In vitro bioactivity-guided fractionation and characterization of polyphenolic inhibitory fractions from Acacia nilotica (L.) Willd. ex Del

The present study was undertaken to evaluate antimutagenic and cytotoxic effects of different extracts/fractions of Acacia nilotica prepared by maceration method. The potency order of different extracts was more or less similar in Ames assay as well as in cytotoxic assay. Considering the maximum potential of acetone extract in both the assays, the studies were initiated to fractionate this extract. Two pure fractions, namely AN-1 and AN-2, were obtained from acetone extract, of which AN-2 was found to be of gallic acid and AN-1 fraction is still to be identified. In conclusion, the antimutagenic and cytotoxic activities exhibited by acetone extract may partially be ascribed to the presence of gallic acid and other polyphenols.     

Potential cytotoxic activity of some Brazilian seaweeds on human melanoma cells

In vitro screening of the crude extracts of some Brazilian coastal seaweeds for cytotoxic activity against a cultured human melanoma cancer cell line using the sulphorhodamine B assay was performed. The crude dichloromethane:chloroform extract of Stypopodium zonale showed good cytotoxic activity against the C32 cell line. The crude acetone extract and aqueous phase of Lobophora variegata did not show any activity, but semi-purified fractions XAD LOB I and II could inhibit the growth of melanoma cells. The crude acetone extract of Caulerpa racemosa showed some cytotoxicity, but caulerpin isolated from this extract did not show any such activity. The crude acetone extract of Spatoglossum schroederi was not able to inhibit the growth of C32 melanoma cells.     

Crown gall -- a plant tumour with biological activities

Petroleum ether, acetone, 80% MeOH and water extracts of crown gall, a plant tumour, obtained from Eucalyptus globulus tree were screened for cytotoxic, antioxidant, antiinflammatory, embryotoxic, antitumour-promoting and antimicrobial activities.In terms of bioactivity the 80% MeOH extract was most effective followed by the acetone extract. The petroleum ether extract showed weak to moderate cytotoxic activity in dose-dependent manner against PC12 cells, mouse L fibroblasts and 1321N1 glia cells, whereas the hydroalcohol extract had no or a weak cytotoxic effect. The 80% MeOH extract exhibited strong antioxidant activity. Based on the in vitro HET-CAM assay all the extracts were effective against inflammation. The extracts did not show any embryotoxic effect at the concentrations tested. Antitumour-promoting activity (100% inhibition; 100 microg/mL) was observed in the 80% MeOH and acetone extracts. In the antimicrobial screening all extracts displayed predominantly antifungal activity against Candida sp. The extracts also showed various levels of antibacterial activity against E. faecalis, Ps. aeruginosa, Bac. subtilis and Staph. epidermidis.From the results of the investigations it can be concluded that crown gall is a valuable plant tumour tissue having interesting biological activities.     

The extract of Hibiscus syriacus inducing apoptosis by activating p53 and AIF in human lung cancer cells

Natural products including plants, microorganisms and marine life provide rich resources for anticancer drug discovery. The root bark of Hibiscus syriacus has been used as an antipyretic, anthelmintic and antifungal agent in Asia. The antiproliferative effects of H. syriacus on human lung cancer cells were evaluated with bio-assays. The apoptotic activity was detected by Hoechst 33342 DNA staining and annexin V staining. The expression of caspases, p53, apoptosis induced factor (AIF), Bcl-2 and Bax were evaluated with Western blotting. The in vivo anticancer activity was evaluated using A549-xenograft model. The acetone extract of H. syriacus (HS-AE) exhibited a better cytotoxic effect on lung cancer cells than its methanol extract (HS-ME) or water extract (HS-WE). The IC(50) values of HS-AE on A549 (adenocarcinoma), H209 (squamous cell carcinoma) or H661 (large cell carcinoma) lung cancer cells ranged from 14 to 22 microg/ml after 48 hours of treatment. After 48 hours of exposure, HS-AE (15 microg/ml) induced A549 cell apoptosis to 48 +/- 3.6% of the control. Using Western blotting, HS-AE appears to suppress the expression of p53 and AIF. The results of the in vivo study showed that HS-AE suppresses growth in A549 subcutaneous xenograft tumors. These results indicate that HS-AE exerts significant and dose-dependent antiproliferative effects on cancer cells in vitro and in vivo, which prompts us to further evaluate and elucidate the bioactive component(s) of H. syriacus.     

Biological activity of barbados cherry (acerola fruits, fruit of Malpighia emarginata DC) extracts and fractions

Fractionation of barbados cherry (acerola fruit, a fruit of Malpighia emarginata DC.) extracts were performed by organic solvent extractions and column chromatographies, using two extraction methods. Higher cytotoxic activity was concentrated in fractions A4 and A6 (acetone extract), and H3 and HE3 (hexane extract). These four fractions showed higher cytotoxic activity against tumor cell lines such as human oral squamous cell carcinoma (HSC-2) and human submandibular gland carcinoma (HSG), when compared with that against normal cells such as human periodontal ligament fibroblasts (HPLF) and human gingival fibroblasts (HGF). HE2 (hexane extract), AE2 (ethyl acetate extract), AE3, AE4, AE5, A8, A9 and A10 showed some relatively higher anti-bacterial activity on the Gram-positive Staphylococcus epidermidis ATCC 1228 but were ineffective on the representative Gram-negative species E. coli and Ps. aeruginosa. The fractions were inactive against Helicobacter pylori, two representative Candida species, and human immunodeficiency virus (HIV). H3, H4 and HE3, which displayed higher tumor-specific cytotoxicity also showed higher multidrug resistance (MDR) reversal activity, than (+/-)-verapamil as positive control. ESR spectroscopy shows that the radical-mediated oxidation is not involved in the induction of tumor-specific cytotoxic activity. The tumor specific cytotoxic activity and MDR reversal activity of barbados cherry may suggest its possible application for cancer therapy.     

基于UPLC-Q-Orbitrap-MS整合网络药理学研究藏药大三果化学成分及作用机制＊

目的 利用UPLC-Q-Orbitrap-MS技术系统研究藏药复方大三果主要化学成分并结合网络药理学探讨发挥药效的主要作用机制。方法 采用UPLC-Q-Orbitrap-MS对大三果主要化学成分进行分析，根据化合物的一级、二级质谱信息，结合相关文献和对照品，对化学信息进行快速识别。采用网络药理学的方法对大三果的成分靶点构建化合物-靶点网络，通过蛋白相互作用（PPI）网络筛选出核心靶点，对靶点进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析。结果 从大三果醇提物中共鉴别出85个学成分，主要包括酚酸类、鞣质类、黄酮类成分。根据ADME过筛标准获得12个主要活性成分，包括Quercetin，Kaempferol，Luteolin，Morin和Ellagic acid等作用于10个关键靶点，包括CXCL8、APP、CHRM2、CXCL2、CXCL10、ADCY2、CXCR1、CXCL11、PTGER3、ADORA1等，富集的信号通路主要集中在对炎症、癌症、免疫等方面。结论 本研究从网络药理学角度阐明大三果多成分、多靶点、多途径的整体调节特点，初步揭示了抗炎、抗癌、免疫调节的物质基础和作用机制，为大三果进一步的临床应用和深入研究提供了思路和线索。     

Kaempferitrin induces apoptosis via intrinsic pathway in HeLa cells and exerts antitumor effects

Ethnopharmacological relevance

Justicia spicigera is used for the empirical treatment of cervical cancer in Mexico. Recently, we showed that Justicia spicigera extracts exerted cytotoxic and antitumoral effects and the major component of this extract was kaempferitrin (KM).

Materials and methods

The cytotoxic and apoptotic effect of KM on human cancer cells and human nontumorigenic cells were evaluated using MTT and TUNEL assays, and Annexin V/Propidium iodide detection by flow cytometry. The effect of KM on cell cycle was analyzed by flow cytometry with propidium iodide. The apoptotic and cell cycle effects were also evaluated by western blot analysis. Also, different doses of KM were injected intraperitoneally daily into athymic mice bearing tumors of HeLa cells during 32 days. The growth and weight of tumors were measured.

Results

KM induces high cytotoxic effects in vitro and in vivo against HeLa cells. The general mechanisms by which KM induces cytotoxic effects include: cell cycle arrest in G1 phase and apoptosis via intrinsic pathway in a caspase dependent pathway. Also, KM exerts chemopreventive and antitumor effects.

Conclusion

KM exerts cytotoxic and antitumor effects against HeLa cells.     

Inhibition of fowlpox virus by an aqueous acetone extract from galls of Guiera senegalensis J. F. Gmel (Combretaceae)

An aqueous acetone extract from the galls of Guiera senegalensis was screened for in vitro antiviral activity against fowlpox virus (FPV). Cytopathic effect (CPE) inhibition and plaque inhibition assays were used to show presence of antiviral effects against FPV, whilst cytotoxicity assays established the relative safety of the extract for cells in vitro. Phytochemical analyses revealed the presence of phenolic compounds including flavonoids, tannins and anthocyanins as well as steroids and alkaloids. Thin-layer chromatographical (TLC) analysis also revealed the presence of quercitrin, quercetin, kaempferol, apigenin, rutin, gallic acid as well as unknown flavonoids and unknown phenolic acids. The antiviral effect of the extract was partially attributed to phenolic components including flavonoids.     

Aqueous extract of Phyllanthus urinaria induces apoptosis in human cancer cells

Cell apoptosis is now known to play an important role in the maintenance of cellular homeostasis and anti-carcinogenesis. The anticancer effect of aqueous extract prepared from Phyllanthus urinaria (P. urinaria) was investigated by analyzing its potential to induce apoptosis in human cancer cells. We showed that the aqueous extract of P. urinaria could reduce the viability by inducing the apoptosis in human cancer cells derived from several different origins as demonstrated by morphological changes and DNA fragmentation. Yet, P. urinaria extract exhibited no cytotoxic effect on normal human cells, including vascular endothelial cells and liver cells under the same conditions. It suggests that the aqueous extract of P. urinaria is substantially useful in treating various kinds of human cancer cells without toxic side effect on normal cells.     

Rhizoma Paridis ethanol extract selectively inhibits the proliferation of HUVECs comparing to Lovo cells and shows anti-angiogenesis effects in a mouse model

Ethnopharmacological relevance

Rhizoma Paridis, a traditional Chinese Medicine, was identified to be cytotoxic to cancer cells. The present study was designed to investigate the potential anti-angiogenic and antitumor effect of the ethanol extract of Rhizoma Paridis (RPE) in vitro and in vivo.

Materials and methods

The cytotoxic effect of RPE against human colon cancer Lovo cells and human umbilical vein endothelial cells (HUVECs) was examined using MTT assay. We also tested the effect of RPE on tube formation, migration, apoptosis and cell cycle of HUVECs. Moreover, Lovo subcutaneous xenograft was applied to study the antitumor and anti-angiogenesis effect of RPE in vivo.

Results

RPE exerted a higher inhibition effect on the proliferation of HUVECs than Lovo cells. The tube formation and cell migration were also significantly inhibited in the presence of RPE in a concentration-dependent manner though the significant inhibition effects were observed at the cytotoxic dose. RPE induced cell apoptosis and G0–G1 cell cycle arrest of HUVECs. In vivo, significant tumor growth inhibition was observed in human colon cancer xenografts established by Lovo cells, accompanying by a marked decrease in MVD.

Conclusions

Our current study exhibited that RPE has a selective cytotoxity against HUVECs comparing to Lovo cells and also demonstrated significant anti-angiogenic effect in vivo.     

Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. fruit

A 70% methanol extract of Terminalia chebula fruit, was studied for its effects on growth in several malignant cell lines including a human (MCF-7) and mouse (S115) breast cancer cell line, a human osteosarcoma cell line (HOS-1), a human prostate cancer cell line (PC-3) and a non-tumorigenic, immortalized human prostate cell line (PNT1A) using assays for proliferation ([(3)H]-thymidine incorporation and coulter counting), cell viability (ATP determination) and cell death (flow cytometry and Hoechst DNA staining). In all cell lines studied, the extract decreased cell viability, inhibited cell proliferation, and induced cell death in a dose dependent manner. Flow cytometry and other analyses showed that some apoptosis was induced by the extract at lower concentrations, but at higher concentrations, necrosis was the major mechanism of cell death. ATP assay guided chromatographic fractionation of the extract yielded ellagic acid, 2,4-chebulyl-beta-D-glucopyranose (a new natural product), and chebulinic acid which were tested by ATP assay on HOS-1 cell line in comparison to three known antigrowth phenolics of Terminalia, gallic acid, ethyl gallate, luteolin, and tannic acid. Chebulinic acid (IC(50) = 53.2 microM +/- 0.16) > tannic acid (IC(50) = 59.0 microg/ml +/- 0.19) > and ellagic acid (IC(50) = 78.5 microM +/- 0.24), were the most growth inhibitory phenolics of T. chebula fruit in our study.     

Extracts from the roots of Lindera strychifolia induces apoptosis in lung cancer cells and prolongs survival of tumor-bearing mice

Lindera strychifolia, a scandent shrub Lauraceous medicinal plant, has been used in Chinese traditional medicine as a palliative and an anti-spasmodic. It also shows cytotoxic effects against several tumor cell lines and inhibits marcromolecule biosynthesis. This study investigated the anti-tumor effects of L. strychifolia extract against lung cancer cells using in vitro and in vivo models. Two human lung cancer cell lines A549 (adenocarcinoma) and SBC-3 (small cell carcinoma), and a non-tumor cell line 3T3-L1 (mice fibroblasts) were subjected to L. strychifolia extract treatment. On lung cancer cells, L. strychifolia induced cell growth inhibition in a dose-dependent manner. Conversely, the extract did not show any significant cytotoxic effect on 3T3-L1 cells. Therefore, the extract is specific for tumor cells. Tumor cells treated with L. strychifolia extract showed typical morphological appearance of apoptosis including nuclei fragmentation and cell condensation. The in vivo effects of L. strychifolia extract were investigated in C57BL/6 mice transplanted with Lewis lung cancer (LL-2) cells, and in BALB/c nude mice transplanted with A549 or SBC-3 human lung cancer cells. Oral administration of L. strychifolia extract prolonged survival time and inhibited tumor growth in a dose-dependent manner by inducing apoptosis in the LL-2 cell mice model. Furthermore, in A549 or SBC-3 cell nude mice models, oral administration of L. strychifolia extract also significantly inhibited tumor growth at the 5.0 mg/ml concentration. These findings suggested that the components of L. strychifolia have anticancer activity and may contribute to clinical applications in the prevention and treatment of lung cancer.     

Anti-metastatic effects of aqueous extract of Helixanthera parasitica

Metastasis, the spread of cancer in body, is a major cause of death. We have screened anti-metastatic activity of aqueous and dichloromethane extracts of several not previously studied Thai herbs, using an in vitro invasion test. This involves the in vitro invasion of HCC-S102, a hepatocellular carcinoma cell line derived from a Thai patient, through a reconstituted-basement membrane (Matrigel). The aqueous extract of a plant (Helixanthera parasitica) revealed a significant inhibitory effect on the cancer cell invasion, and showed antioxidant activity. The aqueous extract was partially purified by silica gel column chromatography, and the highest anti-metastatic activity fraction showed 83% inhibition of invasion with low cytotoxic effect. However, anti-metastatic activity was not associated with the antioxidant activity of the aqueous extract.     

Cytotoxic effects of Echinacea purpurea flower extracts and cichoric acid on human colon cancer cells through induction of apoptosis

Ethnopharmacological relevance

Echinacea is a top-selling herbal supplement that acts as immunostimulant. It has been used to treat common cold, coughs, bronchitis and upper respiratory infections. It is also a popular product used in anticancer therapy. The cytotoxic effects of Echinacea on cancer cells are still not clear. The aims of this study were to provide a preliminary validation of the effects of 50% aqueous ethanol extract of Echinacea purpurea flowers and its major compound, cichoric acid, on human colon cancer cells Caco-2 and HCT-116.

Materials and methods

The cytotoxic effects of Echinace flower extracts and cichoric acid on cell viability, telomerase activity, DNA fragmentation, β-catenin, caspase-9, and cleavage of poly-ADP-ribose polymerase (PARP) of human colon cancer cell were examined.

Results

The results showed a significant inhibition of proliferation in E. purpurea flower extract and cichoric acid in a dose- and time-dependent manner. Cichoric acid treatment decreased telomerase activity in HCT-116 cells. Moreover, cichoric acid effectively induced apoptosis in colon cancer cells, which were characterized by DNA fragmentation, activation of caspase-9, cleavage of PARP and downregulation of β-catenin.

Conclusions

Our data indicate that cichoric acid has a strong growth-inhibitory effect against colon cancer cells, presumably resulting from the reduced telomerase activity and the induction of apoptosis. The exact mechanism of action should still be determined in future studies. Overall, the effects of 50% aqueous ethanol extract of E. purpurea flowers and cichoric acid may have provided in vitro evidence for the use as chemotherapeutic agents.     

枇杷叶紫珠化学成分及其细胞毒活性和抗炎活性

目的研究枇杷叶紫珠Callicarpa kochiana的化学成分及其细胞毒活性和抗炎活性。方法枇杷叶紫珠90%丙酮提取物采用硅胶、Sephadex LH-20、MCI、ODS、重结晶、半制备HPLC进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。采用CCK8法、Griess法、ELISA法测定化合物4~5的细胞毒活性和抗炎活性。结果从中分离得到5个化合物,分别鉴定为3,7,3’,4’-tetramethyl-quercetin (1)、pachypodol (2)、14α-hydroxyisopimaric acid(3)、14α-hydroxy-7,15-isopimaradien-18-oic acid (4)、(16R)-16,17-dihydroxyphyllolladan-3-one (5)。化合物4在一定浓度范围内对7种肿瘤细胞具有不同程度的抑制作用,并可以减少RAW264. 7细胞NO、IL-10、MCP-1、TNF-α的释放量。结论化合物1~5均为首次从该植物中分离得到,化合物4具有细胞毒活性和抗炎活性。     

The cytotoxicity and chemical constituents of the hexane fraction of Typhonium flagelliforme (Araceace)

The plant, Typhonium flagelliforme (Araceae), commonly known as the "rodent tuber" in Malaysia, is often used as an essential ingredient of herbal remedies for alternative cancer therapies. The hexane extract of this plant was evaluated for cytotoxic activity against in vitro culture on P388 murine leukaemia cells and showed weak IC(50) of 15 microg/ml. The partial chemical constituents were identified as methyl esters of hexadecanoic acid, octadecanoic acid, 9-octadecenoic acid and 9,12-octadecadienoic acid. In addition, several common aliphatics were identified as dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane and eicosane. The unique methyl ester of 13-phenyltridecanoic acid was isolated and positively identified using spectroscopic methods. None of the identified compounds showed or are known to have cytotoxic behaviour.     

不同溶剂对鸡血藤提取物总黄酮含量 及抗肿瘤活性的影响

目的:探讨不同溶剂对鸡血藤提取物中的总黄酮含量和抗肿瘤活性的影响。方法:分别用不同浓度的乙醇水溶液、丙酮水溶液作为提取溶剂,利用回流法、超声提取法制备鸡血藤提取物;采用四甲基噻唑蓝(MTT)法测定提取物对结肠癌HT-29细胞增殖的抑制活性;采用分光光度法测定提取物中总黄酮含量;采用HPLC法测定提取物中芒柄花素含量。结果:60%乙醇回流60 min所得提取物总黄酮含量最高并且对HT-29细胞增殖的抑制活性最强。结论:鸡血藤抗肿瘤活性提取物的制备以60%乙醇回流提取为佳。     

In vitro cytotoxic activity of some Korean medicinal plants on human cancer cell lines: enhancement in cytotoxicity by heat processing

The goal of this study was to examine the in vitro antiproliferative activity of crude methanol extracts of three traditional Korean medicinal plants: Achyranthes fauriei, Epimedium koreanum Nakai and Scutellaria baicalensis before and after heat processing. The extracts were screened for antitumoral potential by means of an MTT assay on four human cancer cell lines: lung cancer cells (Lu1), colon cancer cells (Col2), oral epidermoid carcinomas (KB) and hormone-dependent prostate cancer cells (LNCaP). None of the extracts showed significant activity against any of the cancer cell lines. However, after treatment with steam, the processed Achyranthes fauriei extract exhibited a slight, but enhanced cytotoxic activity against all four of the cancer cell lines. The processed Scutellaria baicalensis extract exerted a potent cytotoxic activity against the Lu1 cell line in a specific manner with an IC(50) value of 14.3 microg/mL. The Epimedium koreanum extract showed no cytotoxic effects against any of the cancer cell lines with or without heat processing. These results suggest that the heat processing of medicinal plants represents a possible route to the development of antitumor agents.     

Effect of Rhaphidophora korthalsii methanol extract on human peripheral blood mononuclear cell (PBMC) proliferation and cytolytic activity toward HepG2

The study of bioactivity of natural product is one of the major researches for drug discovery. The aim of this finding was to study the proliferation effect of Rhaphidophora korthalsii methanol extract on human PBMC and subsequently the cytotoxic effect of activated PBMC toward HepG2 human hepatocellular carcinoma. In this present study, MTT assay, cell cycle study and Annexin 5 binding assay were used to study the immunomodulatory and cytotoxic effects. In vitro cytotoxic screening of Rhaphidophora korthalsii methanol extract showed that the extract was non-toxic against hepatocellular carcinoma (HepG2). In contrast, the extract was able to stimulate the proliferation of human PBMC at 48 h and 72 h in MTT assay and cell cycle progress study. The application of immunomodulator in tumor research was studied by using MTT microcytotoxicity assay and flow cytometric Annexin V. Results indicated that pre-treated PBMC with Rhaphidophora korthalsii methanol extract induced the highest cytotoxicity (44.87 ± 6.06% for MTT microcytotoxicity assay and 51.51 ± 3.85% for Annexin V) toward HepG2. This finding demonstrates that Rhaphidophora korthalsii methanol extract are potent to stimulate the cytotoxic effect of immune cells toward HepG2.     

Study of the safety of oral Triphala aqueous extract on healthy volunteers

BackgroundTriphala extract is a well known medicinal herbal formula which is usually prescribed by Thai traditional doctors to adjust the physiological functions of the body. Previous studies have reported that Triphala has antioxidant, anti-inflammatory, antihypercholesterolemia and anticancer properties. Though this herbal recipe is commonly used in Thailand, its human safety, especially in the oral form, has not been studied. We therefore conducted a clinical trial (Phase I).ObjectiveThis study evaluated the safety of administering the aqueous extract of Triphala to healthy volunteers at 2500 mg/d.Design, setting, participants and interventionsAn open-label, single-arm trial was conducted at Chulabhorn International College of Medicine, Thammasat University, Pathum Thani, Thailand, between July 2017 and July 2018. The study enrolled 10 male and 10 female healthy volunteers; all were given Triphala (water extract; five capsules of 500 mg each) orally, once a day, at bedtime, for four consecutive weeks.Main outcome measuresSigns and symptoms, physical examinations, hematology and blood chemistry were assessed at the beginning of the trial and every week thereafter, for four consecutive weeks. After finishing the trial, on day 28, all volunteers were invited to a follow-up session on day 35 to evaluate the safety of the herbal recipe using the same measurements.ResultsAt the oral dose of 2500 mg/d, Triphala had no serious adverse effects in healthy volunteers. Moreover, it was found to have significantly improved the volunteers’ high-density lipoprotein cholesterol (HDL-C) levels on day 35 and also reduced their blood sugar levels on days 14 and 35.ConclusionsWe conclude that aqueous extract of Triphala is safe for healthy volunteers and that it elevates HDL-C levels and lowers blood sugar. Further clinical study should investigate its effects on HDL-C and blood sugar levels among the dyslipidemic and prediabetic groups.Trial registrationThis trial was registered in the Thai Clinical Trial Registry with the identifier TCTR20180423002.