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Objective To evaluate the changes in the expression of spinal endothelin-1 (ET-1) and its receptors in a mouse model of bone cancer pain (BCP).Methods Ninety-six healthy male SPF C3H/HeN mice,aged 4-6 weeks,weighing 20-25 g,were divided into 2 groups (n=48 each) using a random number table:sham operation group (group S) and BCP group.BCP was produced by injecting α-MEM 20 μl containing 1×104 cells/μ1 NCTC 2472 osteosarcoma cells into the distal medullary cavity of the right femur bone.In group S,t-MEM 20 μl was injected into the distal medullary cavity of the right femur bone.Mechanical paw withdrawal threshold (MWT) and the number of spontaneous flinches (NSF) were measured on 1 day before inoculation (T0) and 4,7,10,14 and 21 days after inoculation (T1-5).Twelve mice of each group were randomly sacrificed at T0,2,4,5,and the lumbar enlargement segments of the spinal cord were harvested to detect the expression of ET-1,endothelin type A receptor and endothelin type B receptor protein and mRNA (using Western blot or real-time polymerase chain reaction).Results The MWT was significantly lower and the NSF was higher at T1 in group S and at T1-5 in group BCP than at T0 (P＜0.05).Compared with group S,the MWT was significantly decreased and the NSF was increased at T2-s,and the expression of ET-1,endothelin type A receptor and endothelin type B receptor protein and mRNA was down-regulated at T2,4,5 in group BCP (P＜0.05).Conclusion The pathophysiological process of BCP is associated with down-regulating the expression of spinal ET-1 and its receptors in mice.
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Inflammatory bowel disease(IBD)is a kind of chronic and non-specific inflammatory disease comprising Crohn’s disease(CD)and ulcerative colitis(UC),the etiology has not yet been clarified. Endothelin-1(ET-1)is an active polypeptide composed of 21 amino acid residues,which can constrict blood vessels by activating voltage-dependent Ca2 + channels in vascular smooth muscle cells. Studies have shown that ET-1 plays an important role in the pathogenesis of IBD. This article reviewed the progress in study on ET-1 in the pathogenesis of IBD.
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Objective To investigate autoantibody against endothelin receptor type A (ENRA-Ab) in patients with systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH).The possibility of autoantibody-mediated pathogenesis in the development of SLE-PAH has also been explored.Methods ENRA-Ab in the serum of SLE-PAH and controls were detected by using a human ETRA epitope peptide-based ELISA.The clinical relevance of ENRA-Ab in SLE-PAH was analyzed.Proliferation of vascular smooth muscle cells (SMCs) and permeability of endothelial cells in vitro under the stimulation of polyclonal ENRA-Ab IgG were assessed.The expressions of PAH-related markers, i.e., 5-HTT, PDGFR-b, VEGF-A and PDGF-B were measured by qPCR.The effect of ENRA-Ab in vivo was also determined in a suboptimaldose monocrotaline-induced model with the assessment of right ventricle hypertrophy index and pathology parameters.Independent t-test, Tukey-Kramer test of variance analysis and Pearson' s correlation analysis were used for statistical analysis.Results ENRA-Abs was presented in a higher occurrence in SLE-PAH (35/85,41％) compared with controls (0/60;0, 13/80, 16％).There was a significant correlation between ENRA-Ab and echocardiograph estimated pulmonary arterial systolic pressure (r=0.392, P=0.002) in SLE-PAH.ENRA-Ab could promote SMCs proliferation, disrupt endothelial barrier and up-regulate PAH-related markers expression,which could be blocked in the presence of ETR antagonist.ENRA-Ab aggravated right ventricle hypertrophy and vascular remodeling in vivo.Conclusion ENRA-Ab is a new biomarker, in SLE-PAH, which may mediate PAH development in SLE.
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Objective To observe the mRNA expressions of endothelin-1(ET-1) and endothelin A/B receptors (ETA/B) in tissue of benign prostatic hyperplasia treated by daily low-dose sildenafil.Methods A total of 32 patients with benign prostatic hyperplasia were randomly divided into two groups:treatment(25 mg sildenafi for 12 weeks,n=16) and control (no drug,n=16) groups.Immunohistochemical staining,ELISA and RT-PCR were used to detect the expression levels of ET-1 protein and ET A/B mRNA,respectively.Results The expressions of ET-1 protein and ET A/B mRNA in prostatic tissue were significantly lower in treatment group than in control group[(53.31±18.56) ng/kg vs.(83.34±31.38) ng/kg,0.356±0.056 vs.0.624±0.083,0.721±0.083 vs.0.933±0.905,t=-3.295,10.715,6.937,all P＜0.001].Conclusions Daily low-dose sildenafil can reduce the expressions of ET-1 and ET A/B receptors mRNA in benign prostatic hyperplasia.
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Endothelin (ET)-1 and its receptors (ETA and ETB receptor) are present in the central nervous system. ET exerts biological effects on gliogenesis and glial cell functions. In order to define a possible mechanism of ETA receptor signaling, the distribution of the ETA receptor in developing oligodendrocytes and the effects of ET-1 on the myelination of oligodendrocytes were examined. ETA receptor immunoreactivity was confined to the perivascular elements of the blood vessels during early postnatal development. However later in development, ETA receptor immunoreactivity was no longer observed in the vessels but became localized to the myelinating oligodendrocytes of the primitive corpus callosum of the white matter, apart from the vessels. ET-1 induced myelin basic protein (MBP) in primary oligodendrocyte precursor cell culture though the ETA receptor and was blocked by an ETA receptor antagonist. In addition, ET-1 evoked the release of Ca2+ which is a central regulator of oligodendrocyte differentiation. Our results provide a link between ET-1 and its ETA receptor and myelination during oligodendrocyte differentiation.
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Animals , Mice , Rats , Brain/pathology , Calcium/metabolism , Calcium Signaling , Cells, Cultured , Endothelin-1/metabolism , Mice, Inbred ICR , Myelin Basic Protein/genetics , Myelin Sheath/physiology , Oligodendroglia/cytology , Rats, Sprague-Dawley , Receptor, Endothelin A/metabolismABSTRACT
Objective To investigate the modulation of ET-3 on the nuclear factor (NF)-κB/Bfl-1 antiapoptotic pathway in a malignant melanoma cell line A375. Methods Flow cytometry was performed to detect the apoptosis in cultured A375 cells after treatment with ET-3 of 100 nmol/L for 24 hours. ET-3 of various concentrations (0, 1, 10, 100 nmol/L) was used to treat some A375 cells with or without the pretreatment with the ETRB antagonist BQ788; after another 24-hour culture, RT-PCR and Western blot were conducted to examine the mRNA expression of Bfl-1 and protein expressions of Bfl-1 and ETRB, respectively. Results The 24-hour treatment with ET-3 of 100 nmol/L significantly reduced the apoptosis rate of A375 cells (F = 10.68, P ＜0.05). The mRNA and protein expressions of Bfl-1 were up-regulated by ET-3 in a concentration dependent manner (both P ＜ 0.01 ), while BQ788 significantly blocked the ET-3-induced up-regulation (F = 420.38,229.49, both P ＜ 0.01 ). The protein expression of pNF-κB in A375 cells was also enhanced by ET-3 of different concentrations (all P ＜ 0.05), but the enhancement was suppressed by BQ788, and there was a significant difference in the protein expression of pNF-κB between cells treated with ET-3 of 100 nmol/L and those treated with the combination of ET-3 of 100 nmol/L and BQ788 (F = 255.46, P ＜ 0.01 ). Conclusion ET-3/ETRB inhibits the apoptosis in A375 cells likely by activating the NF-κB/Bfl-1 anti-apoptotic pathway.
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Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.
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AIM: To investigate the role of endothelin-1 (ET-1), endothelin A receptor (ETAR) in pathophysiology of the lung after mustard intoxication. METHODS: The changes of transcription and expression of ET-1 and ETAR in the lung and their relationship with the tissue damage were studied with immunohistochemistry and RT-PCR. RESULTS: The level of ET-1 in the lung was significantly increased at 2 h, but decreased at 12 h after mustard intoxication. The transcription and expressin of ET-1 and ETAR were significantly increased at the early stage after intoxication. mRNA of endothelin-1 was increased, while mRNA of endothelin A receptor was down-regulated after (12 h.) CONCLUSION: ET-1 and ETAR in lung may play important roles in the pathophysiological process of mustard intoxication. [
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0.05). CO decreased at T2, T3, T4 and T5 compared with that at T0 in both groups (P