Humans ; Liver ; cytology ; metabolism ; Liver Diseases ; metabolism ; pathology ; therapy ; Receptors, Angiotensin ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, Endothelin ; metabolism ; Receptors, Vasopressin ; metabolism

Endothelin derived from the endothelial cells of microvessel is a potent vasoactive peptide, which has various physiologic actions in many internal organs. The fact that endothelin receptors are present on the osteoblastic cells suggests that endothelin play a role in bone metabolism. This study was done to study the effect of endothelin-1 on osteoblast and the combined effect of dexamethasone and endothelin-1 on osteoblast. Human osteoblasts isolated from ilium were cultured in DME/F12 medium, and divided into 5 groups; Group 1 (control), Group 2(10(-7)M endothelin-1), Group 3(10(-7)M endothelin-1+1:2500 monoclonal antibody), Group 4(10(-7)M dexamethasone+10(-7)M endothelin-1), and Group 5(10(-7)M dexamethasone). [3H]-thymidine uptake in groups was 23373.2+/-2722.4 cpm/well, significantly higher than that of control (P<0.05), and the increase was blocked by the addition of monoclonal antibody to endothelin(group 3). [3H]-thymidine uptake in groups adding steroid with or without endothelin was significantly lower than that of other groups (P<0.05). Group 2 showed marked increase in type I procollagen mRNA compared with other groups, but group 3 and 4 showed no significant effect on the expression of type I procollagen mRNA. In histochemical staining for alkaline phosphatase activity, the cells in groups with steroid were strongly positive in staining, large in size and looked well differentiated. Osteocalcin synthesis was also increased in groups with steroid treatment compared with other groups. This study demonstrated that endothelin-1 stimulated DNA synthesis and the expression of type I procollagen mRNA in human osteoblasts, and inhibited alkaline phosphatase activity, but had no significant effect on osteocalcin. Dexamethasone stimulated alkaline phosphatase activity and osteocalcin synthesis, and inhibited DNA synthesis but had no significant effect on the expression of type I procollagen mRNA. Dexamethasone masked the effect of endothelin-1 on human osteoblastic cells, and the effect of dexamethasone was predominant in the group of a combination of endothelin-1 and dexamethasone.
Alkaline Phosphatase ; Collagen Type I ; Dexamethasone ; DNA ; Endothelial Cells ; Endothelin-1 ; Endothelins* ; Humans ; Ilium ; Masks ; Metabolism ; Microvessels ; Osteoblasts* ; Osteocalcin ; Receptors, Endothelin ; RNA, Messenger

The purpose of the present study was to assess the correlation that likely exists among increased portal pressure (Pp), portal blood flow quantity (Qp) and ETA and ETB receptor mRNA expression in human cirrhosis. In situ hybridization and reverse-transcription polymerase chain reactions (RT-PCR) were performed to determined the expression of ETA and ETB receptor mRNA in liver tissues from traumatic subjects (n = 10) and cirrhotic patients (n = 15) in whom hepatic hemodynamic values were measured. The expression of the two transcripts was significantly higher in liver samples of cirrhotic patients than in those obtained from traumatic subjects. It has shown that ETA receptor mRNA predominantly located in hepatic stellate cells (HSCs) and vascular smooth muscle cells of intrahepatic arteries and portal veins, ETB receptor mRNA in HSCs, sinusoidal endothelial cells and Kuppfer cells. There was a highly significant direct relationship between ETA and ETB receptor mRNA and Pp and Qp in cirrhotic patients. It suggests that liver paracrine endothelin system may be overactivated in human cirrhosis accompanied with increased expression of ETA and ETB receptor mRNA which may play an important role in the pathogenesis and maintenance of splanchnic hyperdynamics.
Gene Expression ; Hemodynamic Processes ; Hypertension, Portal/metabolism ; Liver Cirrhosis/genetics ; Liver Cirrhosis/*metabolism ; Portal Vein/*physiopathology ; Receptors, Endothelin/genetics ; Receptors, Endothelin/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; *Splanchnic Circulation/physiology

The purpose of the present study was to assess the correlation that likely exists among increased portal pressure (Pp), portal blood flow quantity (Qp) and ETA and ETB receptor mRNA expression in human cirrhosis. In situ hybridization and reverse-transcription polymerase chain reactions (RT-PCR) were performed to determined the expression of ETA and ETB receptor mRNA in liver tissues from traumatic subjects (n = 10) and cirrhotic patients (n = 15) in whom hepatic hemodynamic values were measured. The expression of the two transcripts was significantly higher in liver samples of cirrhotic patients than in those obtained from traumatic subjects. It has shown that ETA receptor mRNA predominantly located in hepatic stellate cells (HSCs) and vascular smooth muscle cells of intrahepatic arteries and portal veins, ETB receptor mRNA in HSCs, sinusoidal endothelial cells and Kuppfer cells. There was a highly significant direct relationship between ETA and ETB receptor mRNA and Pp and Qp in cirrhotic patients. It suggests that liver paracrine endothelin system may be overactivated in human cirrhosis accompanied with increased expression of ETA and ETB receptor mRNA which may play an important role in the pathogenesis and maintenance of splanchnic hyperdynamics.
Female ; Gene Expression ; Hemodynamics ; Humans ; Hypertension, Portal ; metabolism ; Liver Cirrhosis ; genetics ; metabolism ; Male ; Portal Vein ; physiopathology ; Receptors, Endothelin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Splanchnic Circulation ; physiology

OBJECTIVETo investigate the effect of endothelin and its receptors on ouabain-induced hypertension in rats.

METHODSMale Sprague-Dawley (SD) rats were treated with ouabain or saline for 6 weeks and their systolic blood pressure (SBP) were recorded weekly. At the end of 2, 4 and 6 weeks, respectively, the plasma and left ventricle endothelin contents were measured by radio-immunoassay, and real-time quantitative RT-PCR was employed to determine the mRNA level of endothelin type A receptor (ETAR) and type B receptor (ETBR) in the left ventricle, and the protein expressions of ETAR and ETBR were examined by immuno-histochemistry.

RESULTSAfter 4 weeks of intraperitoneal ouabain injection, the mean SBP in ouabain group increased till reaching a level significantly higher than that in the control group after 6 weeks (P<0.001). The plasma and left ventricle endothelin contents were significantly increased after 2 weeks of ouabain injection (P<0.01), and similarly, increased ETAR mRNA was observed. After 4 weeks of treatment, ETAR mRNA was increased continuously and the protein expression of ETAR upregulated in ouabain group as compared with the control group. The transcription and protein expression of ETBR were not altered by ouabain treatment.

CONCLUSIONBefore detectable blood pressure elevation occurs, endothelin concentration and ETAR can be already upregulated in ouabain-induced hypertensive rats, suggesting that endothelin might be involved in the cardiovascular effects of ouabain via an action on ETAR.

Animals ; Endothelins ; biosynthesis ; blood ; genetics ; Hypertension ; chemically induced ; genetics ; metabolism ; Immunohistochemistry ; Male ; Myocardium ; metabolism ; Ouabain ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, Endothelin A ; genetics ; metabolism ; Receptor, Endothelin B ; genetics ; metabolism ; Receptors, Endothelin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effects of the epidermal growth factor on the mRNA expression of endothelin-1 and its receptors (ET(A)R, ET(B)R) in hormone refractory prostate cancer (HRPC) PC-3 cell lines.

METHODSPC-3 cells were cultured in vitro. After the treatment with EGF, the mRNA expressions of endothelin-1, ET(A)R and ET(B)R were detected by RT-PCR in PC-3 cell lines. The levels of the mRNA expression of endothelin-1 and its receptors were examined at different time points by RT-PCR.

RESULTSThe expressions of endothelin-1 and ET(A)R mRNA but not the mRNA expression of ET(B)R was observed in PC-3 cell lines. After 24 hours of treatment with EGF, the expressions of endothelin-1 and ET(A)R in PC-3 cell lines were both up-regulated and there was significant difference (P < 0.05) between the experimental and control groups. Different expression levels of endothelin-1 and ET(A)R mRNA were noted at different time points of EGF intervention, up-regulated with the increase of treatment time, and with significant difference (P < 0.05).

CONCLUSIONEGF can up-regulate the mRNA expressions of endothelin-1 and ET(A)R in PC-3 cell lines and play a great role in prostate cancer progression, which may offer a substructure of molecular biology for the treatment of HRPC.

Cell Line, Tumor ; Endothelin-1 ; genetics ; Epidermal Growth Factor ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Receptor, Endothelin A ; genetics ; Receptor, Endothelin B ; genetics ; Receptors, Endothelin ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the role of advanced glycation end products (AGEs) and their receptors (RAGE) in the pathogenesis of diabetic mellitus erectile dysfunction (DMED) and the effects of AGEs and RAGE on the activity of endothelin-1 (ET-1) in rat cavernosum.

METHODSForty male Sprague-Dawley rats were taken at random to construct 2 groups of diabetes mellitus (DM) models of equal number, one given free access to water and the other administered aminoguanidine hydrochloride (DM + AG) in water at the dose of 1 g/L. Another 20 male SD rats were equally divided into a normal control and an AG control group. After 8 weeks, the cavernosum tissues were harvested from all groups of rats, part of the isolated penile tissues homogenated to detect the content of AGE-peptide (AGE-P) and the activity of ET-1, and the AGEs and RAGE in the rest of the penile tissues analyzed by immunohisto- chemical assay.

RESULTSCompared with the normal controls, the expressions of AGEs and RAGE, the content of AGE-P and the activity of ET-1 in the cavernosum tissues were significantly high in the DM group (P < 0.05), while the administration of AG to the DM rats reversed the above results. No significant difference was observed between the normal control and AG control groups in any of the data (P > 0.05).

CONCLUSIONIn DM conditions, the joint effect of AGEs and RAGE may elevate the activity of ET-1 in rat cavernosum and thus promote the development of DMED.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; prevention & control ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; administration & dosage ; Glycation End Products, Advanced ; antagonists & inhibitors ; metabolism ; Guanidines ; administration & dosage ; Male ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; metabolism

OBJECTIVE: To explore the therapeutic effects and mechanisms of the combination of traditional Chinese medicine and western medicine on patients with peptic ulcers. METHODS: One hundred and twenty patients were randomly divided into 6 groups. Another 10 patients as the control group were confirmed with no peptic ulcers by endoscope, but had digestive tract symptoms. The clinical effects were compared among each group after the one month treatment. RESULTS: The clinical effects of the combination of Jianweiyuyang granules and ranitidine capsules were better than those of western medicine, with improvement in symptoms and syndrome (P < 0.01 to 0.05), but there was not significant difference with the rate of ulcer healing and the Hp clearance among the combination of Jianweiyuyang granules and ranitidine capsules, Jianweiyuyang granules, and ranitidine capsules (P > 0.05). The combination of Jianweiyuyang granules and ranitidine capsules could significantly upregulate the expression of MUCSAC mRNA (P < 0.01), while downregulate the expression of ETAR mRNA (P < 0.01). CONCLUSION There is obvious advantage in treating peptic ulcers by the combination of Jianweiyuyang granules and ranitidine capsules, and its mechanisms may be to protect the gastric mucosal barrier by up-regulating the expression of MUCSAC mRNA and to improve the gastric mucosal blood flow by down-regulating the expression of ETAR mRNA.
Adult ; Capsules ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Endothelin-1 ; biosynthesis ; genetics ; Female ; Gastric Mucosa ; metabolism ; Humans ; Male ; Middle Aged ; Mucin 5AC ; Mucins ; biosynthesis ; genetics ; Peptic Ulcer ; drug therapy ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Ranitidine ; therapeutic use ; Receptors, Endothelin ; biosynthesis ; genetics

OBJECTIVETo explore the role of tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis of hepatorenal syndrome.

METHODSBy isolated perfused kidney technique, rat kidneys were perfused at a constant flow. Changes in perfusion pressure (mmHg) were consecutively measured with multi-functional physiology recorder. After TNF-alpha or heparin treated 90 minutes, the perfusion pressure stimulated by endothelin-1 (ET-1) was detected.

RESULTSTNF-alpha and heparin didn't modify the baseline perfusion pressure. When ET-1 was added at 2 nmol/L, the perfusion pressures increased to (47+/-9) mmHg, (97+/-36) mm Hg and (11+/-6) mm Hg in control, TNF-alpha and heparin (10mg/L) treated group, respectively, which were different among the three groups (t>or=3.811, P<0.01). No pathological damages were found in kidney tissues from all the groups after being stained with hematoxylin/eosin.

CONCLUSIONTNF-alpha plays an important role in the pathogenesis of hepatorenal syndrome by promoting renal vasoconstriction.

Animals ; Calcium Channels ; metabolism ; Endothelin-1 ; pharmacology ; Heparin ; pharmacology ; Hepatorenal Syndrome ; etiology ; metabolism ; In Vitro Techniques ; Inositol 1,4,5-Trisphosphate Receptors ; Kidney ; blood supply ; Male ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Renal Artery ; physiopathology ; Tumor Necrosis Factor-alpha ; analysis ; pharmacology ; Vasoconstriction ; drug effects

To systematically clarify the effects of apolipoprotein E (aopE) and low-density lipoprotein receptor (LDLR) gene mutant on hyperlipidemia, vascular inflammation impairment and pathogenesis of atherosclerosis (AS), total RNA was isolated from fresh aortas of young apoE/LDLR double knockout (apoE(-/-)/LDLR(-/-)) and wild type (WT) mice using TRIzol reagent. Then RNA was reversely transcribed to first-strand cDNA by reverse transcriptase for reverse transcription polymerase chain reaction (RT-PCR) and real-time RT-PCR. Primer pairs were designed using primer design software according to the gene sequences available in GenBank. β-actin was used as an internal control. Then RT-PCR assay was used to analyze the expression patterns of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), nuclear factor-κB (NF-κB), granulocyte-macrophage colony-stimulating factor (GM-CSF), CD36, endothelin-1 (ET-1), toll-like receptor 2 (TLR2), monocyte chemoattractant protein-1 (MCP-1), vascular adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and platelet-derived growth factor-α (PDGF-α). SYBR Green quantitative real-time RT-PCR was used to validate gene expressions identified by RT-PCR. Blood samples were taken from the retro-orbital venous plexus, and serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were measured by using biochemical techniques. Serum concentrations of circulating TNF-α, IL-1β and oxidized LDL (ox-LDL) were determined by ELISA. Frozen sections of aortic sinus were stained with Sudan IV to visualize intimal fatty lesions. The results showed that the relative expressions of IL-1β, GM-CSF, ET-1, TLR2, CD36, MCP-1, ICAM-1 and VCAM-1 in apoE(-/-)/LDLR(-/-) mice at the age of 1 month were higher than those in age-matched WT mice (P<0.05, P<0.01), respectively. The expressions of PDGF-α and TNF-α in apoE(-/-)/LDLR(-/-) mice at the age of 2 months were up-regulated compared to those in age-matched WT mice (P<0.05). All the expressions of target genes continued to be up-regulated (P<0.05, P<0.01) except that ET-1 expression at the age of 2 months, TLR2, VCAM-1 and ICAM-1 expressions at the age of 3 months were down-regulated to that in WT mice. NF-κB expression had no significant changes between two genotype mice at different ages. All the gene expressions kept unchanged in WT mice at different ages, except that IL-1b expressions were slightly up-regulated at the ages of 2 and 3 months. Serum levels of TC, TG, LDL, HDL, TNF-α, IL-1β and ox-LDL in apoE(-/-)/LDLR(-/-) mice at different ages were higher than those in age-matched WT mice (P<0.05, P<0.01), and were increasing with age. Primary atherosclerotic lesions were observed in 1-month old apoE(-/-)/LDLR(-/-) mice and were progressing with age. There were no lesions observed in all WT mice at different ages. The data suggest that hyperlipidemia due to apoE and LDLR gene mutant may stimulate the temporal expressions of AS-related genes and contribute to primary atherogenetic lesions and vascular inflammation impairment.
Animals ; Aorta ; metabolism ; Apolipoproteins E ; genetics ; Atherosclerosis ; genetics ; CD36 Antigens ; metabolism ; Chemokine CCL2 ; metabolism ; Endothelin-1 ; metabolism ; Gene Expression ; Granulocyte-Macrophage Colony-Stimulating Factor ; metabolism ; Hyperlipidemias ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; blood ; metabolism ; Lipoproteins, LDL ; blood ; Mice ; Mice, Knockout ; NF-kappa B ; metabolism ; Platelet-Derived Growth Factor ; Receptors, LDL ; genetics ; Toll-Like Receptor 2 ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism