Two pyrrole acids isolated from Phyllanthus emblica L.and their bioactivities

An undescribed pyrrole acid,1-(4′-methoxy-4′-oxobutyl)-1 H-pyrrole-2,5-dicarboxylic acid(1)and one known pyr-role acid(2)were isolated from the fruits of Phyllanthus emblica.The structures of these compounds were elucidated via the comprehensive analyses of IR,HRESIMS,1D and 2D spectroscopic data.A series of biological assays revealed that compounds 1 and 2 could inhibit LPS-induced over-production of nitric oxide(NO),interleukin-6(IL-6),monocyte chemotactic protein 1(MCP-1)and tumor necrosis factor-α(TNF-α)by reducing the phosphorylation of extracellular regulated protein kinases(ERK)and c-Jun N-terminal kinases(JNK)in RAW 264.7 cells.Additionally,compounds 1 and 2 were found to reduce lipid deposition and increase the mRNA expression of ATP-binding cassette transporter A1 in oxidized low-density lipoprotein-treated RAW264.7 macrophages.

Phytochemistry and ethnomedicinal qualities of metabolites from Phyllanthus emblica L.:A review

Phyllanthus emblica or Indian gooseberry is an integrated part of Ayurvedic and Traditional Chinese Medicines.For several decades,the well-known ancient herb has been extensively utilized in traditional medicine to cure diseases like fever,diabetes,constipation,jaundice,ulcers,biliousness,anemia,anorexia,and dyspepsia.In the traditional system,Indian gooseberry has various ethnomedicinal applications.In the Ayurvedic system,different methods of administration(anupan)have shown different ethnomedicinal properties of Indian gooseberry.Seventy well-known chemical components in Indian gooseberry have been identified through phytochemical evaluation,among which the flavonoids and phenols are most prominent.From the toxicity perspective,it is considered a safe herb in India,and is taken as a food supplement in European countries.The wide-spectrum pharmacological activities of the crude extracts and isolates of Indian gooseberry are attributed to the predominance of phenols and flavonoids.Thus,it is important to study the exact mechanism of the activity of the phytochemicals in Indian gooseberry,especially in anti-cancer activities.Extract of Indian gooseberry enhances proliferation in several cancer cells in vitro,including stem cells like ovarian cancer(OC)cells,and also has been observed to possess anti-proliferative characteristics in vivo.This review intends to explore the therapeutic potential of Indian gooseberry based on scientific reports and attempts to find the gaps for future research.

余甘子(Phyllanthus emblica L.)Vc含量分析

采用2,6-二氯酚靛酚滴定法分析比较了余甘子各器官之间、同品种不同植株之间、不同余甘子品种之间以及与野生资源之间的Vc含量以及不同预处理方法对Vc测定的影响。结果表明,Vc在余甘子各器官中均有分布,以果实与初生幼叶中居多,茎、根中含量较低;皇帝甘与粉甘含量较多,但与其它品种的差异不大;5个野生资源单株Vc含量差异极显著,其中有3个野生单株的果肉Vc含量高于主栽品种粉甘。

余甘(Phyllanthus emblica L.)大小孢子发生及雌雄配子体发育研究

采用常规石蜡切片技术,对平丹1号余甘大小孢子发生及雌雄配子体发育进行研究。结果表明:药壁发育为双子叶型;绒毡层为腺质绒毡层;小孢子母细胞减数分裂时胞质分裂为同时型;产生四面体形四分体;成熟的花粉粒为二细胞型花粉;平丹1号余甘的雌蕊为上位子房,3心皮3室,每室含倒生胚珠2枚,为厚珠心,具珠心喙,胚囊为蓼形;平丹1号余甘部分大孢子母细胞、大孢子及胚囊发育异常,致使不能形成正常的卵细胞,不能进行受精。

Anti-diabetic activity of quercetin extracted from Phyllanthus emblica L.fruit: In silico and in vivo approaches

In this study, molecular interactions of the ligands, quercetin, gallic acid, and metformin with various diabetes mellitus-related protein targets, such as glycogen phosphorylase and peroxisome proliferatoractivated receptor gamma, were assessed. It was revealed that quercetin possesses good binding affinity to both targets. Quercetin is a major constituent of methanolic extracts of Phyllanthus emblica fruit. The antihyperglycemic effect of quercetin in streptozotocin(STZ)-induced diabetic rats was examined. The isolated quercetin administered at a dose of 75 mg/kg body weight produced a maximum decrease of14.78% in blood glucose levels in the diabetic rats after 7 days of treatment. Furthermore, quercetin doses of 50 and 75 mg/kg were shown to significantly improve the profiles of triglycerides, high-density lipoprotein, very-low-density lipoprotein, low-density lipoprotein, and total cholesterol at the end of the study in STZ-induced diabetic rats. The administration of quercetin(25, 50, and 75 mg/kg body weight)daily for 28 days in STZ-induced diabetic rats resulted in a significant decrease in blood glucose and urine sugar levels, with a considerable rise in plasma insulin and hemoglobin levels. Therefore, quercetin is a potential drug with antidiabetic and antihyperglycemic action mediated by changes in the levels of glucose, cholesterol, and triglycerides as indicated by in silico and in vivo studies.

Effect of Total Flavonoids from Phyllanthus emblica L. on Tumor Proliferation and Immune Function

[Objectives]To explore the effect of total flavonoids from Phyllanthus emblica L.on tumor proliferation and immune function.[Methods]The effects of total flavonoids of P.emblica L.on the proliferation of 6 different tumor cell lines(human hepatoma cell line HepG-2,human cervical cancer cell line Hela,human gastric cancer cell line SGC7901,human nasopharyngeal carcinoma cell line CNE-2,human lung cancer cell line H460 and human ovarian cancer cell line A2780)were compared by SRB method.The effect of total flavonoids from P.emblica L.on the proliferation of mouse lymphocytes induced by concanavalin(ConA)or lipopolysaccharide(LPS)in vitro was detected by CCK-8 method.[Results]The results of SRB assay showed that compared with the normal group,the total flavonoids of P.emblica L.had obvious inhibitory effect on 6 kinds of tumor cells.Among them,the inhibitory effect on H460 cells and CNE-2 cells was the most significant,and the IC50 was(471.36±50.66),(463.26±40.75)μg/mL,respectively.The high dose group of total flavonoids from P.emblica L.had the same inhibitory effect as the positive drug 5-Fu.The results of CCK-8 assay showed that compared with the blank group,the total flavonoids of P.emblica L.significantly inhibited the proliferation of mouse lymphocytes induced by ConA or LPS(*P<0.05,**P<0.01).[Conclusions]The total flavonoids of P.emblica L.had significant anti-tumor activity.

Phyllanthuse emblica polyphenols:Optimization of high-voltage pulsed electric field assisted extraction,an antioxidant and anti-inflammatory effects in vitro

Background:The polyphenols extraction of Phyllanthus emblica is primarily carried out using organic solvents,and assisted by physical fields such as ultrasound and microwave for extraction.High voltage pulsed electric field technology(PEF)is a non-thermal processing technology that has high efficiency and minimal damage to thermosensitive substances.PEF has been applied to plant extraction in many studies,however,the extraction of polyphenols from Phyllanthus emblica using the PEF has still not been reported;Objective:This study explores the optimal extraction process of polyphenols from Phyllanthus emblica using the PEF,and investigates its relaxation and anti-wrinkle based on anti-oxidation and anti-inflammatory experiment,in order to develop a Phyllanthus emblica extract with substantial efficacy;Materials and Methods:The method of Phyllanthus emblica extract using PEF is established,and compared with a traditional extraction method.The experimental conditions,such as electric field intensity(0.5–6.0 kV/cm),pulse times(20−120),extraction time(0–60 min)and material concentration(0.5%∼3%),are investigated and optimized using orthogonal experiments;Results:the polyphenols in the Phyllanthus emblica extract were highest at the electric field intensity of 5 kV/cm,120 pulses,extraction time of 30 min,and 2%material concentration.The PEFcontained more polyphenols than the conventional water extraction and ultrasound-assisted extraction.The Phyllanthus emblica extract had substantial antioxidant and anti-inflammatory effects,with a good clearance effect on DPPH(IC50 of 1.82%)and ABTS(IC50 of 1.80%)radicals.At the Phyllanthus emblica extract concentration of 1.25%,inflammatory factors(TNF-α)were reduced by 47.08%;and Conclusion:The PEF is a leading-edge and promising method for preparing Phyllanthus emblica extracts.

Network pharmacology and experimental validation to reveal the anti-hepatitis B virus pharmacological mechanism of Phyllanthus urinaria L.

Background:To explore the pharmacological mechanism of the anti-hepatitis B virus of Phyllanthus urinaria L.through network pharmacological analysis and experimental validation.Method:The active ingredient,target of action and target of action related to hepatitis B were clarified by searching the herb group identification,GeneCards and OMIM databases,and the protein interaction relationship was obtained by using the String database,and the protein interaction network map was constructed by using Cytoscape software.We also performed gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of key targets of the anti-hepatitis B action of Phyllanthus urinaria L.and predicted the core targets and pathways of Phyllanthus urinaria L.anti-hepatitis B.The main targets predicted by network pharmacology were then validated by HepG2.2.15 cell experiments.Results:By searching active ingredient targets and hepatitis B disease targets,a total of 19 active ingredients and 64 related targets of action were retrieved from Phyllanthus urinaria L.,and a total of 51 common targets were obtained by mapping the obtained hepatitis B disease targets and drug targets.protein protein interaction network analysis indicated that targets including TNF,JUN,AKT1,IL-10,IL-1B,CAT,HMOX1,NFE2L2,and CASP3 and other targets may be the core targets.gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that the treatment of hepatitis B by Phyllanthus urinaria L.mainly included inflammation and oxidation-related processes,and the signaling pathways mainly included fluid shear stress and atherosclerosis,VEGF,and hepatocellular carcinoma.The results of the in vitro test showed that after the action of different concentrations of the extracts of the Phyllanthus urinaria L.in the safe concentration range on cells HepG2.2.15,HBsAg,HBeAg and hepatitis B virus DNA levels were significantly inhibited,and NFE2L2 and HMOX1 were affecting hepatitis B virus transcription and replication by regulating the oxidative stress response.Conclusion:Using an integrated network pharmacology approach,this study revealed the active components and potential targets of Phyllanthus urinaria L.for the treatment of the hepatitis B virus,providing a theoretical basis for the research and clinical application of Phyllanthus urinaria L..

Suppressed effects of Phyllanthus urinaria L.ethyl acetate extract on hepatitis B virus both in vitro and in vivo

Background:Phyllanthus urinaria L.(P.urinaria)extract(PUE)has been used to inhibit hepatitis B virus(HBV).However,the underlying mechanism remains unclear.To investigate which PUE fractions and main components lead to against HBV and approach the relevant molecular mechanisms.Methods:P.urinaria was extracted with water,and then the decoction was extracted by petroleum ether,ethyl acetate,and n-butanol in turn.The HepG2.2.15 cell was treated with aqueous fraction,petroleum ether fraction,ethyl acetate fraction and n-butanol fraction,gallic acid(GA,C7H6O5)and corilagin(CL,C27H22O18),respectively.The medium was collected for hepatitis B surface antigen(HBsAg)and hepatitis B e antigen assays.Cell counting kit-8 method was used to identify cell proliferation.Also,the levels of cellular oxygen consumption,reactive oxygen species,and reduced glutathione were detected.The HBV modeling mice were treated with ethyl acetate fraction,entecavir and physiological saline,respectively.The serum was collected for HBsAg and inflammatory cytokines assays.Liver tissue metabolites were screened by LC-MS/MS method.Results:The ethyl acetate fraction(EAF)of P.urinaria could significantly inhibit HBV secretion in HepG2.2.15(P<0.05).Furthermore,two main constitutes in ethyl acetate fraction,GA and CL,could significantly inhibit HBV secretion and reduced cell proliferation(P<0.05).Also,GA and CL could increase cellular oxygen consumption,intracellular superoxide anions level,superoxide dismutase level and glutathione depletion.Compared with the Modeling group,EAF significantly decreased the expression levels of HBsAg,IL-1β,IFN-α(P<0.05).LC-MS/MS analysis results showed that EAF dramatically up-regulate hydroxyproline,maltotriose,betaine and down-regulate glutathione disulfide,taurocholate,taurochenodeoxycholate(P<0.05).Kyoto Encyclopedia of Genes and Genomes results show that the differential metabolites were mainly enriched in ATP-binding cassette transporters pathway.Conclusions:P.urinaria exhibits suppressed effects on HBV by modulating reactive oxygen species formation or metabolomics both in vitro and in vivo.These data indicate that P.urinaria may be an alternative therapeutic agent for the treatment of HBV-related hepatitis.

Effect of Phyllanthus emblica Linn.on Candida adhesion to oral epithelium and denture acrylic

Objective:To investigate the effect of Phyllanthus emblica(P.emblica) Linn,ethanolic extract on the adhesion of Candida albicans(C.albicans) to human buccal epithelial cells(BECs) and denture acrylic surfaces.Methods:Human BECs and transparent acrylic strips were pretreated with ethanolic extract solution of P.emblica fruits at concentration ranged from 18.7 to 300 mg/mL.After washing BECs and the strips were inoculated with three strains of C.albicans (ATCC 10281 and two clinical isolates)(10~7 cells/mL).Normal saline solution(NSS) and 0.2% chlorhexidine gluconate were used as negative and positive controls,respectively.BECs were harvested on 12μm-polycarbonate filters(Millipore,USA).The membrane filters and the strips were stained with Gram stain.Adherent yeast cells on 100 randomly selected epithelial cells and 20 randomly selected fields on each strip were counted under microscope.The statistical significance was calculated by Kruskal-Wallis and Tukey tests at a significant level of P【0.05. Results:Significant lower numbers of all strains of yeasts adhering to BECs and acrylic strips were observed after exposure to 75-300 mg/mL of plant extract compared with NSS.Conclusions: The present study demonstrates that P.emblica ethanolic extract interferes with the adhesion of C. albicans to BECs and denture acrylic surfaces in vitro.

Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents:Doxorubicin and cisplatin against human hepatocellular carcinoma and lung cancer cells

AIM: To examine the growth inhibitory effects of Phyllanthus emblica (P. emblica) and Terminalia bellerica (T. bellerica) extracts on human hepatocellular carcinoma (HepG2), and lung carcinoma (A549) cells and their synergistic effect with doxorubicin or cisplatin. METHODS: HepG2 and A549 cells were treated with P. emblica and T. bellerica extracts either alone or in combination with doxorubicin or cisplatin and effects on cell growth were determined using the sulforhodamine B (SRB) assay. The isobologram and combination index (CI) method of Chou-Talalay were used to evaluate interactions between plant extracts and drugs. RESULTS: P. emblica and T. bellerica extracts demonstrated growth inhibitory activity, with a certain degree of selectivity against the two cancer cell lines tested. Synergistic effects (CI < 1) for P. emblica /doxorubicin or cisplatin at different dose levels were demonstrated in A549 and HepG2 cells. The T. bellerica/ cisplatin or doxorubicin also showed synergistic effects in A549 and HepG2 cells. In some instances, the combinations resulted in antagonistic effects. The dose reduction level was different and specific to each combination and cell line. CONCLUSION: The growth inhibitory activity of doxorubicin or cisplatin, as a single agent, may be modified by combinations of P. emblica or T. bellerica extracts and be synergistically enhanced in some cases. Depending on the combination ratio, the doses for each drug for a given degree of effect in the combination may be reduced. The mechanisms involved in this interaction between chemotherapeutic drugs and plant extracts remain unclear and should be further evaluated.

Efficacy and safety of Amla(Phyllanthus emblica L.) in non-erosive reflux disease:a double-blind,randomized,placebo-controlled clinical trial

Background： Gastroesophageal reflux disease （GERD） is one of the most common gastrointestinal com- plaints. GERD, caused by the reflux of stomach contents into the esophagus, leads to troublesome symptoms such as heartburn and regurgitation. It is classified into two types： erosive esophagitis, characterized by visible esophageal mucosa erosion in endoscopy, and non-erosive reflux disease （NERD）. GERD is a chronic and recurrent disease that impairs the quality of life and imposes socioeco- nomic and therapeutic burdens to both patients and society. Objective： Due to the failure of the conventional treatments for GERD and to the traditional use of Amla （Phyllanthus emblica L.）, in addition to beneficial effects shown in recent studies, we evaluated the safety and efficacy of Amla tablet for improvement of symptoms of patients with NERD. Design, setting, participants and interventions： We designed a double-arm, randomized, double-blind, placebo-controlled clinical trial. Sixty-eight patients who had classic symptoms of GERD （heartburn, regurgitation and epigastralgia） for at least three months before the start of the trial were randomized in two parallel groups. Patients in the Amla group received two 500 mg Amla tablets twice a day, after meals, for 4 weeks. In the control group, patients received placebo tablets similar to the Amla prescription. Main outcome measures： The patients were visited at baseline, and at the end of the 2nd and 4th weeks of intervention; their symptoms were measured on a frequency and severity scale for the symptoms of NERD, according to the quality of life in reflux-associated disease questionnaire. Results： Frequencies of heartburn and regurgitation in both groups of the study were significantly reduced after intervention （P 〈 0.001）. Repeated measures logistic regression analysis showed that, in the Amla group, there was a more significant reduction in regurgitation frequency, heartburn frequency, regurgita- tion severity and heartburn severity during the study period, compared with the placebo group （P 〈 0.001 ）. Conclusion： This randomized double-blind, placebo-controlled clinical trial demonstrated that Amla could reduce frequencies of heartburn and regurgitation and improve heartburn and regurgitation severity in patients with NERD.

Investigation of Antidiabetic Effect of Ethanolic Extract of <i>Phyllanthus emblica</i>Linn. Fruits in Experimental Animal Models

Hypoglycemic effects were investigated in the ethanolic extract of fruits of Phyllanthus emblica Linn. with long evans rats (100-120 g). Residual gut sucrose content, gut perfusion and disaccharidase activity studies were carried out to assess these activities. The residual gut sucrose content study was carried out to reveal the amount of sucrose remaining in the gastro intestinal tract (GIT) at six different segments. The ethanolic fruit extract of Phyllanthus emblica Linn. significantly increases the residual sucrose content throughout the gut after sucrose ingestion. Study on intestinal enzyme activity showed that the extract caused a significant (p 0.05), dose dependent inhibition of intestinal disaccharidase enzyme activity. Evaluation of the effect of the plant extract on intestinal enzyme activity showed significant inhibition of intestinal disaccharidase activity, suggesting that reduction of sucrose absorption may be partly related to the inhibition of disaccharidase activity in the gut. The gut perfusion analysis showed that the extract significantly reduced intestinal glucose absorption. The results of the present study suggest that ethanolic extract of Phyllanthus emblica Linn. has significant antidiabetic effects in a dose dependent manner and this may be effective in the treatment of diabetes.

HPLC指纹图谱结合4种指标含量测定的余甘子质量评价研究

建立余甘子(Phyllanthus emblica L.)HPLC指纹图谱,并对4种化学成分的含量进行测定,为余甘子是否去核使用提供了一定参考。采用CAPCELL PAK C_(18)色谱柱,以乙腈-0.1%磷酸水溶液作为流动相,检测波长273 nm,流速为1.0 mL/min,柱温为30℃,进样量10μL,建立了云南32个不同产地余甘子的指纹图谱。通过对照品比对并结合光谱分析,对共有峰进行鉴定;借助中药色谱指纹图谱相似度评价系统对32批产地的余甘子的图谱进行相似度评价,通过聚类分析(HCA)、主成分分析(PCA)比较两种规格余甘子的差异,寻找差异性成分;并对两种规格余甘子药材4种有效成分含量进行对比研究。32批余甘子指纹图谱中有17个共有峰,指认出其中4个成分,相似度分析均大于0.90,对其进行聚类分析,有3个主成分,可聚为6类。对没食子酸、鞣花酸、柯里拉京、诃子酸定量分析,4个化学成分在各自范围内线性良好(r≥0.9999),精密度、重复性、稳定性试验均符合定量要求,加标回收率为98.77%~100.92%,RSD均小于2.0%(n=9)。含量测定结果显示,有核余甘子中没食子酸、柯里拉京、鞣花酸的含量均高于无核余甘子,而无核余甘子中诃子酸高于有核余甘子。所建立的方法简便、稳定、可靠,HPLC指纹图谱结合多指标成分含量测定可用于余甘子药材质量评价,为余甘子是否去核使用,提供了一定参考。

Prolonged Seed Dormancy in <i>Phyllanthus emblica</i>L. Can Be Overturned by Seed Scarification and Gibberellin Pre Treatment

Phyllanthus emblica L. is an important constituent of Ayurvedic medicine and a fresh fruit species in the market in Sri Lanka. Therefore, it has a high potential to be established as a commercial fruit crop. The seeds of P. emblica are semi orthodox and exhibit a long dormancy period hindering the natural sexual propagation. Therefore, it still remains as an underutilized fruit crop in Sri Lanka due to its lack of quality planting material and poor propagation techniques. Long term dormancy also causes a big challenge in germinating seeds to create populations in breeding programs. In order to promote P. emblica from its underutilized status, what seems most feasible is to develop a method to break up the seed dormancy artificially. In order to do so, the methods of breaking the dormancy of P. emblica seeds have to be studied. Therefore, the aim of this study was to identify a method to break up the seed dormancy of P. emblica. The seeds were extracted from 21 trees belonging to three different districts in Sri Lanka. The selected viable seeds were subjected to four different pre treatments: none treated seeds (i.e. control), seeds scarified, seeds scarified and treated with 1% gibberellin and seed coat removed and followed by seeds treated with 1% gibberellin. From the four treatments, seed dormancy was overturned with a germination percentage of 43% by the seed pre treatment where the seeds were scarified and treated with 1% gibberellin and no other pre treatment methods were successful in breaking the dormancy. This suggests that the natural germination potential of P. emblica seeds is very low and it can be overridden by seed scarification and gibberellin pre treatment.

Investigation of the Phytoconstituents and Bioactivity of Various Parts of Wild Type and Cultivated <i>Phyllanthus emblica</i>L.

Phyllanthus emblica L. belonging to the family Euphorbiaceae is a common medicinal plant in Bangladesh. In order to evaluate the phytoconstituents and bioactivity of various parts of P. emblica, both the wild type (i.e., small) and cultivated (i.e., big) fruits were collected from the local market of Bangladesh and six ethanolic extracts were prepared from various parts of the collected fruits for this current study. A comparative analysis of the phytochemical compositions and various bioactivities especially antibacterial, antifungal, and antioxidant activity of the six ethanolic extracts were accomplished. The qualitative phytochemical analysis of plant extracts revealed the presence of large amounts of proteins, carbohydrates, phenolic compounds, glycosides, alkaloids, coumarins, flavonoids, tannins, saponins and resins etc. The maximum antioxidant activity was observed for wild type P. emblica seed extracts and the minimum activity was observed for cultivated flesh extracts. Moreover, among the samples the wild type P. emblica extracts have excellent amount of total phenol contents and the highest free radical scavenging activity. It is also found that P. emblica samples were active against both gram-positive (i.e., B. subtilis) and gram-negative (i.e., E. coli, and S. typhi) bacteria. The highest antibacterial activity was achieved in the wild type P. emblica seed extracts against both pathogenic (S. typhi) and non-pathogenic bacterial strains (E. coli, and B. subtilis). These samples showed almost no activity against Fusarium sp. fungus.

基于SNP和InDel标记的余甘子群体遗传分析

【目的】采用高通量测序技术解析余甘子种质资源的群体遗传结构和遗传多样性,为余甘子系统分类、遗传资源创新利用提供理论基础。【方法】利用ddRADseq技术对112份余甘子种质资源进行高通量简化基因组测序,利用Cutadapt和Trimmomatic软件对原始数据进行过滤,筛选得到高质量测序数据;使用MUNEAK软件进行多态性标记发掘,基于获得的SNP和InDel标记,进行群体结构分析、主成分分析、系统发育分析及遗传多样性分析。【结果】余甘子测序样品共获得8934个SNP和InDel标记,群体结构分析将余甘子种质分为2个类群,类群划分与种质来源地相关,该结果与主成分分析和系统发育分析相一致。余甘子各种质间遗传距离为0.027~0.459,平均遗传距离为0.248;云南地区的余甘子种质的期望杂合度、观测杂合度及多态性信息含量值最高,依次为0.267、0.184及0.218;余甘子群体间的Fst在0.080~0.266之间,群体遗传分化程度中等偏高。【结论】该测序技术可有效地解析余甘子种质的群体结构和遗传多样性,为余甘子种质资源的鉴定评价、系统分类及遗传多样性研究提供参考。

Effects of 1,2,4,6-tetra-O-galloyl-β-D-glucose from P. emblica on HBsAg and HBeAg Secretion in HepG2.2.15 Cell Culture

A polyphenolic compound, 1,2,4,6-tetra-O-galloyl-β-D-glucose （1246TGG）, was isolated from the traditional Chinese medicine Phyllanthus emblica L. （Euphorbiaceae） and assayed for its potential as an anti-hepatitis B virus （HBV） agent. The cytotoxicity of 1246TGG on HepG2.2.15 as well as HepG2 cells was determined by observing cytopathic effects, and the effects of 1246TGG on secretion of HBsAg and HBeAg in HepG2.2.15 cells were assayed by enzyme immunoassay. Results indicates that treatment with 1246TGG （6.25 ~tg/mL, 3.13 ~tg/mL）, reduced both HBsAg and HBeAg levels in culture supernatant, yet the inhibitory effects tend to decline with the assay time. This study provides a basis for further investigation of the anti-HBV activity and possible mechanism of action of 1246TGG

Phyllanthus acidus(L.) Skeels and Rhinacanthus nasutus(L.) Kurz leaf extracts suppress melanogenesis in normal human epidermal melanocytes and reconstitutive skin culture

Objective: To determine the effect of extracts from Phyllanthus acidus(P. acidus)(L.) Skeels and Rhinacanthus nasutus(R. nasutus)(L.) Kurz leaves on melanogenesis and the underlying mechanism in normal human epidermal melanocytes(NHEM) and a reconstitutive skin model. Methods: NHEM and a reconstitutive skin model were stimulated with ethanol extracts of P. acidus(L.) Skeels and R. nasutus(L.) Kurz leaves. mRNA expression of microphthalmiaassociated transcription factor(MITF), tyrosinase(TYR), tyrosinase-related protein 1(TYRP1) and dopachrome tautomerase(DCT) were examined by real-time PCR. The melanin content in NHEM was also measured. Moreover, protein levels of tyrosinase were determined using western blot analysis.Results: In NHEM and the reconstitutive skin model, ethanol extracts from P. acidus(at 12.5 and 25.0 μg/mL) and R. nasutus(at 6.25 and 12.50 μg/mL) significantly diminished mRNA expression of MITF, TYR, TYRP1 and DCT in a concentration-dependent manner. P. acidus and R. nasutus extracts also reduced the amount of melanin in α-MSH-stimulated NHEM. Moreover, P. acidus and R. nasutus extracts markedly suppressed tyrosinase at the translational level in the reconstitutive skin model. Conclusions: P. acidus and R. nasutus extracts significantly reduced melanogenesis in NHEM and the reconstitutive skin model, suggesting that P. acidus and R. nasutus extracts can inhibit melanin synthesis through downregulation of MITF, TYR, TYRP1 and DCT. Therefore, the ethanol extracts of P. acidus and R. nasutus contain compounds that have the potential for development as a skin lightening agent for the treatment of hyperpigmentation disorder or melasma.

苗药吉祥草配伍余甘子抗Lewis肺癌细胞增殖组分的筛选

目的:探索吉祥草和余甘子配伍后治疗非小细胞肺癌的药效物质,为吉祥草配伍余甘子在抗肿瘤制剂开发方面的研究提供实验依据。方法:采用鲜药榨汁、传统水煎、渗漉为提取方法制得吉祥草配伍余甘子的不同提取物,以Lewis肺癌细胞体外实验模型作为评价手段,优选药材的最佳提取方法;通过膜分离技术,将最佳提取方法的提取物分段截留得到不同分子量区间段组分,以Lewis肺癌细胞体外实验模型作为评价手段优选最佳的分子量区间段。结果:吉祥草配伍余甘子对Lewis肺癌细胞增殖抑制作用的最佳提取方法为95%乙醇渗漉法。95%乙醇渗漉提取物对Lewis肺癌细胞增殖抑制作用的最佳分子量区间段组分为分子量10~30 KD与分子量≤1 KD组分。结论:苗药吉祥草配伍余甘子对Lewis肺癌细胞具有增殖抑制作用,抑制作用最佳组分为95%乙醇渗漉的分子量10~30 KD与分子量≤1 KD组分。