Monday, December 10, 2012

CAPE(caffeic acid phenethyl ester)'s affect on growth

CAPE (caffeic acid phenethyl ester) is a SERM and is a component of bee propolis.  It's unclear what effect CAPE has on growth.

Caffeic acid 3,4-dihydroxy-phenethyl ester induces cancer cell senescence by suppressing twist expression.

"Unlike caffeic acid phenethyl ester, caffeic acid 3,4-dihydroxy-phenethyl ester (CADPE), which is isolated from the medicinal plants Sarcandra glabra and Teucrium pilosum, inhibits human cancer cell growth and colony formation by inducing cancer cell senescence, not apoptosis. CADPE induces cell senescence and morphology changes by increasing cellular size and cytoplasmic granularity, enhancing senescence-associated β-galactosidase activity and differentiated embryo-chondrocyte expressed gene 1 expression, and blocking cell-cycle arrest in the G(1) phase {maybe CAPDE can increase chondrocyte hypertrophy?}. CADPE significantly suppressed the expression of Twist1{Twist1 does inhibit the early stages chondrogenesis} and led to the up-regulation of rat sarcoma, p53, p21(WAF1/CIP1), and p16(INK4a) proteins in a dose-dependent manner, resulting in the hypophosphorylation of retinoblastoma protein. Overexpression of Twist1 prevented CADPE-induced cell senescence in tumor cells. CADPE [targets] the Twist1-dependent senescence signaling pathway."

Overexpression of Twist1 tends to cause overgrowth by inhibiting cellular senescence.  Since CAPDE inhibits Twist1, CAPDE may not be good for height growth.

"senescent cells develop a distinct and recognizable flattened and enlarged morphology with a prominent nucleus and increased cytoplasmic granularity. "

"STAT3 can up-regulate Twist1 expression"

"CADPE has little effect on Twist1 low expression normal cells"<-Growth plates do express Twist1 so that may be sufficient to be affected by CADPE.

Effect of propolis on human cartilage and chondrocytes.

"Propolis [is] a natural product derived from plant resins collected by the honeybees. The extract that contains amino acids, phenolic acids, phenolic acid esters, flavonoids, cinnamic acid, terpenes and caffeic acid, possesses several biological activities such as anti-inflammatory, immunostimulatory, anti-viral and anti-bacterial. We assay the effects of propolis extract on the production of key molecules released during chronic inflammatory events as nitric oxide (NO) and glycosaminoglycans (GAGs) in cultures of human cartilaginous tissues and chondrocytes, stimulated with interleukin-1beta (IL-1beta). This natural compound and its active principle, caffeic acid phenethyl ester (CAPE), were able to contrast the harmful effects of IL-1beta."

CAPE alone did not affect NO synthesis but did blunt the NO levels induced by IL-1B.

"peroxynitrite (ONOO-), generated by the reaction of NO with superoxide anion (O2−), stimulates COX-2 activity and inhibits the proteoglycan synthesis in rat chondrocytes stimulated with IL-1β"

So CAPE is an anti-inflammatory agent.  Inflammation is more beneficial with closed growth plates than open growth plates so CAPE may help protect against excess NO production in open growth plates.

Caffeic acid phenethyl ester, a component of beehive propolis, is a novel selective estrogen receptor modulator.

"Caffeic acid phenethyl ester (CAPE) is an active ingredient of beehive propolis with a structure similar to phenolic acid. CAPE showed selective binding affinity to human estrogen receptor beta (hERbeta) rather than hERalpha. CAPE also reduced ERalpha expression in MCF-7 and MDA 231 cells. In the yeast estrogen receptor transcription assay, CAPE produced the transcriptional activity of estrogen-responsive element with EC(50) values of 3.72 x 10(-6) M. CAPE did not increase the growth of MCF-7 estrogen receptor-positive breast cancer cells in doses ranging from 10(-7) to 10(-5) M. In order to understand how CAPE acts in animals, CAPE was tested by a uterotrophic bioassay. Treatment with CAPE (100, 500 mg/kg) did not increase the uterine weight relative to 3 microg/kg 17beta-estradiol treatment. CAPE is a selective agonist to hERbeta."

Royal jelly on the other hand binds to ERalpha.  Is the binding with ERBeta competitive?  Does CAPE inhibit growth decreasing agents from binding to ERBeta?

"Anthropometric measurements and gene expression were measured in 139 female offspring. The two length and the height measurements were correlated, as were BMI and weight. CYP19 expression was correlated with the other gene expressions and both estrogen receptor expressions were associated. For every 1 unit of ΔCt (18SrRNA ‐ CYP19) or ΔCt (RNA PolII ‐ CYP19), BMI was increased by 0.9 and 0.87 kg/m2, respectively, and weight by 2.35 kg and 2.1 kg, respectively. For every 1 unit of ΔCt (18SrRNA ‐ CYP17), leg length was increased by 0.84 cm. CYP17[17α hydroxylase] gene expression may influence growth during childhood and adolescence."

"CYP17 deficiencies in patients due to genetic mutations [resulted in] individuals [with] tall stature and retardation of bone age"

"one particular polymorphism on the promoter region of CYP17 is known to increase CYP17 activity [which also results in increased height]"

"Average height and trunk length were not significantly related to the expression of any of the four genes[ERalpha, ERbeta, Cyp17, Cyp19]."

Although the trend was for ERbeta to be more beneficial to height than ERalpha.

They also used peripheral blood cells to detect for gene expression rather than epiphyseal growth plates.

Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-kappa B. states that CAPE is also involved inhibiting the translocation of NFkB to the nucleus.

LSJL upregulates ESR2(ERBeta).

"two SNPs of IGF1, rs5742694 and rs2033178, were significantly associated with human heights. For ESR2, significant associations were only detected in women (rs1256061; rs17766755; rs1256044). No association was detected between CYP17 and human height."

So basically CAPE enhances ERbeta and the role of ERbeta in male height is unclear.

Inhibition of Ca(2+) release-activated Ca(2+) channel (CRAC) by curcumin and caffeic acid phenethyl ester (CAPE) via electrophilic addition to a cysteine residue of Orai1.:  CAPE inhibits calcium channels in kidney cells.

Suppression of Toll-like receptor 4 activation by caffeic acid phenethyl ester is mediated by interference of LPS binding to MD2.: "CAPE suppressed production of inflammatory mediators and activation of NFκB and IRF3 in macrophages stimulated with LPS. CAPE interrupted LPS binding to MD2 through formation of adduct specifically with Cys133 located in hydrophobic pocket of MD2."  "CAPE [prevents] TLR4 activation by interfering with interaction between ligand(LPS) and receptor complex(TLR4/MD2)."

Comparative Analysis of the Protective Effects of Caffeic Acid Phenethyl Ester (CAPE) on Pulmonary Contusion Lung Oxidative Stress and Serum Copper and Zinc Levels in Experimental Rat Model.:  CAPE lowered copper and zinc levels in lung cells.

CAPE suppresses VEGFR-2 activation, and tumor neovascularization and growth.: "CAPE suppressed VEGF-induced proliferation, tube formation, migration, the formation of actin stress fibers and loss of VE-cadherin at cell-cell contacts in endothelial cells, indicating the inhibition of VEGF-mediated VEGF receptor-2 (VEGFR-2) and its downstream signal activation in vitro. CAPE blocked VEGF-stimulated neovascularization in the Matrigel plugs assay, and reduced vascular permeability in mouse skin capillaries in vivo. CAPE inhibited the growth and neovascularization of primary tumor cells in C57BL/6 and BALB/c mice inoculated with Lewis lung carcinoma, colon carcinoma, and melanoma cells."<-this is for normal epithelial cells.  Will this inhibition of VEGFR2 translate to chondrocytes.

Grape seed proanthocyanidins inhibit migration potential of pancreatic cancer cells by promoting mesenchymal-to-epithelial transition and targeting NF-κB.

"Here we explore the effect of grape seed proanthocyanidins (GSPs) on pancreatic cancer cell migration and the molecular mechanisms underlying these effects. Treatment of human pancreatic cancer cell lines Miapaca-2, PANC-1 and AsPC-1 with GSPs resulted in inhibition of cell migration (19-82%), which was associated with decreased phosphorylation of ERK1/2 and inactivation of NF-κB. Treatment of cells with UO126, an inhibitor of MEK, and caffeic acid phenethyl ester, an inhibitor of NF-κB, also inhibited the migration of cells (40-80%). Inhibition of cell migration by GSPs was associated with reversal of the epithelial-to-mesenchymal transition. This was associated with upregulation of E-cadherin and desmoglein-2 and down-regulation of fibronectin, N-cadherin and vimentin."

So CAPE helps prevent dedifferentiation.  Some studies have also showed that CAPE has an anti-proliferative effect it may do that by encouraging differentiation.

Not only melatonin but also caffeic acid phenethyl ester protects kidneys against aging-related oxidative damage in Sprague Dawley rats.

"Microscopic features and antioxidant status of kidneys of young, old, and caffeic acid phenethyl ester (CAPE) and melatonin administered old Sprague Dawley rats were evaluated. Aging-related tubular and glomerular changes were evident. The most prominent tubular alterations were massive vacuole formation, mitochondrial degeneration, and lysosome accumulation. Mean tissue malondialdehyde (MDA) level was increased, mean tissue superoxide dismutase (SOD), catalase (CAT) (p < .001), and glutathione peroxidase (GPx) activities (p < .05), and total glutathione (GSH) level were decreased in old animals. Melatonin significantly reduced tissue MDA levels (p < .005), but increased tissue SOD (p < .001), CAT, and GPx activities (p < .05), and GSH levels (p < .005) in old animals. CAPE also significantly reduced tissue MDA levels (p < .005), but increased tissue SOD (p < .05), CAT (p < .005), GPx activities, and GSH levels (p < .001) in old rats. Mean tissue MDA levels of melatonin and CAPE-administered rats were even lower than those of young rats (p < .05). In conclusion, tubular and glomerular structures and tissue antioxidant enzyme activities were very well preserved in CAPE and melatonin-administered rats."

Caffeic acid phenethyl ester suppresses the proliferation of human prostate cancer cells through inhibition of p70S6K and Akt signaling networks.: CAPE suppresses Akt signaling in cancer cells.

Caffeic acid phenethyl ester-mediated Nrf2 activation and IkappaB kinase inhibition are involved in NFkappaB inhibitory effect: structural analysis for NFkappaB inhibition.

"CAPE attenuated expression of NFkappaB dependent luciferase stimulated with TNF-alpha or LPS and suppressed LPS-mediated induction of iNOS, a target gene product of NFkappaB. In HCT116 cells, CAPE interfered with TNF-alpha dependent IkappaBalpha degradation and subsequent nuclear accumulation of p65, which occurred by direct inhibition of inhibitory protein kappaB kinase (IKK). CAPE increased the expression of Nrf2-dependent luciferase and heme oxygenase-1, a target gene of Nrf2, and elevated the nuclear level of Nrf2 protein, indicating that CAPE activated the Nrf2 pathway. In HCT116 cells with stable expression of Nrf2 shRNA, CAPE elicited a reduced inhibitory effect on TNF-alpha-activated NFsmall ka, CyrillicB compared to scramble RNA expressing control cells. On the other hand, the NFkappaB inhibitory effect of CAPE was diminished by removal or modification of the Michael reaction acceptor, catechol or phenethyl moiety in CAPE. CAPE inhibits TNF-alpha-dependent NFkappaB activation via direct inhibition of IKK as well as activation of Nrf2 pathway, in which the functional groups in CAPE may be involved."

Caffeic acid phenethyl ester accumulates beta-catenin through GSK-3beta and participates in proliferation through mTOR in C2C12 cells.:  In cancer cells, CAPE increased GSK3B phosphorylation and decreased Beta-Catenin phosphorylation.  CAPE stimulated the ERK pathway through the mTor pathway.

Caffeic acid phenethyl ester inhibits osteoclastogenesis by suppressing NF kappaB and downregulating NFATc1 and c-Fos.

"CAPE potently suppressed osteoclastogenesis in cultures of bone marrow-derived precursor cells with the osteoclast differentiation factor, receptor activator of nuclear factor kappaB ligand (RANKL). While the RANKL-stimulated activation of the ERK, JNK, and p38 MAPK signaling pathways was not affected, the DNA binding and transcription activity of NF kappaB were reduced by CAPE treatment. In addition, CAPE blocked the induction of NFATc1 and c-Fos following RANKL stimulation. Forced expression of c-Fos could reverse the inhibitory effect of CAPE on osteoclastogenesis. Finally, CAPE significantly inhibited the RANKL-induced osteoclast formation in mouse calvariae in vivo."

"c-Fos induces NFATc1 expression by directly binding to the promoter region of NFATc1"

6-8 week old female mice were used.

N-acetylcysteine stimulates osteoblastic differentiation of mouse calvarial cells.:  Unlike NAC, CAPE did not stimulate Osteoblast differentiation.

Antiplatelet activity of caffeic acid phenethyl ester is mediated through a cyclic GMP-dependent pathway in human platelets.: "Rapid phosphorylation of a platelet protein of Mw. 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12, 13-dibutyrate (150 nM). This phosphorylation was markedly inhibited by CAPE (15 and 25 microM). The present study reports a novel and potent antiplatelet agent, CAPE, which involved in the following inhibitory pathways: CAPE increases cyclic GMP/VASP Ser157 phosphorylation, and subsequently inhibits protein kinase C activity, resulting in inhibition of P47 phosphorylation, which ultimately inhibits platelet aggregation. "

Stimulatory actions of caffeic acid phenethyl ester, a known inhibitor of NF-kappaB activation, on Ca2+-activated K+ current in pituitary GH3 cells.

"Caffeic acid phenethyl ester (CAPE), a phenolic antioxidant derived from the propolis of honeybee hives, is known to be an inhibitor of activation of nuclear transcript factor NF-kappaB. Its effects on ion currents have been investigated in pituitary GH(3) cells. This compound increased Ca(2+)-activated K(+) current (I(K(Ca))) in a concentration-dependent manner with an EC(50) value of 14 +/- 2 microm. However, the magnitude of CAPE-induced stimulation of I(K(Ca)) was attenuated in GH(3) cells preincubated with 2,2'-azo-bis-(2-amidinopropane) hydrochloride (100 microm) or t-butyl hydroperoxide (1 mm). CAPE (50 microm) slightly suppressed voltage-dependent L-type Ca(2+) current. In inside-out configuration, CAPE (20 microm) applied to the intracellular face of the detached patch enhanced the activity of large conductance Ca(2+)-activated K(+) (BK(Ca)) channels with no modification in single-channel conductance. After BK(Ca) channel activity was increased by CAPE (20 microm), subsequent application of nordihydroguaiaretic acid (20 microm) did not further increase the channel activity. CAPE-stimulated channel activity was dependent on membrane potential. CAPE could also increase Ca(2+) sensitivity of BK(Ca) channels in these cells. Its increase in the open probability could primarily involve a decrease in the mean closed time. In current-clamp conditions, CAPE hyperpolarized the membrane potential and reduced the firing of action potentials. The stimulatory effects on these channels may partly contribute to the underlying mechanisms through which this compound influences the functional activities of neurons or neuroendocrine cells. Caution has to be used in attributing its response in the activation of NF-kappaB."

Caffeine dampens Ca2+ signaling so this is surprising.

The effect of selective oestrogen receptor antagonists in an in vitro model of growth plate chondrogenesis.

"We utilized an in vitro model of chondrogenesis, the RCJ3.1C5.18 cell line, to explore the effect of oestrogen on this process. We demonstrated the presence of oestrogen receptors (ER) α and β in these cells, with increased abundance of both receptor sub-types evident as the cells differentiated. ERα localized to the nucleus, suggesting it was signalling by genomic pathways, while ERβ was seen predominantly in the cytoplasm, suggesting it may be utilizing non-genomic signalling. While exogenous oestrogen had no effect on proliferation or differentiation, we found some evidence for the endogenous production of oestrogen (intracrinology), as suggested by the expression of aromatase in these cells. Selective ERα blockade with methyl piperidinopyrazole (MPP) led to a significant reduction in both proliferation and differentiation, while ERβ blockade with R,R tetrahydrochrysene (THC) led to an increase in these parameters. This is in keeping with results from mouse knockout models suggesting that unopposed ERβ signalling leads to an inhibition of skeletal growth. Our results are further evidence for the importance of differential ER signalling in regulating chondrogenesis. Future studies examining in vivo effects of these agents are required to extrapolate these findings to a mammalian model."

Although we can't be sure this applies human models.

Estrogen Receptors: How do they signal and what are their targets

Humans do have an ERbeta isoform that matches mice but they also have several other isoforms that have different effects.

"The 530-amino acid (aa)-long human ERβ isoform is currently regarded as the wild-type ERβ (rat and mouse, 549 aa)"

Estrogen Receptor β: Mine Is Longer Than Yours?

According to this next diagram mice and rat ERBeta is longer than human ERBeta.

Localization of estrogen receptors-alpha and -beta and androgen receptor in the human growth plate at different pubertal stages.

"We have investigated the expression of estrogen receptor (ER) alpha, ERbeta and androgen receptor (AR) in biopsies of proximal tibial growth plates obtained during epiphyseal surgery in 16 boys and eight girls. All pubertal stages were represented (Tanner stages 1-5). ERalpha, ERbeta and AR were visualized with immunohistochemistry and the number of receptor-positive cells was counted using an image analysis system. Percent receptor-positive chondrocytes were assessed in the resting, proliferative and hypertrophic zones and evaluated for sex differences and pubertal trends. Both ERalpha- and ERbeta-positive cells were detected at a greater frequency in the resting and proliferative zones than in the hypertrophic zone (64+/-2%, 64+/-2% compared with 38+/-3% for ERalpha, and 63+/-3%, 66+/-3% compared with 53+/-3% for ERbeta), whereas AR was more abundant in the resting (65+/-3%) and hypertrophic zones (58+/-3%) than in the proliferative zone (41+/-3%). No sex difference in the patterns of expression was detected. For ERalpha and AR, the percentage of receptor-positive cells was similar at all Tanner pubertal stages, whereas ERbeta showed a slight decrease in the proliferative zone during pubertal development. In summary, our findings suggest that ERalpha, ERbeta and AR are expressed in the human growth plate throughout pubertal development, with no difference between the sexes."

But how much is the mice isoform of ERBeta that decreases height present in the growth plate?

"To localize ER , a polyclonal rabbit anti-rat ER (06–629; Upstate Biotechnology, Lake Placid, NY, USA), raised against a synthetic peptide corresponding to amino acids 99–116 of human ER (Genbank AF051427) was used."

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