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Objective:To explore the effect of interleukin-1α (IL-1 α) on the senescence of human umbilical vein endothelial cells (HUVECs) and the function of high mobility group protein 1 (HMGB 1).Methods:HUVECs were randomly divided into a control group,a IL-1α group (10 ng/mL IL-1α),a HMGB 1 group (100 ng/mL HMGB 1),and a HMGB 1 +IL-1α group (100 ng/mL of HMGB 1 plus 10 ng/mL of IL-lα).Senescence associated β-galactosidase (SA β-gal) staining was used to assess the number of senescent cells,Western blot were performed to detect the protein levels of silent information regulator 1(SIRT1),and quantitative real-time PCR (qRT-PCR) was used to detect the mRNA levels of p53,p21 and p 16.Restlts:Compared with the control group,the number of SA β-gal positive cells were significantly increased in the IL-1α group (P＜0.05),while the expression of SIRT1 protein significantly decreased (P＜0.01).Compared with the IL-1 α group,the expression of SA β-gal positive cells in the HMGB 1+IL-1α group was decreased and the mRNA levels of p21 and p53 were down-regulated (all P＜0.05),however,there was no statistical significance in the mRNA expression ofp16 (P＞0.05).Conclusion:IL-1α can induce the senescence of HUVECs,and HMGB1 may inhibit IL-1α-induced endothelial cell senescence via p53-p21 pathway.
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Objective To study the effect of baicalein on the skin cell light aging and ERK signaling pathways. Methods Using UVB phototherapy device, of which light intensity was 0.5 mJ/cm2, different exposure time was tried before establishing the best exposure time light aging model. The experiment can be divided into control group, model group and baicalein group (1 × 10-7, 1 × 10-6, 1 × 10-5 mol/L) since the results of preliminary experiments shown that these three concentration of baicalein is the most effective concentration. Using flow cytometry instrument to test cells reactive oxygen species (ROS) content changes of each group. quantitative Real-time RCR (qRT-PCR) and Western blotting were respectively used to detect mRNA and protein expression in each group. Results The best exposure time for 5 min, and cell shrinkage, damage, apoptosis were observed in model group. Compared with model group, cell damage, shrinking phenomenon obviously reduced in 1 × 10-7, 1 × 10-6 mol/L baicalein group, and cell obviously tended to normal form in 1 × 10-5 mol/L baicalein group. Compared with the control group, the ROS content of cells in model group was increased significantly (P < 0.01). Compared with model group, the ROS content was decreased significantly in 1 × 10-7, 1 × 10-6, 1 × 10-5 mol/L baicalein group (P < 0.01); Compared with the control group, the expression of IL-1α and IL-8 mRNA was increased significantly (P < 0.01), and the expression of IL-1-α, IL-8, and p-ERK protein was also increased significantly (P < 0.01) and ERK protein expression quantity remains the same in the model group. In 1 × 10-7, 1 × 10-6, 1 × 10-5 mol/L baicalein group, compared with model group, IL-1α and IL-8 mRNA expression were decreased significantly (P < 0.01), and IL-1-α, IL-8, and p-ERK protein expression were also decreased significantly (P < 0.05, 0.01) and ERK protein expression quantity remains the same. Conclusion Protection of baicalein on UVB induced cell light aging, mainly through reducing ROS content, blocking ERK signaling pathway and inhibiting inflammatory cytokines.
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Objective To observe the mechanism of action of different extracts ofTangwang Mingmu Granules on high glucose induced VEGF and IL-1α gene and protein expressions in vascular endothelial cells.Methods Human umbilical vein endothelial cells EA.hy926 were divided into six groups: blank, high glucose,Tangwang Mingmu Granules, extract 1 (glycoside and flavonoid), extract 2 (organic acid and polysaccharides) and extract 3 (alkaloids) groups. High concentration glucose was used to establish the high glucose model in EA.hy926 cells. The expressions of VEGF and IL-1α mRNA were detected by semi-quantitative RT-PCR. The contents of VEGF and IL-1α protein were tested by ELISA.Results The gene expression and protein levels of VEGF and IL-1α were significantly up-regulated under the high glucose condition (P extract 3> extract 2.Conclusion The action intensity of glycosides and flavonoids, alkaloids, organic acids and polysaccharides on VEGF and IL-1α expression inTangwang Mingmu Granules weakens in sequence.
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ObjectiveTo observe the effect of different extracts ofTangwang Mingmu Granule on hypoxia induced gene expressions in vascular endothelial cells.Methods COCl2 intervention cells were used to copy hypoxia models. Human umbilical vein endothelial cells EA. Hy926 were divided into blank group, hypoxia model group,Tangwang Mingmu Granule group, extract 1 (glycosides and flavonoids) group, extract 2 (orgain acids and polysaccharides) group and extract 3 (alkaloids) group. The changes in gene expressions of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α mRNA were detected by semi-quantitative RT-PCR.ResultsThe gene expression levels of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α were significantly up-regulated under the hypoxic condition (Palkaloids>organic acids and polysaccharides inTangwang Mingmu Granule.
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Objective To provide supportive evidences for the diagnosis of primary major depres-sive disorder (MDD) by analyzing the alterations of proinflammatory cytokines such as IL-1α, IL-6, IL-18, TNF-αas well as the total complement activity (CH50) in serum samples.Methods Hamilton Depression Rating Scale ( HAMD) was used to rate the severity of depression with the patients who were diagnosed as primary MDD.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of IL-1α, IL-6, IL-18 and TNF-αand the CH50 in serum samples from 49 patients and 40 healthy subjects .Results Pa-tients with MDD showed significantly different levels of IL-6, IL-18, TNF-αand CH50 in serum samples as compared with that in healthy subjects (P0.05).There were significant gender differ-ences with levels of IL-1αamong the patients with MDD (P0.05). Conclusion The changes of CH50 and certain proinflammatory cytokines such as IL-6, IL-18 and TNF-αmight be involved in the progression of MDD , suggesting the possibility of using them as indicators for the di-agnosis of MDD.
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Objective:To explore in state of Chlamydia trachomatis persistent infection,the STAT3-TLR2 axis may be activated and mediating abnormal secretion of inflammatory cytokines.Methods: We established acute infection and IFN-γinduced persistent infection model of Ct in HeLa cell.Gene transcription, cytokine secretion and protein expression were detected by using qRT-PCR, ELISA and Western blot respectively in STAT3-TLR2 signaling axis in each Ct infection model.Results: Persistent Ct infections upregulated the transcription of TLR2,significantly increased both the secretion of inflammatory cytokine IL-1αand the expression of STAT3 and TLR2,moreover,enhanced the activation of STAT3 simultaneously.Conclusion: In the Ct persistent infection induced by IFN-γ,the STAT3-TLR2 signaling axis activated significantly in HeLa cell.
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Objective To observe the influence of Huiyang Shengji Ointment and its modified formulae on interleukin-1 (IL-1α) and thromboxane B2 (TXB2) in diabetic rats with chronic skin ulcers, and explore the mechanism for promoting the healing of ulcer.Methods Six out of 160 rats were randomly selected as a blank group, without any further processing. The remaining rats were made diabetic model and randomly divided into five groups after 2 weeks:1 d, 3 d, 5 d, 7 d and 14 d groups. Then, these groups were further divided into normal group (Vaseline ointments), model group (Vaseline ointments), Huiyang Shengji Ointment group (whole formula Ointment), Wenyang Yiqi group (Yiqi group, modified Wenyang Yiqi formula ointments) and Huoxue Shengji group (Huoxue group, modified Huoxue Shengji formula ointments). Normal group and model group were given Vaseline ointments;whole formula group, Yiqi group and Huoxue group were given corresponding ointment. Normal group used the method of skin excision, while other groups used STZ injection-hydrocortisone interference-skin excision-foreign body embedded preparation of composite factors for chronic skin ulcer model. After the appropriate treatment period, the rats were executed and tested for the contents of IL-1α and TXB2 in serum by enzyme-linked immunosorbent assay of five time points.Results In treatment 3 d, the contents of IL-1α in Yiqi group were significantly higher than the blank group, model group, whole formula group and Huoxue group (P<0.05). In treatment 5 d, the contents of IL-1α in whole formula group were significantly higher than the blank group and model group (P<0.05). In treatment 7 d, the contents of IL-1α in each treatment group were significantly higher than blank group and model group (P<0.05), and the whole formula group was higher than the Yiqi group and Huoxue group. In treatment 14 d, the contents of IL-1α in model group and Huoxue group were lower than the blank group (P<0.05). In treatment 3 d, the contents of TXB2 in normal group and the whole formula group were higher than the blank group (P<0.05). In treatment 5 d, the contents of TXB2 in whole formula group were higher than the blank group and the normal group (P<0.05). In treatment 7 d, the contents of TXB2 in Yiqi group were higher than the blank, the model, the whole formula and Huoxue groups (P<0.05). In treatment 14 d, the contents of TXB2 in Huoxue group were higher than the blank and model group (P<0.05), and the contents of TXB2 in the blank group and normal group was lower than those treatment groups (P<0.05).Conclusion Huiyang Shengji Ointment and its modified formulae could promote inflammation, stimulate secretion of inflammatory cytokines, while Yiqi Wenyang ointments played a more active role in promoting inflammation of the early phase of wound surface.
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This study was aimed to investigate effect on the expression of cell inflammatory factor IL-1α, IL-1β and IL-6 of modified San-Jia-San (SJS) decoction on cultured hippocampal neurons injury induced by β-amyloid 25-35 protein (Aβ25-35). Primary cultured hippocampal neurons were divided into the normal group, model group, huperzine A group, low-dose SJS group, middle-dose SJS group, and high-dose SJS group. After selective culture for 7~10 days and the absorption of culture fluid, the blank culture fluid, normal cerebrospinal fluid (normal saline group), huperzine A cerebrospinal fluid, and low-, middle- and high-dose SJS cerebrospinal fluid were added, respectively. The culture fluid was added up to an equivalent medium. The incubation was 2 h under the temperature of 37℃. Ex-cept the normal group, Aβ25-35 dealt with aging (the final concentration was 5 μmol/L) was added to other groups to establish AD cell model. An equivalent amount of culture fluid was added to the normal group. After 24 h of incuba-tion, the cell culture supernatant was collected. Enzyme-linked immunosorbent assay (ELISA) was used in the content detection of IL-1α, IL-1β and IL-6. The results showed that modified SJS decoction can decrease the content of IL-1α, IL-1β and IL-6 in the cell culture supernatant. It was concluded that the modified SJS decoction can effectively inhibit the expression of cell inflammatory factor IL-1α, IL-1β and IL-6. It had certain anti-inflammatory effect to protect hippocampal neurons.
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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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BACKGROUND: It was demonstrated that ultraviolet(UV) B light induces the release of IL-1α in cultured human epithelial cell line and augmentation of GM-CSF production by UVB is reported to be mediated by IL-1α in the murine keratinocyte cell line Pam 212. OBJECTIVE: The purpose of this study was to evaluate the effects of UVB on kinetic profile of IL-1 and GM-CSF mRNA expression and to see whether synthesis of GM-CSF by UVB can be completely inhibited by blocking IL-1α mediated pathway. METHODS: We used a competitive RT-PCR for measuring cytokine gene expression in epithelial cell line after UV radiation. RESULTS: The IL-1α mRNA increased as early as 1h after UV irradiation, and then decreased at 3h after the irradiation. Thereafter, the response of IL-1α mRNA was upregulated with a second peak at 6h after the UV irradiation. However, mRNA for GM-CSF increased at 1h after UV light exposure and anti-IL-1α antibodies could only partially inhibit UV-augmented GM- CSF production. CONCLUSION: UVB induced GM-CSF production seemed to be mainly mediated by UVB induced IL-1α but these results suggest that UVB may also induce GM-CSF production through an IL-1α independent pathway.