesting autophagic and apoptotic processes. The preliminary study demonstrates that S. frutescens water extracts exert a differential action mechanism in non-tumorigenic MCF-12A cells when compared to tumorigenic MCF-7 cells, warranting future studies on this multi-purpose medicinal plant in southern Africa.     

Influence of Sutherlandia frutescens extracts on cell numbers, morphology and gene expression in MCF-7 cells

Sutherlandia frutescens is a well-known South African herbal remedy traditionally used for stomach problems, internal cancers, diabetes, various inflammatory conditions and recently to improve the overall health in cancer and HIV/AIDS patients. The influence of crude Sutherlandia frutescens extracts (prepared with 70% ethanol) was investigated on cell numbers, morphology, and gene expression profiles in a MCF-7 human breast adenocarcinoma cell line. Time-dependent (24, 34, 48 and 72 h) and dose-dependent (0.5-2.5 mg/ml) studies were conducted utilizing spectrophotometrical analysis with crystal violet as DNA stain. A statistically significant decrease to 50% of malignant cell numbers was observed after 24 h of exposure to 1.5 mg/ml Sutherlandia frutescens extract when compared to vehicle-treated controls. Morphological characteristics of apoptosis including cytoplasmic shrinking, membrane blebbing and apoptotic bodies were observed after 24h of exposure. A preliminary global gene expression profile was obtained by means of microarray analysis and revealed valuable information about the molecular mechanisms and signal transduction associated with 70% ethanolic Sutherlandia frutescens extracts.     

Cyclocarya paliurus extract modulates adipokine expression and improves insulin sensitivity by inhibition of inflammation in mice

Ethnopharmacological relevance

Cyclocarya paliurus Batal., a Chinese native plant, is the sole species in its genus and its leaves have been widely used as a remedy for diabetes in traditional folk medicine. The study was undertaken to evaluate the effects of Cyclocarya paliurus leaves extracts (CPE) on adipokine expression and insulin sensitivity in mice.

Materials and methods

Mice were stimulated with conditioned medium (prepared from activated macrophages, Mac-CM) to induce adipose dysfunction and insulin resistance. Then mice were treated with CPE (100, 200 and 500 mg/kg, ig.) or metformin (200 mg/kg, ig.), followed by glucose and insulin intolerance, adipokine expression, phosphorylation of insulin receptor substrate (IRS-1) and glucose consumption measurement.

Results

CPE, as well as metformin effectively promoted glucose disposal in oral glucose tolerance test in normal mice. Mac-CM challenge induced glucose and insulin intolerance, but CPE reversed these alternations with increased glycogen content in muscle and liver, well demonstrating its beneficial effects on glucose homeostasis. RT-qPCR analysis showed that CPE inhibited TNF-a, IL-6, MCP-1 and resistin overexpression and effectively enhanced adiponectin expression in adipose tissue when mice were exposed to Mac-CM stimulation. Inflammation impaired insulin signaling in muscle, whereas CPE inhibited inflammation-induced serine phosphorylation of IRS-1 and effectively restored the phosphorylation of both IRS-1 at tyrosine residues and downstream Akt phosphorylation in response to insulin. Moreover, independently of insulin, CPE promoted glucose consumption in adipocytes under normal and inflammatory conditions.

Conclusion

Above-mentioned results demonstrated that CPE beneficially regulated adipokines expression and ameliorated insulin resistance through inhibition of inflammation in mice.     

白木香乙酰乙酰基辅酶A硫解酶基因(AsAACT)的克隆与表达分析

目的：对国产沉香的基原植物白木香Aquilaria sinensis （Lour.）Gilg乙酰辅酶A酰基转移酶（acetyl-CoA C-acetyl transferase,AACT）基因AsAACT全长进行克隆并展开生物信息学分析和表达分析,为解析沉香萜类次生代谢产物的生物合成机制奠定基础。方法：根据获得的白木香转录组数据库AACT部分转录本序列设计引物,采用RT-PCR及RACE技术,以白木香茎cDNA为模板,克隆获得AsAACT全长,进行生物信息学分析;采用荧光定量PCR,以GADPH为内参,分析白木香愈伤组织受不同伤害胁迫AsAACT的表达模式。结果：AsAACT开放阅读框（opening reading frame,ORF）为1 236 bp,编码411个氨基酸残基,酶命名为AsAACT;白木香愈伤受物理伤害（切割）后表达量没有明显变化,但受化学伤害（MeJA）后4 h表达量升高了5.5倍,说明该基因对MeJA诱导的化学伤害较敏感,且能够在早期响应伤害胁迫。结论：通过AsAACT基因的全长cDNA克隆和表达特性分析,为后续深入研究其在沉香倍半萜合成途径的功能奠定基础。     

Differential signaling involved in Sutherlandia frutescens-induced cell death in MCF-7 and MCF-12A cells

Ethnopharmacological relevance

The scientific study of natural products traditionally used in anticancer preparations has yielded several therapeutically relevant compounds. One of these traditional preparations with potentially beneficial properties is aqueous extracts of Sutherlandia frutescens, a shrub indigenous to the Western Cape region of South Africa. The aims of this study were to evaluate in vitro efficacy of these preparations on the MCF-7 breast adenocarcinoma and MCF-12A non-tumorigenic cell lines in terms of cell proliferation, cell morphology and possible induction of cell death.

Materials and methods

Crystal violet staining was used to evaluate cell proliferation, light-and fluorescence microscopy were used to investigate both intracellular and extracellular morphological features of apoptosis and autophagy (e.g. membrane blebbing, condensed chromatin and intracellular lysosomes), while flow cytometry quantified cell cycle changes and induction of apoptosis through analysis of the flip-flop translocation of phosphatidylserine.

Results

Crystal violet staining showed a time- and dose specific response to aqueous Sutherlandia frutescens extracts, revealing exposure to 1 mg/ml aqueous extract for 48 h to be ideal for comparing the differential effects of Sutherlandia frutescens in the MCF-7 and MCF-12A cell lines. Microscopy showed distinct morphological changes with hallmarks of apoptosis being observed in both cell lines. Flow cytometry revealed a decrease in actively cycling cells in both cell lines, and a 4.36% increase in phosphatidylserine translocation in the MCF-7 cell line, indicative of apoptosis induction, while fluorescence microscopy showed evidence of the induction of autophagy.

Conclusions

Analyses revealed the carcinogenic MCF-7 cell line to be more susceptible to the cytostatic and cytotoxic effects of aqueous extracts of Sutherlandia frutescens when compared to the non-tumorigenic MCF-12A cell line, thus warranting further research into the exact cellular mechanisms involved and the possible synergistic activities of Sutherlandia frutescens ingredients.     

ZnSO4和PEG引发对老化紫苏种子萌发及 幼苗抗氧化酶活性的影响

目的: 通过对老化的紫苏种子萌发及幼苗生理特性的研究,寻找提高紫苏种子及幼苗抗老化能力的途径。 方法 :测定不同浓度硫酸锌(ZnSO4)和聚乙二醇(PEG)处理后的紫苏种子的发芽势、发芽率、发芽指数和活力指数,并对紫苏幼苗叶片的丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性进行测定。 结果 :老化的紫苏种子在经过不同浓度的ZnSO4和PEG处理后,萌发指标均有升高,其中经过600 mg·L^-1的ZnSO4和20%的PEG处理后,各项指标均达到最大值,发芽势(Gv)分别为64.7,66.8%;发芽率 (Gr) 分别为78.7%,79.4%;发芽指数(Gi) 分别为11.8,12.2;活力指数(Vi) 分别为0.091 1,0.093 9。不同处理均降低了叶片MDA的含量,并且提高了SOD,POD,CAT 3种酶的活性,显著的提高了幼苗生物量的积累。当ZnSO4为600 mg·L^-1时, 3种酶活性分别为0.28,4.71, 3.82 U·mg^-1,当PEG浓度为20%时,3种酶的活性分别为0.29,4.93,4.18 U·mg^-1。 结论 :600 mg·L^-1的ZnSO4和20%的PEG处理能够有效的减缓老化对紫苏种子及幼苗产生的伤害,提高了种子及幼苗的抗老化能力。     

地黄Aux/IAA家族基因RgIAA1的克隆和表达分析

为了克隆地黄Aux/IAA家族基因RgIAA1,分析地黄RgIAA1的时空表达特性及对逆境胁迫的响应。以拟南芥AtIAA14的cDNA序列为探针在地黄转录组数据库中进行比对,应用生物信息学方法分析RgIAA1的序列特征及进化关系,以实时荧光定量PCR技术检测RgIAA1在地黄不同组织以及逆境胁迫下的表达量。结果表明,获得1条903 bp的cDNA序列,包含一个681 bp完整的开放阅读框（ORF）,编码226个氨基酸,具有Aux/IAA家族蛋白的典型结构域和特征序列。RgIAA1在地黄不同组织中的表达呈差异表达,其中在地黄幼嫩叶片里表达强度最大,其次为茎,且随着叶片的伸展,RgIAA1表达强度不断降低。在连作时RgIAA1的表达量升高,NaCl和渍水胁迫下RgIAA1的表达量降低。实验首次克隆了地黄Aux/IAA家族基因RgIAA1,为阐明其在地黄生长发育和逆境胁迫中的分子功能奠定基础。     

北柴胡糖基转移酶基因BcUGT3,BcUGT6的表达分析及其原核表达

该研究测定了北柴胡中2个糖基转移酶基因BcUGT3和BcUGT6在不同组织中的转录水平及MeJA处理后不定根中的转录水平。BcUGT3在根、叶、花和果中的转录水平无明显差异,但均高于茎中的转录水平。BcUGT6在叶中转录水平最高,在花中最低。以未处理的为对照,BcUGT6在MeJA处理后2 h,8 h,24 h,2 d,4 d的北柴胡不定根中,转录水平均提高近2倍,表明BcUGT6的转录受MeJA影响较小。BcUGT3转录水平随处理时间的延长不断增加,4 d时,达7倍左右。利用载体pET-28a（+）,进行了2个基因的原核表达。IPTG诱导后,宿主菌表达出了目的蛋白,并获得了纯化蛋白。为后续开展这2个基因的功能研究奠定了基础。     

滇重楼鲨烯合酶基因PpSQS的克隆及在大肠杆菌中的表达

目的: 克隆滇重楼鲨烯合酶(squalene synthase,SQS)基因(PpSQS),对该基因进行序列分析及原核表达。 方法: 采用同源克隆和RACE技术获得PpSQS的cDNA,并对其进行生物信息学分析;构建原核表达载体pET-30b(+)-PpSQS,在大肠杆菌Escherich coli BL21(DE3)中进行诱导表达。 结果: PpSQS基因cDNA全长1 498 bp,包含1个1 212 bp的开放阅读框(ORF),可编码403个氨基酸;PpSQS理论标准分子质量为46.36 kDa,等电点为pI 6.83;SDS-PAGE分析表明,经1 mmol·L^-1 IPTG诱导后,重组PpSQS蛋白在大肠杆菌中获得表达。 结论: 首次获得了滇重楼PpSQS基因cDNA全长序列,该基因编码产物具有植物SQS同源蛋白的典型特征,实现重组PpSQS在大肠杆菌中的表达。     

金荞麦苯丙氨酸解氨酶基因FdPAL 的克隆及原核表达

目的:克隆金荞麦苯丙氨酸解氨酶(FdPAL)基因DNA和全长cDNA序列,对该基因进行序列分析,原核表达及其活性相关研究.方法:采用同源克隆和RT-PCR技术扩增苯丙氨酸解氨酶基因的DNA序列和全长cDNA序列并对其进行生物信息学分析;构建PAL原核表达载体pET30b(+)-FdPAL,在大肠杆菌Escherich coli BL21( DE3)中进行诱导表达,使用分光光度计法定量测定其酶活,使用薄层色谱法鉴定其催化活性.结果:金养麦PAL基因DNA序列全长2 583 bp,由2个外显子,1个内含子构成(命名为FdPAL,GenBank登录号为HM628904);cDNA序列包含1个2 169 bp的开放阅读框(ORF),编码722个氨基酸,理论相对标准分子质量78.31 kDa,等电点5.94.SDS-PAGE分析表明,诱导后的新生蛋白质相对标准分子质量为75.37 kDa;诱导4h后,表达产物的苯丙氨酸解氨酶比活力最高,达到4 386 nmol·g-1·min-;薄层色谱结果表明,诱导表达产物具有催化苯丙氨酸转化为肉桂酸的活性.结论:首次从金荞麦中克隆到PAL基因,该基因具有植物PAL同源基因的典型特征;重组的金荞麦PAL基因表达载体[pET30b(+)-FdPAL]能有效地在大肠杆菌Ecoli BL21 (DE3)中表达,并形成具有一定催化功能的酶.     

Scoparia dulcis (SDF7) endowed with glucose uptake properties on L6 myotubes compared insulin

Aim of the study

Insulin stimulates glucose uptake and promotes the translocation of glucose transporter 4 (Glut 4) to the plasma membrane on L6 myotubes. The aim of this study is to investigate affect of Scoparia dulcis Linn water extracts on glucose uptake activity and the Glut 4 translocation components (i.e., IRS-1, PI 3-kinase, PKB/Akt2, PKC and TC 10) in L6 myotubes compared to insulin.

Materials and methods

Extract from TLC fraction-7 (SDF7) was used in this study. The L6 myotubes were treated by various concentrations of SDF7 (1 to 50 μg/ml) and insulin (1 to 100 nM). The glucose uptake activities of L6 myotubes were evaluated using 2-Deoxy-D-glucose uptake assay in with or without fatty acid-induced medium. The Glut 4 translocation components in SDF7-treated L6 myotubes were detected using immunoblotting and quantified by densitometry compared to insulin. Plasma membrane lawn assay and glycogen colorimetry assay were carried out in SDF7- and insulin-treated L6 myotubes in this study.

Results

Here, our data clearly shows that SDF7 possesses glucose uptake properties on L6 myotubes that are dose-dependent, time-dependent and plasma membrane Glut 4 expression-dependent. SDF7 successfully stimulates glucose uptake activity as potent as insulin at a maximum concentration of 50 μg/ml at 480 min on L6 myotubes. Furthermore, SDF7 stimulates increased Glut 4 expression and translocation to plasma membranes at equivalent times. Even in the insulin resistance stage (free fatty acids-induced), SDF7-treated L6 myotubes were found to be more capable at glucose transport than insulin treatment.

Conclusions

Thus, we suggested that Scoparia dulcis has the potential to be categorized as a hypoglycemic medicinal plant based on its good glucose transport properties.     

Essential oil of Perilla frutescens-induced change in hippocampal expression of brain-derived neurotrophic factor in chronic unpredictable mild stress in mice

Ethnopharmacological relevance

Perilla frutescens (Perilla leaf), a traditional Chinese medicinal herb, has been used for centuries to treat various conditions including depression. A previous study of the authors demonstrated that essential oil of Perilla frutescens (EOPF) attenuated the depressive-like behavior in mice.

Aim of the study

This study was undertaken to explore the dynamic change of behaviors and brain-derived neurotrophic factor (BDNF) expression induced by chronic unpredictable mild stress (CUMS), and improved by EOPF.

Materials and methods

Four separate CUMS experimental groups (1-week, 2-week, 3-week and 4-week treatment) were treated with EOPF (3 mg/kg and 6 mg/kg, p.o.) or fluoxetine (20 mg/kg, p.o.), followed by sucrose preference, locomotor activity, immobility and hippocampal BDNF measurement.

Results

EOPF, as well as fluoxetine, restored the CUMS-induced decreased sucrose preference and increased immobility time, without affecting body weight gain and locomotor activity. Furthermore, CUMS (3 or 4-week) produced a reduction in both BDNF mRNA and protein expression in the hippocampus, which were ameliorated by EOPF (4-week) and fluoxetine (3 or 4-week) treatment.

Conclusion

These results presented here show that BDNF is expressed depending on length of CUMS procedure and EOPF administration. And this study might contribute to the underlying reason for the slow onset of antidepressant activity in clinic.     

Beneficial metabolic effects of the Malaysian herb Labisia pumila var. alata in a rat model of polycystic ovary syndrome

Aim of the study

New options are needed to prevent and treat metabolic disorders associated with polycystic ovary syndrome (PCOS). Labisia pumila var. alata (LPva)—a Malaysian herb thought to have phytoestrogenic effects—has shown promise in reducing body weight gain in ovariectomized rats. In this study, we investigated the effect of LPva on body composition and metabolic features in female rats treated continuously with dihydrotestosterone, starting before puberty, to induce PCOS.

Material and methods

At 9 weeks of age, the PCOS rats were randomly subdivided into two groups; PCOS LPva and PCOS control. PCOS LPva rats received a daily oral dose of LPva (50 mg/kg body weight), dissolved in 1 ml of deionised water, for 4–5 weeks. PCOS controls received 1 ml of deionised water on the same schedule.

Results

LPva increased uterine weight (27%) and insulin sensitivity (36%) measured by euglycemic hyperinsulinemic clamp. Plasma resistin levels were increased and lipid profile was improved in LPva rats. In adipose tissue, LPva decreased leptin mRNA expression but did not affect expression of resistin and adiponectin. No effects on body composition, adipocyte size, or plasma leptin levels were observed.

Conclusion

LPva increases uterine weight, indicating estrogenic effects, and improves insulin sensitivity and lipid profile in PCOS rats without affecting body composition.     

丹参转录因子SmbHLH1基因的克隆和表达分析

目的:获得丹参转录因子SmbHLH1全长基因,分析SmbHLH1基因在丹参不同组织部位,以及不同诱导子处理后的表达差异.方法:利用RT-PCR等技术获得SmbHLH1基因全长,利用半定量RT-PCR,分析SmbHLH1基因在丹参不同部位的表达情况,及在不同处理条件下的表达情况.结果:获得的SmbHLH1基因由999个核苷酸组成,编码332个氨基酸,蛋白相对分子质量约36 kDa;半定量RT-PCR检测,该基因在丹参的根中表达量最高,其次是茎,在叶和花中有微量表达;茉莉酸甲酯,以及酵母提取物和Ag+共同诱导,会抑制该基因的表达,水杨酸和脱落酸对该基因的影响不大.结论:丹参SmbHLH1基因是bHLH家族新成员,其功能可能与油菜素内酯信号途径有关.     

刺五加钙调蛋白基因的克隆及内生真菌对其表达的影响

目的:克隆刺五加的钙调蛋白(calmodulin,CaM)基因,并分析内生真菌对其表达的影响。方法:采用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术克隆刺五加CaM基因的全长cDNA序列。通过RT-PCR法检测可显著提高刺五加皂苷含量的内生真菌菌株P116-1a,P116-1b,P109-4,P312-1对CaM表达的影响。结果:刺五加CaM基因的cDNA全长为856 bp,开放阅读框长450 bp,编码149个氨基酸的蛋白,与人参Panax ginseng和胡萝卜Daucus carota等物种的CaM同源性均高达100%。RT-PCR的结果显示,内生真菌可显著提高刺五加CaM基因的表达量(P<0.05),最大表达量出现在菌株P109-4回接90 d时,达对照的2.96倍。结论:首次克隆并报道了刺五加CaM的cDNA全长序列,并证实内生真菌可显著提高刺五加CaM基因的表达量,为阐明内生真菌提高刺五加三萜皂苷含量的机制奠定了基础。     

铁皮石斛HSP70基因的克隆及冷胁迫表达分析

目的: 揭示铁皮石斛热激蛋白HSP70基因冷胁迫表达,为耐低温品系选育提供分子生物学基础。 方法: 根据已获得的HSP70基因片段序列,采用RACE方法从铁皮石斛中克隆到HSP70基因全长cDNA序列。预测其编码蛋白的结构与功能,并运用实时定量PCR进行冷胁迫下表达分析。 结果: 核苷酸序列分析表明该基因全长2 296 bp,包含一完整的1 944 bp的开放阅读框,编码647个氨基酸。氨基酸序列具有典型的HSP70特征并且与其他植物的HSP70有很高的同源性。冷胁迫表达分析说明该基因确实能被低温诱导表达。 结论: 首次克隆获得受低温诱导表达的铁皮石斛HSP70基因,为进一步研究铁皮石斛健康栽培与抗寒品系的选育奠定基础。     

白木香法呢基焦磷合酶基因AsFPS1的克隆及表达分析

法呢基焦磷酸合酶(FPS) 是倍半萜代谢途径中的关键限速酶之一.本研究在454高通量测序已获得的cDNA序列基础上,PCR扩增其开放阅读框并测序验证;提取三年生白木香根、茎、叶的总RNA及健康和伤害处理的愈伤组织的总RNA,以反转录成的单链cDNA为模板进行荧光定量PCR,检测AsFPS1基因的组织表达特异性及对伤害处理的反应.结果表明,扩增到的AsFPS1基因全长1 342 bp,开放阅读框1 029 bp,编码342个氨基酸.组织表达分析的结果显示,AsFPS1基因主要在根和茎中表达,叶中的表达量较低;该基因同时受物理伤害和结香液处理的诱导,分别在6,12 h达到最高表达水平.通过AsFPS1基因全长cDNA的克隆和表达分析,为后续研究其生物功能及沉香倍半萜合成机制奠定了基础.     

铁皮石斛蔗糖合成酶基因的克隆及表达分析

目的:克隆铁皮石斛Dendrobium officinale蔗糖合成酶基因并对其表达分析,为研究铁皮石斛多糖合成与蔗糖合成酶活性关系及调控表达提供理论依据.方法:基于GenBank上已知的蔗搪合成酶基因SS同源序列,应用RT-PCR和RACE等方法,克隆铁皮石斛蔗糖合成酶基因全长,将目的片段转化大肠杆菌表达菌株BL21中,IPTG诱导表达,SDS-PAGE电泳分析.结果:成功克隆了铁皮石斛蔗糖合成酶基因,该基因cDNA全长2 424 bp,编码807个氨基酸(命名为DOSS1),登录号HQ856835,该基因氨基酸序列与兰科植物Moluara yellow(AF530568)同源性达到95%,与Oncidium goldiana(AF530567)同源性达到90%,与禾本科植物的SS基因同源性在80%以上.并对其进行大肠杆菌原核表达诱导,诱导的工程菌能明显表达出一条相对分子质量约为92×10(3)左右的蛋白质,与理论相对分子质量基本相符.结论:成功克隆了铁皮石斛蔗糖合成酶基因,并诱导出一条与理论蛋白相对分子质量相符的蛋白条带.     

Effects of Gelsemium sempervirens L. on pathway-focused gene expression profiling in neuronal cells

Ethnopharmacological relevance

Gelsemium sempervirens L. is a traditional medicinal plant mainly distributed in the southeastern of the United States, employed in phytotheraphy and homeopathy as nervous system relaxant to treat various types of anxiety, pain, headache and other ailments. Although animal models showed its effectiveness, the mechanisms by which it might operate on the nervous system are largely unknown. This study investigated for the first time by a real-time PCR technique (RT-PCR Array) the gene expression of a panel of human neurotransmitter receptors and regulators, involved in neuronal excitatory signaling, on a neurocyte cell line.

Materials and methods

Human SH-SY5Y neuroblastoma cells were exposed for 24 h to Gelsemium sempervirens at 2c and 9c dilutions (i.e. 2 and 9-fold centesimal dilutions from mother tincture) and the gene expression profile compared to that of cells treated with control vehicle solutions.

Results

Exposure to the Gelsemium sempervirens 2c dilution, containing a nanomolar concentration of active principle gelsemine, induced a down-regulation of most genes of this array. In particular, the treated cells showed a statistically significant decrease of the prokineticin receptor 2, whose ligand is a neuropeptide involved in nociception, anxiety and depression-like behavior.

Conclusions

Overall, the results indicate a negative modulation trend in neuronal excitatory signaling, which can suggest new working hypotheses on the anxiolytic and analgesic action of this plant.     

Extract isolated from Angelica hirsutiflora with insulin secretagogue activity

Ethnopharmacological relevance

Angelica genus (Umbelliferae) has traditionally been used as the medicine and health food considered alleviating several disorders including diabetes mellitus. Angelica hirsutiflora Liu Chao & Chuang is an endemic species and a folk medicine in Taiwan.

Aim of the study

The scientific evidence of anti-diabetic effect for Angelica hirsutiflora remains unknown. The methanolic extracts isolated from Angelica hirsutiflora were studied for its insulin secretagogue and hypoglycemic activities.

Materials and methods

The in vitro effects and possible mechanisms of Angelica hirsutiflora extract on the insulin secretion in isolated mouse and human islets and pancreatic β-cell line HIT-T15 were determined; and tested the regulation of blood glucose in fasted mice and high-fat diet-induced diabetic mice.

Results

Angelica hirsutiflora extract potently stimulated the release of insulin from cultured HIT-T15 cells and isolated mouse and human islets. The intracellular calcium levels were also increased in HIT-T15 cells and isolated human islets treated with Angelica hirsutiflora extract. Angelica hirsutiflora extract was capable of enhancing the phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2 protein in HIT-T15 cells. Specific ERK inhibitor PD98059 inhibited the increase of insulin secretion by Angelica hirsutiflora extract in HIT-T15 cells and isolated mouse islets. When Angelica hirsutiflora extract was administered to the fasted mice, it decreased the rise in blood glucose level after starch loading. The plasma insulin level was also increased by Angelica hirsutiflora extract treatment. In high-fat diet-induced diabetic mice, Angelica hirsutiflora extract markedly improved the oral glucose intolerance as compared with the vehicle control.

Conclusions

These findings support that Angelica hirsutiflora extract may be useful in the control of hyperglycemia in non-insulin-dependent diabetes mellitus by acting as an insulin secretagogue.