To observe the effects of phenylallyl compounds on prostaglandin E2 (PGE2) release in mouse cerebral microvascular endothelial cells (bEnd. 3) stimulated by IL-1beta, and to analyze their structure-activity relationship. Different concentrations of phenylallyl compounds were added separately, and the content of PGE2 induced by IL-1beta in the culture media was measured by ELISA assay. The 50% inhibitory concentration (IC50) of PGE2 was calculated. Studies showed that phenylallyl compounds could affect the PGE2 release differently in bEnd. 3 cells induced by IL-1beta. Close relationships were shown between the inhibitory activities and the location and number of the substituent groups. In conclusion, phenylallyl compounds exhibited inhibitory activities at different extent on PGE2 release in bEnd. 3 cells stimulated by IL-1beta and presented certain structure-activity relationship.
Acrolein ; analogs & derivatives ; isolation & purification ; pharmacology ; Animals ; Brain ; blood supply ; Cells, Cultured ; Cinnamates ; isolation & purification ; pharmacology ; Dinoprostone ; antagonists & inhibitors ; secretion ; Drugs, Chinese Herbal ; chemistry ; Endothelial Cells ; cytology ; metabolism ; Inhibitory Concentration 50 ; Interleukin-1beta ; pharmacology ; Mice ; Microvessels ; cytology ; Propanols ; isolation & purification ; pharmacology ; Structure-Activity Relationship

Cinnamyl alcohol (CAL) is known as an antipyretic, and a recent study showed its vasodilatory activity without explaining the mechanism. Here we demonstrate the vasodilatory effect and the mechanism of action of CAL in rat thoracic aorta. The change of tension in aortic strips treated with CAL was measured in an organ bath system. In addition, vascular strips or human umbilical vein endothelial cells (HUVECs) were used for biochemical experiments such as Western blot and nitrite and cyclic guanosine monophosphate (cGMP) measurements. CAL attenuated the vasoconstriction of phenylephrine (PE, 1 microM)-precontracted aortic strips in an endothelium-dependent manner. CAL-induced vasorelaxation was inhibited by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME; 10(-4) M), methylene blue (MB; 10(-5) M) and 1 H-[1,2,4]-oxadiazolole-[4,3-a] quinoxalin-10one, (ODQ; 10(-6) or 10(-7) M) in the endothelium-intact aortic strips. Atrial natriuretic peptide (ANP; 10(-8) or 10(-9) M) did not affect the vasodilatory effect of CAL. The phosphorylation of endothelial nitric oxide synthase (eNOS) and generation of nitric oxide (NO) were stimulated by CAL treatment in HUVECs and inhibited by treatment with L-NAME. In addition, cGMP and PKG1 activation in aortic strips treated with CAL were also significantly inhibited by L-NAME. Furthermore, CAL relaxed Rho-kinase activator calpeptin-precontracted aortic strips, and the vasodilatory effect of CAL was inhibited by the ATP-sensitive K+ channel inhibitor glibenclamide (Gli; 10(-5) M) and the voltage-dependent K+ channel inhibitor 4-aminopyridine (4-AP; 2 x 10(-4) M). These results suggest that CAL induces vasorelaxation by activating K+ channels via the NO-cGMP-PKG pathway and the inhibition of Rho-kinase.
Animals ; Aorta/drug effects/metabolism/physiology ; Atrial Natriuretic Factor/pharmacology ; Cyclic GMP/*metabolism ; Cyclic GMP-Dependent Protein Kinases/*metabolism ; Dipeptides/pharmacology ; Human Umbilical Vein Endothelial Cells/drug effects/metabolism ; Humans ; Male ; Methylene Blue/pharmacology ; NG-Nitroarginine Methyl Ester/pharmacology ; Nitric Oxide/*metabolism ; Nitric Oxide Synthase/metabolism ; Oxadiazoles/pharmacology ; Phenylephrine/pharmacology ; Phosphorylation ; Potassium Channel Blockers/pharmacology ; Potassium Channels/*agonists ; Propanols/*pharmacology ; Quinoxalines/pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Vasoconstriction/*drug effects ; Vasodilation/drug effects ; rho-Associated Kinases/antagonists & inhibitors/*metabolism

OBJECTIVETo investigate the differentiation of bone marrow derived Thy-1(+)beta(2)M(-) cells (BDTC) into mature and functional liver cells and its mechanism.

METHODSBDTC were cocultured with allyl alcohol (AA)-injured hepatocytes and cultured alone in conditional medium containing HGF and bFGF, respectively. BDTC morphologic transformation was observed with phase-contrast and electron-microscopy. Hepatocyte-specific gene expression in cultured BDTC was identified by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Indocyanine green (ICG) ingestion/excretion and urea, albumin production were carried out to evaluate hepatocyte-related function.

RESULTSSome BDTC derived hepatocyte-like cells with high nuclear to cytoplasmic ratio containing mono- or multi-nuclei and abundant mitochondria, endoplasmic reticulum and glycogenic granules appeared after 7-day culture in both the two culture systems. These cells expressed hepatocyte-specific genes (AFP, OV-6, CK18, etc.), and possessed functions of ICG uptake, albumin production and ammonium metabolism.

CONCLUSIONRat BDTCs could differentiate into mature and functional liver cells in special stimulation systems. Moreover, these differentiations were realized by "transdifferentiation", and might dispense with "cell fusion".

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Fibroblast Growth Factor 2 ; pharmacology ; Gene Expression ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Immunomagnetic Separation ; Keratin-18 ; biosynthesis ; genetics ; Propanols ; pharmacology ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Thy-1 Antigens ; metabolism ; alpha-Fetoproteins ; biosynthesis ; genetics