Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-α, and angiogenesis by blocking endothelial functions

OBJECTIVE: The present study aimed to investigate the mechanisms underlying the anti-inflammatory and anti-angiogenic effects of ethyl-p-methoxycinnamate isolated from Kaempferia galanga. METHODS: The anti-inflammatory effects of ethyl-p-methoxycinnamate were assessed using the cotton pellet granuloma assay in rats, whereby the levels of interleukin-1 and tumor necrosis factor-α were measured in the animals' blood. In addition, the levels of interleukin, tumor necrosis factor, and nitric oxide were measured in vitro using the human macrophage cell line (U937). The analgesic effects of ethyl-p-methoxycinnamate were assessed by the tail flick assay in rats. The anti-angiogenic effects were evaluated first by the rat aortic ring assay and, subsequently, by assessing the inhibitory effects of ethyl-p-methoxycinnamate on vascular endothelial growth factor, proliferation, migration, and tube formation in human umbilical vein endothelial cells. RESULTS: Ethyl-p-methoxycinnamate strongly inhibited granuloma tissue formation in rats. It prolonged the tail flick time in rats by more than two-fold compared with the control animals. The inhibition of interleukin and tumor necrosis factor by ethyl-p-methoxycinnamate was significant in both in vivo and in vitro models; however, only a moderate inhibition of nitric oxide was observed in macrophages. Furthermore, ethyl-p-methoxycinnamate considerably inhibited microvessel sprouting from the rat aorta. These mechanistic studies showed that ethyl-p-methoxycinnamate strongly inhibited the differentiation and migration of endothelial cells, which was further confirmed by the reduced level of vascular endothelial growth factor. CONCLUSION: Ethyl-p-methoxycinnamate exhibits significant anti-inflammatory potential by inhibiting pro-inflammatory cytokines and angiogenesis, thus inhibiting the main functions of endothelial cells. Thus, ethyl-p-methoxycinnamate could be a promising therapeutic agent for the treatment of inflammatory and angiogenesis-related diseases

In vitro antimetastatic activity of Agarwood (Aquilaria crassna) essential oils against pancreatic cancer cells

Background: Pancreatic cancer is one of the most lethal malignant tumors which remains a rampant killer across the globe. Lack of early diagnosis and toxic drugs have failed to improve the survival rate of pancreatic cancer patients, thus new agents that are safe, available and effective are urgently needed. Objective: The study aimed to investigate the efficacy of Agarwood essential oils in the inhibition of metastasis and induction of apoptosis in the pancreatic cell line (MIA PaCa-2). Methods: Essential oils of Aquilaria crassna were obtained by hydrodistillation. Chemical characterization was analyzed using FTIR and GCMS. The effects of essential oils against three steps of metastases have been investigated, including cell proliferation, migration and clonogenicity. Hoechst and rhodamine assays confirmed the mechanism of pancreatic cancer cell death. Results: The results showed that essential oils exhibited potent cytotoxic activity against MIA PaCa-2 cells with an IC50 (11 ± 2.18 μg/ml). Cell migration was effectively inhibited at (10 μg/ml). Moreover, at a sub-toxic dose (5 μg/mL), essential oils obstructed the colony formation properties of MIA PaCa-2 significantly. The mechanism of cell death was determined due to the induction of nuclear condensation and disruption of mitochondrial membrane potential in the cells. Interestingly, several active components were existed in the chemical profile of the essential oils extract such as β-Caryophyllene, 1-Phenanthrenecarboxylic acid, azulene, naphthalene and Cyclodecene. Conclusion: The present study elucidated for the first time the anti-pancreatic cancer properties of A. crassna essential oils, It can be concluded that the anticancer effects of the extract could be due to the synergistic effect of the biologically active phytoconstituents present in the essential oils

β-Caryophyllene Induces Apoptosis and Inhibits Angiogenesis in Colorectal Cancer Models

Beta-Caryophyllene (BCP), a naturally occurring sesquiterpene abundantly found in cloves, hops, and cannabis, is the active candidate of a relatively new group of vascular-inhibiting compounds that aim to block existing tumor blood vessels. Previously, we have reported the anti-cancer properties of BCP by utilizing a series of in-vitro anti-tumor-related assays using human colorectal carcinoma cells. The present study aimed to investigate the effects of BCP on in-vitro, ex-vivo, and in-vivo models of anti-angiogenic assays and evaluate its anti-cancer activity in xenograft tumor (both ectopic and orthotopic) mice models of human colorectal cancer. Computational structural analysis and an apoptosis antibody array were also performed to understand the molecular players underlying this effect. BCP exhibited strong anti-angiogenic activity by blocking the migration of endothelial cells, tube-like network formation, suppression of vascular endothelial growth factor (VEGF) secretion from human umbilical vein endothelial cells and sprouting of rat aorta microvessels. BCP has a probable binding at Site#0 on the surface of VEGFR2. Moreover, BCP significantly deformed the vascularization architecture compared to the negative control in a chick embryo chorioallantoic membrane assay. BCP showed a remarkable reduction in tumor size and fluorescence molecular tomography signal intensity in all the mice treated with BCP, in a dose-dependent relationship, in ectopic and orthotopic tumor xenograft models, respectively. The histological analysis of the tumor from BCP-treated mice revealed a clear reduction of the density of vascularization. In addition, BCP induced apoptosis through downregulation of HSP60, HTRA, survivin, and XIAP, along with the upregulation of p21 expressions. These results suggest that BCP acts at multiple stages of angiogenesis and could be used as a promising therapeutic candidate to halt the growth of colorectal tumor cells

Crystal Structure Elucidation and Anticancer Studies of (-)-Pseudosemiglabrin: A Flavanone Isolated from the Aerial Parts of <i>Tephrosia apollinea</i>

<div><p><i>Tephrosia apollinea</i> is a perennial shrublet widely distributed in Africa and is known to have medicinal properties. The current study describes the bio-assay (cytotoxicity) guided isolation of (-)-pseudosemiglabrin from the aerial parts of <i>T. apollinea</i>. The structural and stereochemical features have been described using spectral and x-ray crystallographic techniques. The cytotoxicity of isolated compound was evaluated against nine cancer cell lines. In addition, human fibroblast was used as a model cell line for normal cells. The results showed that (-)-pseudosemiglabrin exhibited dose-dependent antiproliferative effect on most of the tested cancer cell lines. Selectively, the compound showed significant inhibitory effect on the proliferation of leukemia, prostate and breast cancer cell lines. Further studies revealed that, the compound exhibited proapoptotic phenomenon of cytotoxicity. Interestingly, the compound did not display toxicity against the normal human fibroblast. It can be concluded that (-)-pseudosemiglabrin is worthy for further investigation as a potential chemotherapeutic agent.</p></div

IC₅₀ Values of (-)-Pseudosemiglabrin (SSG) on Various Human Cancer Cell Lines^a.

a<p>The median inhibitory concentrations (IC₅₀) were determined by nonlinear regression analysis of log-concentration-response curves of 3 different tests (n = 3).</p

FT-IR spectral features of (-)-pseudosemiglabrin.

<p>The figure highlights alkoxy (1), C = C aromatic (2), carbonyl (3), and alkyl group (4) stretches of SSG.</p

Dose-dependent anti-proliferative effect of SSG on, HCT 116, MCF-7, PC3, HL-60, K562, 4T1, HT-29, U937 and CCD cell lines was assessed by MTT-assay.

<p>(Values are represented as mean ± SD, n = 3).</p

ORTEP (Oak Ridge Thermal Ellipsoid plot) picture of (-)-pseudosemiglabrin with the displacement ellipsoid drawn at 50% probability.

<p>The figure presents two crystallographically different molecules (A and B) of title compound.</p

A) The photomicrographs depict the images of PC3 cells with Hoechst 33258 stain taken at 6 and 12 hours after treatment.

<p>The cells treated with SSG revealed clear signs of proapoptosis. The cells treated with 0.1% DMSO (Vehicle) showed prompt and evenly distributed nucleus with fully extended pseudopodial like projections of cell membrane. Whereas, the cells treated with SSG (10 µM) displayed blebbing of cellular membrane and the typical apoptotic changes in the chromatin structure. The arrows indicate the clear signs of nuclear condensation including the half moon (crescent) shaped apoptotic nuclei. The arrowheads indicate the chromatin dissolution, breakdown and fragmentation. The standard reference, betulinic acid also showed the similar induction of apoptosis in the cells. B) Graphical representation of percentage of apoptotic indices. The apoptotic index for each test group was expressed as a percentage of the ratio of apoptotic cells number to the total cell number in 10 different fields. Values are presented as mean ± SD (<i>n</i> = 10), * represents p<0.05 and ** represents p<0.01.</p

Stereochemical structures of (-)-semiglabrin and SSG.

<p>Stereochemical structures of (-)-semiglabrin and SSG.</p