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BACKGROUND: Interleukin-1 is an important pro-inflammatory cytokine that has been documented in the regulation of bone inflammation and bone remodeling. A previous study has demonstrated that interleukin-1α can induce apoptosis while inhibiting osteoblast differentiation in MC3T3-E1 cells. OBJECTIVE: To investigate the role and mechanism of interleukin-1α on osteoclast activation and bone loss in mice. METHODS: (1) Cell test: RAW264.7 cells were either treated with interleukin-1α alone or with receptor activator of nuclear factor-κB ligand (RANKL) for 1 and 4 days. Cell viability was tested by cell counting kit-8 assay. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The mRNA and protein levels of osteoclast-specific genes and genes related to nuclear factor-κB pathway and Wnt/β-catenin pathway were tested by real-time fluorescence quantitative PCR, immunofluorescence staining or western blot. Bone marrow-derived macrophages were either treated with interleukin-1α alone or with RANKL and macrophage colony-stimulating factor for 7 days. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The protein levels of osteoclast-specific genes were tested by western blot. (2) Animal test: Twenty-four male C57BL/6J mice (6-8 weeks old) were assigned into two groups at random: control group and test group. Mice were subsequently treated with interleukin-1α solution or PBS by intraperitoneal injection twice a week for 5 weeks. Bone tissues from the femurs were performed with micro-computed tomography analysis and hematoxylin-eosin staining, tartrate resistant acid phosphatase, and immunofluorescence analysis. RESULTS AND CONCLUSION: Cell test: Interleukin-1α alone significantly increased RAW264.7 cell proliferation, but stimulated cell differentiation into osteoclasts in combination with RANKL (P < 0.05). Interleukin-1α significantly increased the expression of osteoclast-related markers and the number of tartrate resistant acid phosphatase-positive multinuclear cells in RAW264.7 cells and bone marrow-derived macrophages in the existence of RANKL or RANKL+macrophage colony-stimulating factor (both P < 0.05). Interleukin-1α was found to significantly enhance the nuclear factor-κB and Wnt/beta-catenin signaling in RAW264.7 cells (P < 0.05). Blocking of nuclear factor-κB or Wnt3 signaling not only reversed the activation of nuclear factor-κB and Wnt3 signaling but also weakened the enhanced expression of osteoclast-specific genes induced by interleukin-1α in RAW264.7 cells (P < 0.05). Animal test: interleukin-1α induced bone loss in mice while also upregulating the expression of osteoclast-specific markers, RANK, TRAF6 and p65, and Wnt3 in vivo (P < 0.05). The findings indicate that interleukin-1α can induce osteoclast activation and bone loss by promoting the nuclear factor-κB and Wnt signaling pathways.
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Objective To examine whether ethanol modulates the intracellular processes involved in the secretion of IL-1α,and then exert a protective effect against coronary heart disease. Methods THP-1 cells in human were cultured for 2-3 generations , and put in PMF for 72 h to induce THP-1 into macrophage. ELISA was applied to detect effects for secretion of IL-1α by LPS, cholesterol and ethanol. In the light of ELISA re-sults, western blot was applied to detect the effects of ethanol on caspase-1 and NLRP3. Results Compared with the control group, the secretion of IL-1α in LPS group and LPS + CHOL group increased. Compared withLPS + CHOL group, the concentration of IL -1α in LPS + CHOL + etha group significantly decrease(P < 0.01). The results of western blot showed that ethanol significantly inhibited caspase-1 and NLRP3 activation. Conclu-sion Ethanol can inhibit the NLRP3 inflammasome activation in macrophages , which may represent a biological pathway underlying the protective effect of moderate alcohol consumption on coronary heart disease.
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Objective To investigate the expressions of inflammatory factors in papillary thyroid carcinoma tissue and their relationships with the clinical pathological characteristics,and to provide an experimental basis for guiding clinical treatment and prognosis.Methods 74 patients with papillary thyroid carcinoma dignosed by puncture were selected as experimental group,and 26 cases of healthy people were used as normal control group.The fasting venous blood samples from the subjects in two groups were collected,and the postoperative specimens from cancer tissue,adjacent normal tissue and the fasting venous blood 7 d after operation of the patients in experimental group were collected again.The interleukin 1α(IL-1α),interleukin 1β(IL-1β)and cyclooxygenase-2 (COX-2)protein expression levels in serum,cancer adjacent normal tissue and cancer tissue were detected by ELISA;the gene and protein expression levels of IL-1α,IL-1βand COX-2 were examined by real-time fluorescent quantitative method and immunohistochemistry;the relationships between their expressions and clinical stages, pathological types,lymph node metastasis of thyroid papillary carcinoma were analyzed.Results The expression levels of serum IL-1α,IL-1βand COX-2 protein of the patients in experimental group were significantly higher than those in normal control group (P<0.01);the gene and protein expression levels of IL-1α,IL-1βand COX-2 in thyroid papillary carcinoma tissue were significantly higher than those in adjacent tissue (P<0.01).There were positive correlations between the expressions of IL-1αand IL-1β,IL-1αand COX-2,IL-1βand COX-2 in thyroid papillary carcinoma tissue (r=0.64,P=0.035;r=0.71,P=0.042;r=0.69,P=0.038).Conclusion The expression levels of IL-1α, IL-1βand COX-2 in thyroid papillary carcinoma tissue are increased, suggesting that inflammation is involved in metastasis and invasion of thyroid papillary carcinoma,its application is expected to become a new means of cancer treatment strategies.
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Objective To explore the preliminary mechanism of mesenchymal stem cells (MSCs) in promoting prostate cancer proliferation in tumor inflammatory microenvironment.Methods From April 2013 to October 2013,MSCs pretreated with inflammatory cytokine IL-1α (MSCs (IL-1α)) and its culture supernatants mixed with RM-1 cells,which origined from C57BL/6 mice,were subcutaneously administered in the armpit area of C57BL/6 or BALB/c mice to establish homologous or heterologous transplant animal mode and to detect the tumor growth.Meanwhile the influence of MSCs on the proliferation of spleen cells was detected in vitro.Results In homologous transplant model,the relative tumor weight of prostate cancer cells prtreated with MSCs and MSCs (IL-1α) and their culture supernatant were (3.4 ± 0.2),(3.3 ±0.2),(4.9±0.5),and (5.2±0.6) g.The results were statistically significant (P＜0.05) compared with the control group (2.4±0.2) g.In heterologous model,the ratio of tumor formation of the pretreated groups were 50％,50％,80％ and 80％,respectively,compared with the control group of 0％.The results were statistically significant (P＜0.05).In proliferation experiments of spleen cells,the number of spleen cell pretreated with IL-1α were significantly lower than that in control group and unpretreated group (P ＜ 0.05).Conclusions MSCs pretreated with IL-1α could effectively promote the growth of prostate cancer cell in vivo.The reason may be due to inflammatory cytokines induce immune suppression of MSCs and then lead to immune escape of cancer cells.
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Objective To investigate the changes and clinical significance of calcitonin gene related peptide (CGRP) and interleu-kin 1α(IL-1α) before and after warm acupuncture therapy in the patients with lumbar disc herniation (LDH ) .Methods Serum CGRP and IL-1αcontents were detected before and after the warm acupuncture therapy in 60 cases of lumbar disc herniation and 15 healthy control cases by ELISA method for conducting the statistical analysis .The changes of the McGill pain scores and serum CGRP and IL-1αconcentrations before and after the warm acupuncture therapy were observed .Results The McGill pain scores af-ter the warm acupuncture therapy in the LDH patients were significantly decreased ,the difference was statistically significant (P<0 .01) ,and serum CGRP and IL-1α concentrations were significantly decreased ,the difference was statistically significant (P<0 .01) .Conclusion CGRP and IL-1αcan be used as the observation index of the effects for the warm acupuncture therapy in trea-ting LDH ,which can provide the basis for the conservative treatment of LDH .
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Objective To study the effect of selective COX-2 inhibitor NS-398 on IL-1α and TNF a expression in HaCaT cells induced by UVA/UVB,and further to explore the mechanism on anti human skin photo damage. Methods The subcuhured HaCaT cells were divided into three groups:simple illuminated group was exposed to UVA/UVB directly,NS-398 interfered group was exposed to UVA/UVB after being treated with NS-398 in different concentrations,and the control group was cultured in normal without any treatment.The expression of IL -1α and TNF-α in supernarant was detected by ELISA kit.Results The level of IL-1α and TNF-α expression in supernatant from simple illuminated group was remarkably higher than that in the control group,NS-398 interfered group showed much lower level than the simple illuminated one,and the expression of IL-1α and TNF-α was dependent on the concentration of NS-398.Conclusions NS-398 can reduce IL-1α and TNF-α expression in HaCaT cells induced by ultraviolet rays,suggesting the possibility of anti human skin photo- damage effect.
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Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before.In this study the expression of PEDF,interleukin-1 α (IL-1α) and -8 (IL-8) in bladder tumours was investigated.Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples.Expression change of the factors was compared with clinicopathological parameters.Correlations between PEDF,IL-1α and IL-8 were analyzed.None of the factors was in relation to gender,tumour occurrence,and size or onset pattem.PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression.PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping.PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049,r=-0.578) as well as in tumour grouping (P=0.033,r=-0.276).Deranged expressional change of PEDF,IL-1 α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.
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BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.
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Background Corneal neovascularization (CNV) is an important cause of visual impairment and graft rejection after allograft corneal transplantation in inflammatory corneal diseases. The mechanisms and therapy relating to CNV are intensely investigated at all times. Objective This study was to evaluate the effect of eicosapentaenoic acid (EPA) on CNV induced by alkali cauterization and its mechanism. Methods The animal models of corneal neovasculation were induced in the right eyes in 72 Sprayue-Dawley rats by putting a piece of 3 mmfilter paper with 1 mol/L NaOH at the center of the cornea for 30 seconds. The rats were then divided randomly into the 0.02 mg EPA treatment group (24 rats) ,0.03 mg EPA treatment group (24 rats) ,model group (24 rats) and normal group (6 rats). EPA of 0.04 ml with doses of 0.02 mg or 0. 03 mg or saline solution of 0. 04 ml was injected subconjunctivally in model rats and immediately after cauterization. The presence of CNV and corneal edema were observed daily by slit lamp biomicroscope. 1,4,7 and 14 days after operation, corneal histopathological examination was performed by hematoxylin and eosin staining. The vascular endothelial cells were stained with CD34 by immunohistochemistry,and the expression of IL-1α,IL-6 mRNA and the nuclear factor-κBp65 ( NF-κBp65 ) proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by Hebei Province( version 1998 ). Results Under the slit lamp, CNV grew slowly from days 2-4 with obvious corneal edema and defect of epithelium. Larger CNV area and less edema were seen from days 7-10. Maximal vessel growth was observed 14 days after injury with thinner vessels in the model group. Histological examination showed that part of the corneal epithelium was damaged;serious corneal edema, more inflammatory cells and a lot of CNV in the stroma were presented in the model group. However, repairing of the corneal epithelium without CNV ,light corneal edema and less inflammatory cells were found in both the 0. 02 mg EPA and 0. 03 mg EPA treatment groups 7 days after alkali cauterization. The relative area of CNV in the 0. 02 mg EPA treatment group was ( 15.80±6.43 )% and ( 11.06±2. 14)% ,and that in the 0. 03 mg EPA treatment group was (16. 10±7.41 )% and (11.06±2. 51 )%, showing significant reduction in comparison with the model group [ (84. 74±7.77)% and (89.63±7.50) % ] 7 days and 14 days after operation ( P＜0. 05 ). Stronger expression of CD34 in the vascular endothelial cells of the cornea stroma was observed in the model group and an absence of CD34 was observed in the EPA-treated groups on the 7th day. RT-PCR revealed that the expression of IL-1α mRNA and IL-6 mRNA was lower in the EPA treatment groups than the model group ( P＜0. 05 ), and Western blot analysis showed that the expression of NF-κB/p65 in the corneas in the EPA treatment groups was significantly lower than that in the model group on the 4th day after operation (P＜0.05).Conclusion Topical application of EPA suppresses CNV induced by alkali burn possibly by inhibiting the expression of NF-κB,IL-1α and IL-6.
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Keratocystic odontogenic tumors have a high level of proliferative activity in epithelial cells and they tend to grow aggressively in the jaw. The tumor dramatically decreases in size by decompression of the intracystic fluid pressure. We herein focused on the roles of interleukin (IL)-1α and demonstrated the biochemical mechanisms of the tumor growth. We found that IL-1α is strongly expressed in the lining epithelial cells of the tumors, and the intracystic fluid levels of IL-1α are significantly higher than the levels of the other inflammatory cytokines of IL-6 and tumor necrosis factor-α (TNF-α). The expression of IL-1α in the epithelial cells decreases after the marsupialization of the tumor. <i>In vitro</i> experiments also reveal that positive pressure enhances the expression of IL-1α in the tumor epithelial cells in culture. IL-1α stimulates the production of matrix metalloproteinase (MMP)-9, and activates the released proMMP-9 by increasing the expression of proMMP-3 and plasminogen activator urokinase (u-PA) in the tumor epithelial cells. In the fibroblasts isolated from the tumors, IL-1α increases the expression of proMMP-1, proMMP-2, and proMMP-3. IL-1α also activates proMMP-2 by inducing the expression of membrane-type 1 matrix metalloproteinase (MT1-MMP) synergistically with type I collagen. Furthermore, IL-1α increases the expression of macrophage colony-stimulating factor (M-CSF) and cyclooxygenase (COX)-2 in the fibroblasts. The COX-2 synthesizes prostaglandin E<sub>2</sub> (PGE<sub>2</sub>), and the secreted PGE<sub>2</sub> stimulates the expression of receptor activator of nuclear factor-κB ligand (RANKL), while neither IL-1α nor PGE<sub>2</sub> affects the expression of osteoprotegerin (OPG) in the fibroblasts. The fibroblasts express Ca<sup>2+</sup>-sensing receptor (CasR) on the cell surface, and extracellular Ca<sup>2+</sup> activates COX-2 expression via the CasR. A strong relationship may thus be present between the intracystic fluid pressure and IL-1α expression in epithelial cells, and the released IL-1α may play a crucial role in the growth of keratocystic odontogenic tumors by stimulating proteolytic enzyme production and osteoclastogenesis.
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The animal models for pharmacologic assessment of drugs against premature delivery are reviewed, which include the measure of spontaneous delivery time between the first and the second pups in term pregnancy rats, the delay in the onset of labor in rats and premature delivery artificially induced by lipopolysaccharide, interleukin-1 α and prostaglandin F2α in mice.