Echinacea is a North American native medicinal herb used traditionally for wounds, burns, snake or insect bites, colds, infections, and inflammation by indigenous Americans. We investigated the effects of the root and stem of fresh Korean-grown Echinacea angustifolia methanol extracts and fractionation extracts on the cytotoxicity against cancer cells (HL60, 3LL). The extracts were prepared by step-wise fractionation of methanol extracts of Echinacea angustifolia using hexane, chloroform, ethyl acetate, buthanol, and water. From the tests, root and stem parts of Echinacea showed the cytotoxic effect on cancer cells. The cytotoxie rate of the ethyl acetate fraction of the root parts showed 79% against HL60 cells at low concentration (0.125 mg/mL), and hexane fraction of the root and stem parts gradually increased as the concentration of samples increased, and the root parts showed 82% at 1.0 mg/mL concentration against HL60 cells, chloroform fraction of the root part showed 78.4% against HL60 cells and 68.4% on 3LL cells at 1.0 mg/mL concentration, water and butanol fraction of these results, it is considered that ethyl acetate fraction of the root and stem parts showed 60.1% to 77.1% against HL60 cells, after testing by MTT assay system. From these results, it is considered that ethyl acetate fraction of the Echinacea angustifolia root parts has stronger anticancer effects than any other fractions in vitro.
Recently, we reported that an ethanol extract of Iris nertschinskia induces p53-dependent apoptosis in the MCF7 human breast cancer cell line. However, the detailed mechanisms were not fully explored. Here, we demonstrate another aspect of the activity of I. nertschinskia in breast cancer cells. We compared the response to an ethanol extract of I. nertschinskia in two different human breast cancer cell lines, Hs578Tand MDA-MB231, respectively with relatively low and high AKT1/2 activity by trypan blue exclusion assay and FACS analysis. Knockdown of endogenous AKT1 or AKT2 in breast cancer cells by RNA interference determined the sensitivity to I. nertschinskia ethanol extract compared to control cells. The I. nertschinskia ethanol extract induced cell death in a manner that depended on the level of phosphorylated AKT1/2 protein and was associated with a significant increase in the sub-G1 cell population, indicative of apoptosis. Our results indicate that an ethanol extract of I. nertschinskia differentially induces cell death in breast cancer cells depending on their level of phosphorylated AKT1/2.
Jeon, Ye Won;Cho, Sun Hang;Han, Hee Dong;Shin, Byung Cheol
Journal of the Korean Chemical Society
/
v.58
no.6
/
pp.575-579
/
2014
Development of liposomes has been actively studied for effective delivery of drug at tumor site. However, despite their preferential accumulation at tumor site, the therapeutic efficacy of such liposomal drug has been limited because of low drug release. Therefore, we developed a temperature-sensitive liposome (TSL), which can be made to maximize release of drug by external stimulation such as ultrasound. Doxorubicin (DOX) as a model drug was encapsulated into TSL by a pH gradient method. The particle size of the TSL was $142.0{\pm}6.24nm$. Surface charge was $-10.55{\pm}1.12mV$. Release of drug from TSLs was up to 80% within 15 min at over $42^{\circ}C$ measured by fluorescence intensity. Cytotoxicity of released DOX from TSLs with ultrasound was highly increased compared to TSLs without ultrasound. Taken together, we demonstrate that temperature sensitive drug release from TSLs with ultrasound, which may be useful for cancer therapy to increase drug concentration at tumor site by external stimulation.
The objectives of this study were to examine the lung injury and inflammation caused by manual metal arc stainless steel(MMA-SS) welding fume inhalation and to evaluate the recovery process. Sprague-Dawley rats were exposed to MMA-SS welding fumes for 2 h per day in a whole-body exposure chamber, with a total suspended particulate(TSP) concentration of $51.4{\pm}2.8mg/m^3$(low dose) or $84.6{\pm}2.9mg/m^3$(high dose) for 30 days. The animals were sacrificed after 30 days of exposure as well as after a 30-day recovery period. To assess the inflammatory or injury responses, cellular and biochemical parameters as well as cytokines were assayed in the bronchoalveolar lavage fluid(BALF). MMA-SS welding fume exposure led to a significant elevation in the number of alveolar macrophages(AM) and polymorphonuclear cells(PMN). Additionary, the values of $\beta$-n-acetyl glucosaminidase($\beta$-NAG) and lactate dehydrogenase(LDH) in the BALF were increased in the exposed group when compared to controls. After 30 days of recovery from exposure, a significant reduction in inflammatory parameters of BALF was observed between the exposed and recovered groups. Slight, but significant elevations were noted in the number of AM and PMN in the recovered groups, and AM that had been ingested fume particles still remain in the lungs. In conclusion, these results indicated that welding fumes induced inflammatory responses and cytotoxicity in the lungs of exposed rats. Fume particles were not fully cleared from lungs even after a 30-day recovery period.
This study was carried out to investigate the influence of stem materials such as black stem on the quality of expended stem and cigarettes. Normal and black stem were separated by tobacco scan and then, those stems were expanded after treating with their respective stem casings. Total sugar, ether extract, ash contents and pH were slightly low in black stem compared with normal stem. However, the number of bacteria and fungi ratio were remarkably higher in black stem than that of normal stem. As compared with normal stems, ratio of rushed stem in rolled process was approximately 2 times higher in black stem with the consequency that the filling capacity of black stem was decreased. The ratio of large particles (> 3.35 mm) of expanded black stem showed decreasing tendency and small particles rate (1.40 mm <) was increased compared with normal stem. When expanded stems were prepared using stem containing 5 levels (0, 10, 20, 30 and 100 %) of black stem, the filling capacity was decreased and static burning rate was significantly decreased with increasing expanded black stem rate. However, the weight and hardness of cigarettes were slightly increased with increasing expanded black stem rate. The contents of phenol compounds, aromatic amines and carbonyl compounds in the cigarette mainstream smoke from the cigarette which was manufactured with various ratio of expended black stem, were gradually increased with increasing expanded black stem rates. Also, the cytotoxicity and the mutagenicity of the TPM were significantly increased with increasing expanded black stem rate. The sensory test result showed that cigarettes blended with 10 and 30 % level of black stem rate was exhibited significantly high sensory attributions such as off-taste, impact, hotness, bitterness and irritation as compared with cigarette blended with normal stem, while smoke fullness and cleanness were slightly decreased with increasing expanded black stem rates. The number of brown spots on cigarettes paper was 2 to 3 times high in cigarettes containing black stem than that of cigarette made from normal stem and were high with increasing black stem rate. The overall assessment in this study suggest, that black stem should not be used because of bad quality of expanded stem and high toxicological activity of cigarette mainstream smoke.
Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.
The immune activation activities of ethanol, ethanol : water(1 : 1,v/v) and water extracts from Dendropanax morbifera Lev. were compared. The crude extracts showed relatively low cytotoxicity as less than 26% by using human normal liver cell(WRL-68). For example, the ethanol extract inhibited. in MCF7 and Hep3B cells in adding 1.0 mg/mL ethanol extracts. The growth of human immune T and B cells was enhanced up to $1.22{\sim}1.27$ times by adding the crude extracts. These are a trend of increasing secretion of $cytokines(IL-6,\;TNF-{\alpha})$ from human B/T cell for 6 day cultivation. It was found that the immune activation activities of the crude extracts seemed to be higher than those obtained by using other solvents.
The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelial-mesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and anti-metastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.
Anti-angiogenesis and anti-cell adhesion effects were investigated with different dose of Camellia japonica leaf (CJL) extract for applying anti-cancer, anti-metastasis and anti-obesity. Cytotoxicity on HUVECs was very low at 200 ug/mL of CJL-extract. Anti-angiogenic ratio at increasing dose of 1.5 ug/mL, 3.0 ug/mL, 15 ug/mL and 30 ug/mL was showed 30.7%, 38.5%, 53.8%, and 70.0%, respectively. Also, anti-cell adhesion effect at concentration of $50{\mu}g{/well},\;100{\mu}g{/well}\;and\;200{\mu}g{/well}$ was expressed on E-selectin by 46.7%, 66.7%, and 86.76%, on VCAM-1, 23.0%, 61.5%, and 84.6%, and on ICAM-1, 11%, 55.5%, and 88.8%, respectively. For inquiring anti-angiogenesis mechanism, when western blot was performed with different dose of CJL extract, signal molecules of VEGFR-2, $\beta$-catenin and PI3-K were suppressed. As the signal transduction from VEGFR-2, $\beta$-catenin and PI3-K to NF-${\kappa}$B was interupted, angiogenesis could not be occurred causing not activated NF-kB. C. japonica leaf (CJL) is a useful herb for developing therapeutics of angiogenesis related diseases such as cancer, metastasis, rheumathioid arthritis and obesity.
Objectives : In order to investigate the anti-obesity effects of Wolbi-tang(here in after referred to WBT) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high-fat diet. Methods : C57BL/6 mice were divided into 5 groups(normal, only high-fat diet, high-fat diet with Reductil, high-fat diet with WBT 400, 200 mg/kg extract) and fed for 5 weeks. And observed body weight change, total cholesterol, low density lipoprotein cholesterol(LDL-cholesterol), high density lipoprotein cholesterol (HDL-cholesterol), triglyceride, glucose, leptin change, alanine transaminase(ALT), aspartate transaminase(AST), serum creatinine, the expression of ${\beta}3$-adrenergic receptor(${\beta}3AR$), leptin, uncoupling protein(UCP2) gene in 3T3-L1 adipocyte, 3T3-L1 adipocyte proliferation, histological analysis of adipose tissue and liver tissue. Results : 1. Refer to cell cytotoxicity, viability of human fibroblast cells(hFCs) showed not significant changes. 2. The amount of ALT, AST was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. The amount of creatinine showed not significant changes. 3. Body weight was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 4. The amount of total cholesterol and triglyceride was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. LDL-cholesterol was decreased and HDL-cholesterol was increased significantly in WBT 400 mg/kg groups. 5. The amount of glucose was decreased significantly in WBT 400 mg/kg groups. 6. The amount of serum leptin was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 7. The revelation of ${\beta}3AR$ in 3T3-L1 adipocyte was increased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of leptin was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of UCP2 was decreased significantly in WBT $100{\mu}g/ml$ group. 8. 3T3-L1 adipocyte proliferation was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The size of adipocyte was decreased relative to the control group in WBT 400 mg/kg group. 9. The adipose vacuoles in liver tissue was decreased relative to the control group. Conclusions : These results suggested that WBT has inhibitory effects of obesity. WBT might be applicated on treatment of obesity and metabolic syndrome. Further studies analysing its effects were needed.
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