Synergistic anticancer effect of the extracts from Polyalthia evecta caused apoptosis in human hepatoma(HepG2)cells

Objective:To evaluate the anticancer activity of the extract fraction of Polyalthia evecta(P. evecta)(Pierre) Finet & Gagnep and the synergistic anticancer effect of the extracts from P.evecta by using the ATR/FT-IR spectroscopy.Methods:The 50%ethanol-water crude leaf extract of P.evecta(EW-L) was prepared and was further fractionated to isolate various fractions.The anticancer activity was investigated from cytotoxicity against HepG2 using a neutral red assay and apoptosis induction by evaluation of nuclei morphological changes after DAPI staining. Synergistic anticancer effects of the extracts from P.evecta were performed using the ATR/FTIR spectroscopy.Results:The result showed that the EW-L showed higher cytotoxicity and apoptosis induction in HepG2 cells than its fractionated extracts.The hexane extract exhibited higher cytotoxicity and apoptosis induction than the water extracts,but less than the EW-L. The combined water and hexane extracts apparently increased cytotoxicity and apoptosis induction.The%apoptotic cells induced by the extract mixture were increased about 2-fold compared to the single hexane extract Conclusions:The polar extract fraction is necessary for the anticancer activity of the non-polar extract fraction.The ATR/FT-IR spectra illustrates the physical interaction among the constituents in the extract mixture and reveals the presence of polyphenolic constituents in the EW-L,which might play a role for the synergistic anticancer effect.

Sulforaphene in Raphanus sativus L. var. caudatus Alef increased in late-bolting stage as well as anticancer activity

Objectives: To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effect on HCT116 colon cancer cells.Methods: FTIR–ATR and GC–MS were used to characterize the isothiocyanates in the plant extracts followed by HPLC for quantification. Antiproliferation and apoptosis induction were determined by using MTT assay and flow cytometry, respectively.Results: The respective rank of anticancer activity of Raphanus sativus were as follows:vegetative(3 week) < older rosette(4 week) < early-bolting(5 week) < senescence(7 week) < late-bolting(6 week). The low to high concentration of sulforaphene and sulforaphane occurred in the same stage order.Conclusions: The reproductive parts(flower, pod, and dry seed) of Raphanus sativus have the greatest isothiocyanate concentration, evidenced by a sulforaphene concentration higher than the sulforaphane. This result should inform the selection of the most appropriate harvesting stage and plant part for use as a potential chemopreventive agent.

Biomolecular changes and DNA targeting effect of sesamol in human lung adenocarcinoma (SK-LU-1) cells by FTIR microscopy

Objective: To investigate biomolecular alteration of sesamol on human lung adenocarcinoma(SK-LU-1) cells compared with cisplatin using Fourier transform infrared microscopy(FTIR). Methods: Cytotoxicity of sesamol was investigated against SK-LU-1 cells by using neutral red. DNA fragmentation and the cell cycle analysis were determined by agarose gel electrophoresis and flow cytometry, respectively. The FTIR microscopy technique was applied to explore the changes in cellular biochemical compositions in cells treated with sesamol that the biochemical and biological assays cannot cover. The alkylating property was determined by 4-(4-nitrobenzyl)pyridine assay. Results: Sesamol and cisplatin exerted an antiproliferative effect at 48 h with respective IC50 values of 2.7 and 0.07 m M. Both induced apoptosis by causing DNA damage and accumulation of cell populations at sub-G1. FTIR microscopy and Principle Component Analysis clearly discriminated the sesamol-and cisplatin-treated cells from the untreated cells or control. A significant increase of total lipid content was found in cisplatin-treated cells. Conformational changes in the proteins secondary structure from the β-helix to the β-sheet were found in both sesamol-and cisplatin-treated cells, as well as significant reductions in relative DNA content of both compared to the control were observed, suggesting DNA damage. A shift in the peak position of DNA region provides insight on the DNA interactions. Conclusions: The non-alkylating effect of sesamol based on the nitrobenzyl pyridine assay delineates the non-covalent binding mode of sesamol on DNA. Hydrogen bonding is the binding mode of sesamol on DNA, while for cisplatin it was covalent and hydrogen bonding.

Effects of Heliotropium indicum L.on Uterine Involution and Its Underlying Mechanisms:An in vivo and in vitro Study

Objective:To investigate the effect of Heliotropium indicum L.(H.indicum L.)on uterine involution and its underlying mechanisms in both in vivo and in vitro study.Methods:For in vivo studies,postpartum rats were randomly divided into 2 groups(n=24 for each):control group and treated group which were orally and daily administered with ethanolic extract of H.indicum L.(250 mg/kg body weight)until day 5 of postpartum.Uteri were collected for analysis of weight,cross-sectional area,collagen cross-sectional area,and collagen content on postpartum day 1,3,and 5(n=8 for each)from both groups.Blood samples were collected for hepatotoxicity and 17?-estradiol(E_(2))measurement.For in vitro studies,the extract effects on uterine contraction at half maximum effective concentration of 2.50 mg/mL were studied in organ bath system for at least 20 min.Results:Uterine parameters were significantly decreased after treated with extract of H.indicum L.(P<0.05).H.indicum L.extract significantly accelerated the reduction of those parameters and significantly decreased E_(2)(P<0.05).The extract facilitated uterine involution with no hepatotoxicity.H.indicum L.extract significantly stimulated uterine contraction(P<0.05)and synergized with oxytocin,prostaglandin and its precursor,linoleic acid.By investigating the different sequencing of the extract with the additional stimulants(added before or after),the two showed antagonistic effects,but still showed potentiated force when compared with control(without the stimulants).Conclusions:The underlying mechanisms by which H.indicum L.facilitated uterine involution might be due to reducing E_(2)which induces collagenase activity,leading to decreases in uterine weight and size and stimulating uterine contraction.Our study provides new findings for future drug development for facilitating uterine involution with H.indicum L.