Involvement of Interleukin-1 β/Insulin-Like Growth Factor 1 in Ameliorating Effects of Electroacupuncture on Myocardial Fibrosis Induced by Essential Hypertension / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of electroacupuncture (EA) at Neiguan (PC 6) on myocardial fibrosis in spontaneously hypertensive rats (SHRs), and to explore the contribution of interleukin-1 β (IL-1 β), insulin-like growth factor 1 (IGF-1), and transforming growth factor β 1 (TGF- β 1) to the effects.@*METHODS@#Nine 12-weeks-old Wistar Kyoto (WKY) male rats were employed as the normal group. Twenty-seven SHRs were equally randomized into SHR, SHR+EA, and SHR + sham groups. EA was applied at bilateral PC 6 once a day 30 min per day in 8 consecutive weeks. After 8-weeks EA treatment at PC 6, histopathologic changes of collagen type I (Col I), collagen type 1 (Col 1) and the levels of IGF-1, 1L-1 β, TGF- β 1, matrix metalloproteinase (MMP)-2 and MMP-9 were examined in myocardial tissure respectively.@*RESULTS@#After 8-weeks EA treatment at PC 6, the enhanced myocardial fibrosis in SHRs were characterized by the increased mean fluorescence intensity of Col I and Col 1 in myocardium tissue (P<0.01). All these abnormal alterations above in SHR + EA group was significantly lower compared with the SHR group (P<0.01). Meanwhile, the increased levels of IL-1 β, IGF-1, TGF-β 1 in serum or myocardial tissue of SHRs, diminished MMP 9 mRNA expression in SHRs were also markedly inhibited after 8 weeks of EA treatment (P<0.05 or P<0.01). Furthermore, the contents of IL-1 β, IGF-1, TGF-β 1 in myocardial tissue were positively correlated with the systolic blood pressure and hydroxyproline respectively (P<0.01).@*CONCLUSION@#EA at bilateral PC 6 could ameliorate cardiac fibrosis in SHRs, which might be mediated by regulation of 1L-1 β/IGF-1-TGF- β 1-MMP9 pathway.

Effects of moxibustion on serum levels of β-EP, SP and expression of IL-1β and COX-2 protein in brainstem in rats with migraine / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion at "Baihui" (GV 20) and "Dazhui" (GV 14) at different time points on the serum level of β-endorphin (β-EP), substance P (SP) and expression of interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) protein in brainstem in rats with migraine, and to explore the effect and mechanism of moxibustion in preventing and treating migraine.@*METHODS@#Forty male SD rats were randomly divided into a blank group, a model group, a prevention+treatment (PT) group and a treatment group, 10 rats in each group. Except the blank group, the rats in the remaining groups were injected with nitroglycerin subcutaneously to prepare migraine model. The rats in the PT group were treated with moxibustion 7 days before modeling (once a day) and 30 min after modeling, while the rats in the treatment group were treated with moxibustion 30 min after modeling. The "Baihui" (GV 20) and "Dazhui" (GV 14) were taken for 30 minutes each time. The behavioral scores in each group were observed before and after modeling. After intervention, ELISA method was used to detect the serum level of β-EP and SP; the immunohistochemistry method was used to detect the number of positive cells of IL-1β in brainstem; the Western blot method was used to detect the expression of COX-2 protein in brainstem.@*RESULTS@#Compared with the blank group, the behavioral scores in the model group were increased 0-30 min, 60-90 min and 90-120 min after modeling (P<0.01); compared with the model group, in the treatment group and the PT group, the behavioral scores were decreased 60-90 min and 90-120 min after modeling (P<0.01). Compared with the blank group, in the model group, the serum level of β-EP was decreased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β in brainstem and the expression of COX-2 protein were increased (P<0.01). Compared with the model group, in the PT group and and the treatment group, the serum level of β-EP was increased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β and the expression of COX-2 protein in brainstem were decreased (P<0.01, P<0.05). Compared with the treatment group, in the PT group, the serum level of β-EP was increased and COX-2 protein expression was decreased (P<0.05).@*CONCLUSION@#Moxibustion could effectively relieve migraine. The mechanism may be related to reduce the serum level of SP, IL-1β and COX-2 protein expression in brainstem, and increase the serum level of β-EP, and the optimal effect is observed in the PT group.

Effect of histone H4 on polarization of alveolar macrophages in mice with acute respiratory distress syndrome / 中国职业医学

Objective To investigate the role of histone H4 in the polarization of alveolar macrophages (AM) in lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) in mice. Methods i) The specific pathogen free male C57BL/6 mice were randomly divided into control group and 2, 4, 6 and 8 mg/kg LPS groups, with six mice in each group. The mice in the LPS groups were intratracheally administered LPS according to their respective doses, while the mice in the control group received an equivalent volume of 0.9% saline. After 12 hours, the arterial blood gas was analyzed, and the pulmonary edema and histopathological changes in lung tissues of mice in each group were observed. The level of histone H4 in bronchoalveolar lavage fluid (BALF) of mice was detected using enzyme-linked immunosorbent assay , and mice AMs of the five group were isolated using adherent method. ii) AMs from normal mice were isolated using adherent method and randomly divided into control group, histone H4 injury group, BALF injury group and anti-histone H4 antibody (anti-H4) intervention group. In the histone H4 injury group, AMs were treated with histone H4 at a final concentration of 20 mg/L. In the BALF injury group and anti-H4 intervention group, AMs were treated with 200 μL BALF supernatant from mice intratracheally administered 6 mg/kg body weight LPS, with the latter group treated with 25 mg/L anti-H4 antibody. The control group AMs were treated with phosphate-buffered saline. iii) After 12 hours of stimulation, the cells were collected, and the relative expression of tumor necrosis factor-α (Tnfa), interleukin-1β (Il1b), differentiation antigen 206 (Cd206) and arginase 1 (Arg1) in AMs was detected using real-time quantitative polymerase chain reaction. Results i) Compared with the control group, mice in all four LPS groups exhibited rapid breathing, inflammatory reaction and lung edema in lung tissues, which were aggravated in a dose-dependent manner. The ratio of partial pressure of arterial oxygen to fraction of inspired oxygen in mice decreased with the increase of LPS dose (P<0.05). The wet/dry weight ratio of lung, the level of histone H4 in BALF and the relative expression of Tnfa and Il1b mRNA in AMs increased with the increase of LPS dose (all P<0.05). The mice in the 6 and 8 mg/kg LPS groups developed ARDS. The level of histone H4 in BALF and the relative expression of Tnfa and Il1b mRNA in AMs of mice in 6 and 8 mg/kg LPS groups were higher than those in the other three groups (all P<0.05). ii) The relative expression of Tnfa and Il1b mRNA increased (both P<0.05), and the relative expression of Cd206 and Arg1 mRNA decreased (both P<0.05) in AMs of histone H4 injury group and BALF injury group compared with the control group. Compared with BALF injury group, the relative mRNA expression of Tnfa and Il1b in AMs of anti-H4 intervention group decreased (both P<0.05), while the relative expression of Arg1 mRNA increased (P<0.05). Conclusion LPS can induce a dose-dependent increase in histone H4 levels in BALF in mice. Histone H4 drives the development of ARDS by activating AMs to M1 polarization. Antagonizing histone H4 to interfere with AM polarization to M1 could be a target for the treatment of ARDS.

JNK inhibitor SP600125 inhibits interleukin-1β induced pyroptosis in βTC-6 cells / 中华内分泌代谢杂志

Objective:To investigate whether interleukin(IL)-1β is involved in pyroptosis which leads to mouse islet β cell line βTC-6 cell damage, and to explore the role of JNK inhibitor SP600125 in inhibiting IL-1β induced βTC-6 cell pyroptosis.Methods:βTC-6 cell line and mouse islets were incubated with IL-1β for 48 h or intervened with both JNK inhibitor SP600125 and IL-1R antagonist IL-1Ra, then GSDMD expression and β cell pyroptosis morphology were detected by immunofluorescence staining of GSDMD and DAPI. The expression levels of Gsdmd, IL-1β and IL-18 mRNAs were detected by real time fluorescence PCR, and apoptosis was examined by Annexin-V/7-AAD staining combined with flow cytometry.Results:βTC-6 cell pyroptotic body was significantly increased in the IL-1β treated group compared with the control group, and the expressions of pyroptosis related genes Gsdmd, IL-1β, and IL-18 mRNA were significantly higher( P<0.05), and apoptosis was increased, suggesting that IL-1β effectively induced the βTC-6 cell pyroptosis, IL-1Ra prevented IL-1β induced βTC-6 cell pyroptosis. In the presence of JNK inhibitor SP600125, IL-1β treatment failed to induce the expressions of Gsdmd and IL-18 mRNA, markers of pyroptosis, and reduced the rate of apoptosis, indicating that SP600125 suppressed IL-1β induced βTC-6 cell pyroptosis. Conclusion:Pyroptosis is one of the mechanisms of βTC-6 cell impairment caused by IL-1β, and SP600125, a JNK inhibitor, can block the IL-1β induced pyroptosis pathway and has a potential role in inhibiting βTC-6 cell pyroptosis.

Relationship between interleukin-1β and silicosis development / 中国基层医药

Silicosis is a diffuse pulmonary fibrosis disease caused by occupational exposure to silica, which is one of the occupational diseases with high incidence in developing countries. Up to now, there is no definite drug to relieve or reverse the lung injury caused by silicosis, so it is very important to prevent, diagnose and treat pulmonary fibrosis as soon as possible. Studies have shown that a chronic inflammatory environment contributes to pulmonary fibrosis to a certain extent. Interleukin-1β is a cytokine that increases the number of inflammatory factors in the microenvironment in the immune response and plays a key role in inflammatory reaction. Therefore, the release of interleukin-1β is of great significance in the pathogenesis of silicosis. This paper aims to systematically expound the development course of silicosis, the signal pathway of interleukin-1β production, and the relationship between them.

Mechanism of Qingre Lishi Huazhuo Method（Modified Simiaosan） on Chronic Gouty Arthritis of Dampness-heat Obstruction Syndrome Based on NLRP3/IL-1β Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the clinical efficacy of the Qingre Lishi Huazhuo method on patients with chronic gouty arthritis of dampness-heat obstruction syndrome and the effect on nucleotide-binding oligomerization domain-like receptor 3（NLRP3）/interleukin-1β （IL-1β） signaling pathway to preliminarily explore its mechanism. MethodSixty patients with chronic gouty arthritis of dampness-heat obstruction syndrome were enrolled and divided into a treatment group （30 cases） and a control group （30 cases） according to the random number table method. Thirty people were assigned to the healthy group. Patients in the control group were treated with oral Febuxostat， while those in the treatment group were treated with modified Simiaosan combined with Febuxostat. Treatment lasted four weeks. The general clinical data， traditional Chinese medicine （TCM） syndrome scores， serum uric acid （UA）， serum creatinine （SCr）， blood urea nitrogen （BUN）， fasting blood glucose （FPG）， low-density lipoprotein （LDL）， triglyceride （TG）， total cholesterol （TC）， erythrocyte sedimentation rate （ESR）， and C-reactive protein （CRP） of patients were recorded. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of IL-1β，TNF-α， and IL-6，and the levels of NLRP3，cysteinyl aspartate-specific protease-1 （Caspase-1）， and apoptosis-associated speck-like protein containing a CARD （ASC） were detected by Western blot. ResultBefore treatment， the levels of body mass index （BMI）， systolic blood pressure （SBP），diastolic blood pressure （DBP），UA，SCr，BUN，FPG，LDL，TG，and TC in both groups significantly increased （P<0.05，P<0.01），and the levels of HDL significantly decreased as compared with those in the healthy group（P<0.05）. Additionally， the levels of IL-1β， TNF-α， and IL-6 in both groups significantly increased before treatment （P<0.01）. Compared with the results before treatment， patients in the two groups had significant reductions in tube pain， joint tenderness， joint swelling，joint fever， activity disorders， body fatigue， sliminess， bitter mouth， yellow and red urine， and tongue manifestation scores （P<0.05，P<0.01）. Compared with patients in the control group after treatment， those in the treatment group had a significant decrease in joint fever， body fatigue， sliminess， bitter mouth，sticky stool，yellow and red urine， tongue manifestation score， and pulse score （P<0.05）. The total effective rate in the treatment group was 80.0% （24/30）， higher than 56.7% （17/30）in the control group（χ2=11.916，P<0.05）. Compared with the results before treatment， BMI， SBP， DBP， UA， SCr， BUN， FPG， LDL， TG， TC， ESR，CRP， IL-1β， TNF-α， IL-6 levels， and VAS score in both groups significantly decreased （P<0.05，P<0.01）. Compared with patients in the control group after treatment， those in the treatment group had decreased DBP，ESR， IL-1β levels， and VAS score （P<0.05）. Western blot results showed that before treatment， the protein expression of NLRP3， Caspase-1， and ASC in peripheral blood mononuclear cells （PBMCs） of patients in both groups were higher than those in the healthy group （P<0.01）. Compared with the results before treatment， the protein expression of NLRP3， Caspase-1， and ASC in PBMCs in patients of both groups after treatment decreased （P<0.05，P<0.01）. Compared with the control group after treatment， the treatment group showed decreased expression levels of NLRP3 and Caspase-1（P<0.05）. ConclusionThe Qingre Lishi Huazhuo method can effectively improve the clinical symptoms and reduce inflammation of chronic gouty arthritis of dampness-heat obstruction syndrome with good safety. The mechanism may be related to the inhibition of the NLRP3/IL-1β signaling pathway.

Therapeutic effects of Guchisan combined with orthodontics therapy on chronic periodontitis with syndrome of excessive stomach fire in 33 patients / 中国基层医药

Objective:To investigate the effects of Guchisan combined with orthodontics therapy on serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels. Methods:Sixty-six patients with chronic periodontitis who received treatment in Zhejiang Xin'an International Hospital between July 2013 and July 2019 were included in this study. They were randomly divided into control group ( n = 33) and treatment group ( n = 33). Both control and treatment groups received conventional interventions. The control group was treated with orthodontics therapy. The treatment group was treated with Guchisan (0.5 g once, twice a day, 2-3 minutes once) based on orthodontics therapy for 1 month. Gingival index, probing depth, attachment loss, plaque index, gingival bleeding index, and latency to symptom improvement were compared between the two groups. Clinical efficacy was compared between the two groups. Serum IL-1β and TNF-α levels were measured in each group. Results:After treatment, gingival index [(0.51 ± 0.06) vs. (1.12 ± 0.15), t = 21.69], probing depth [(2.01 ± 0.22) mm vs. (2.67 ± 0.30) mm, t = 6.50], attachment loss [(1.10 ± 0.13) mm vs. (1.70 ± 0.18) mm, t = 9.90], plaque index [(0.47 ± 0.05) vs. (1.08 ± 0.13), t = 25.57], and gingival bleeding index [(0.58 ± 0.07) vs. (1.09 ± 0.13), t = 19.84] were significantly lower than those in the control group (all P < 0.01). Latency to tooth loosening [(8.81 ± 0.96) days vs. (13.03 ± 1.62) days, t = 15.01], halitosis [(3.04 ± 0.33) days vs. (7.03 ± 0.76) days, t = 27.66], and swollen gums [(2.21 ± 0.24) days vs. (4.55 ± 0.51) days, t = 23.85] were significantly shorter than those in the control group (all P < 0.01). Total response rate in the treatment group was significantly higher than that in the control group (93.94% vs. 69.70%, χ2 = 4.99, P = 0.025). After treatment, serum levels of TNF-α [(4.01 ± 0.44) ng/L vs. (5.31 ± 0.58) ng/L, t = 10.26] and IL-1β [(16.04 ± 1.97) mg/L vs. (18.30 ± 2.55) mg/L, t = 4.03] in the treatment group were significantly lower than those in the control group (both P < 0.01). Conclusion:Based on conventional treatment, Guchisan combined with orthodontics therapy for treatment of chronic periodontitis with syndrome of excessive stomach fire can greatly improve symptoms of periodontal disease and enhance clinical efficacy. Lowering serum TNF-α and IL-1β levels may be one of effective action ways of this combined therapy.

Correlation between hand-arm vibration disease and vasoactive factors / 环境与职业医学

Background Hand-arm vibration disease (HAVD) is a chronic progressive disease caused by long-term exposure to hand-transmitted vibration, but the mechanism by which vibration affects peripheral vascular function of fingers is not completely clear. Objective To study the association between vasoactive factors and HAVD, and to screen specific indicators for its early diagnosis and prevention. Methods Judgmental sampling method was used to select workers with (HAVD group) and without HAVD (vibration contact group), and non-hand-transmitted vibration operation workers (control group), with 60 workers in each group. The levels of leukotriene B4 (LTB4), vascular endothelial growth factor (VEGF), 5-hydroxy tryptamine (5-HT), interleukin-1β (IL-1β), and calcitonin gene-related peptide (CGRP) in plasma of the three groups were measured by enzyme-linked immunosorbent assay. The association between vasoactive factors and HAVD was analyzed using logistic regression, and the diagnostic HAVD indicators were screened by receiver operator characteristic (ROC) curve of a multivariate model indicator \begin{document}$ \widehat{Y} $\end{document}. Results The hand symptom rates of the HAVD group, the vibration contact group, and the control group were 26.7%, 66.7%, and 96.7% respectively, with a significant difference (P<0.05). The levels of LTB4, 5-HT, IL-1β, and CGRP in the HAVD group were the highest followed by the vibration contact group, and lowest levels were in the control group (P<0.05). There was no significant difference in the VEGF level among the three groups (P>0.05). The logistic regression results showed that higher levels of LTB4 (OR=1.048, 95%CI: 1.022-1.076), 5-HT (OR=1.011, 95%CI: 1.004-1.018), IL-1β (OR=1.148, 95%CI: 1.071-1.230), and CGRP (OR=1.055, 95%CI: 1.008-1.104) were associated with a higher risk of HAVD (P<0.05). The order of the potential indicators' area under the ROC curve from high to low was:\begin{document}$ \widehat{Y} $\end{document} (0.969) > IL-1β (0.907) > LTB4 (0.876) > 5-HT (0.858) > CGRP (0.836). Conclusion The expression levels of LTB4, 5-HT, IL-1β, and CGRP are altered with occupational exposure in hand-transmitted vibration operations and may be associated with HAVD; VEGF is not found to be associated with HAVD. The accuracy of early screening for HAVD can be improved by combining the monitoring of various biochemical indicators.

Effects of paraquat on proliferation and neurogenesis of neural stem cells by promoting secretion of IL-1β by microglia / 环境与职业医学

Background Paraquat (PQ) is a widely used herbicide that exerts neurotoxicity. The effects of PQ on neural stem cells (NSCs) through microglia mediated neuroinflammation remain limitedly studied. Objective To investigate the effects of PQ on the proliferation and neurogenesis of NSCs through neuroinflammation mediated by microglia. Methods Microglial cell lines (BV2 cells) and primary NSCs were used. BV2 cells were exposed to 0, 1, 3.3, 10, 33, and 100 μmol·L−1 of PQ for 6 h followed by viability assessment. The highest PQ concentration that had no effect on cell viability was selected as the final exposure concentration (33 μmol·L−1). To exclude the direct effect of PQ on NSCs, after the BV2 cells were cultured in complete medium containing 33 μmol·L−1 PQ for 6 h, the BV2 culture medium was replaced by NSCs complete medium without PQ for 24 h. The concentration of interleukin-1β (IL-1β) in supernatant was detected by enzyme-linked immune sorbent assay. Besides, in order to detect the effects of IL-1β on NSCs proliferation and neurogenesis, NSCs isolated from hippocampus of adult mice were cultured in the supernatant obtained above and divided into four groups: control supernatant + control antibody, control supernatant + IL-1β neutralizing antibody (10 ng·mL−1), PQ supernatant + control antibody, PQ supernatant + IL-1β neutralizing antibody (10 ng·mL−1). Proportion of Ki67-positive NSCs was detected by flow cytometry (FCS) and immunofluorescence after 24 h culture, and neurogenesis was detected by FCS and immunofluorescence after 3-7 d of culture. Results The IL-1β concentration in the supernatant of BV2 cells was significantly increased after the 33 μmol·L−1 PQ exposure compared with the control group (t=3.020, P<0.05). After the NSCs were cultured with the supernatant of PQ-treated BV2 cells, the proportion of Ki67-positive NSCs (t=9.129, P<0.01) and the proportion of newborn neurons (t=4.638, P<0.01) were significantly decreased compared to the control group. After neutralizing IL-1β, the proportion of Ki67-positive NSCs (t=22.05, P<0.01) and the proportion of newborn neurons (t=11.09, P<0.01) were significantly higher than those in the un-neutralized group. The results of immunofluorescence detection also showed that after neutralizing IL-1β secreted by 33 μmol·L−1 PQ-treated BV2 cells, the number of Ki67-positive NSCs and the number of newborn neurons were significantly higher than those in the un-neutralized group. Conclusion The secretion of IL-1β by microglia is increased after PQ treatment, resulting in a decrease in the proliferation and neurogenesis of NSCs. These results suggest that neuroinflammation is involved in NSCs damage caused by PQ.

The effects of extracorporeal shock wave treatment on the expression of TGF-β1 and IL-1β in the cartilage of an osteaoarthritic knee / 中华物理医学与康复杂志

Objective:To seek any effect of extracorporeal shockwave treatment on the expression of transforming growth factor-β1 (TGF-β1) and interleukin-1β (IL-1β) in the cartilage tissue of rabbits with knee osteoarthritis (OA), and its therapeutic mechanism.Methods:Fifty female New Zealand rabbits were randomly divided into a normal control group, a model group, and three shockwave groups A, B and C, each of 10. Except for the normal control group, an OA model was established in the other groups using Hulth′s method. The shockwave groups were given 2000 shocks in each weekly session over 4 weeks. The energy flow density in group A was 0.05mJ/mm 2; in B it was 0.11mJ/mm 2 and in C 0.22mJ/mm 2. The normal control and model groups were not shocked. All the rabbits were then sacrificed and their right knee cartilage tissue was sampled to observe any pathological changes and assign improved Mankin scores. Immunohistochemistry was used to count the number of TGF-β1 and IL-1β-positive cells in the cartilage. Western blotting and real-time fluorescence quantitative polymerase chain reactions were employed to determine the protein and mRNA expression of IL-1β and TGF-β1. Results:Compared with the normal group, degeneration of articular cartilage was observed in the model group. The average Mankin′s score of the model group was significantly higher than that of the normal control group. The average expression of TGF-β1 and IL-1β protein and mRNA in the model group had increased significantly compared with the normal control group. The average Mankin′s scores of the shock wave groups were all significantly lower than the model group′s average. Group C′s average expression levels of TGF-β1 and IL-1β protein and mRNA were significantly lower than the model group′s averages.Conclusions:Extracorporeal shockwave therapy can reduce the expression of TGF-β1 and IL-1β in the cartilage of an arthritic knee, at least in rabbits. Its therapeutic effect is positively correlated with the density of the energy flow, suggesting that shock waves may reduce the expression of inflammatory factor IL-1β by regulating the expression of TGF-β1. They should be applied in the prevention and treatment of osteoarthritis.

Interleukin-1β mediates the effect of macrophage androgen receptor on hyperphosphate-induced vascular smooth muscle cell calcification / 中华肾脏病杂志

Objective:To investigate whether it is by regulating interleukin 1β ( IL-1β) gene expression that androgen receptor (AR) in macrophages affects hyperphosphate-induced vascular smooth muscle cell calcification. Methods:The chromatin immunoprecipitation (ChIP) experiment was used to determine whether AR was bound to the androgen receptor element (ARE) sequence of IL-1β promoter in THP-1 cells. Whether the AR regulated IL-1β gene expression was detected by luciferase assay experiments. AR of THP-1 cells was silenced and transfected by lentivirus with vector or shRNA. Flow cytometry was used to select positive transfected cells THP-1ARsc (control) and THP-1ARsi (AR silencing) with fluorescent markers. Western blotting was used to detect AR protein levels of THP-1ARsc (control) and THP-1ARsi cells (AR silencing in monocytes). Macrophages MФARsc (control) or MФARsi (AR silencing) were induced by 50 ng/ml phorbol ester. Enzyme-linked immunosorbent assay was used to detect IL-1β expression levels of MФARsc or MФARsi conditioned medium. The human aortic smooth muscle cells (HASMC) were cultured in MФARsc or MФARsi conditioned medium with phosphate (2.5 mmol/L final concentration of sodium dihydrogen phosphate), and Alizarin red S staining was used to analyze HASMC calcification degree. Western blotting was used to detect the expression levels of RUNX2 (osteoblast marker) and SM22α (HASMC marker), and neutralization assay was performed to test IL-1β-mediating effect of macrophages AR on HASMC calcification. Results:AR was bound to ARE sequence of IL-1β promoter and regulated IL-1β gene expression. The expression level of IL-1β protein in conditioned medium of MФARsi cells decreased significantly compared to MФARsc cells ( P<0.001). Compared with MФARsc conditioned medium group, HASMC calcium deposition in MФARsi conditioned medium group decreased significantly, RUNX2 protein decreased and SM22α protein increased (all P<0.05). The degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group decreased than that in the MФARsc conditioned medium+IgG antibody group significantly, and the degree of HASMC calcification in the MФARsc conditioned medium+IL-1β antibody group decreased significantly than that in the MФARsc conditioned medium+IgG antibody group; while the degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group and MФARsi conditioned medium+IL-1β antibody group decreased than that in the MФARsc conditioned medium+IL-1β antibody group (all P<0.05). Conclusions:Macrophage AR regulates IL-1β expression by binding to ARE sequence within IL-1β promoter, and IL-1β mediates the effect of macrophage AR on hyperphosphate-induced HASMC calcification.

High glucose promotes the release of IL-1β and IL-18 from placental trophoblast by activating NLRP3 inflammasome / 中华内分泌代谢杂志

Objective:To investigate the effect of high glucose on the release of interleukin (IL)-1β and IL-18 in placental trophoblast by activating NLRP3 inflammasome.Methods:Gestational diabetes mellitus(GDM) placentas and control placentas were collected and the expression levels of NLRP3 and Caspase-1 were determined. Human placental trophoblast HTR-8/SVneo were cultured and divided into control group(5.5 mmol/L glucose), high glucose group(25 mmol/L glucose), DMSO+ high glucose group, and Ac-YVAD-cmk(NLRP3 inflammasome inhibitor)+ high glucose group. The expression levels of NLRP3 and Caspase-1 in cells as well as the contents of IL-1β and IL-18 in the medium were determined.Results:The expression levels of NLRP3 and Caspase-1 in GDM placenta were higher than those in control placenta( P<0.05) and positively correlated with homeostasis model assessment of insulin resistant index(HOMA-IR) and fasting insulin. The expression levels of NLRP3 and Caspase-1 in HTR-8/SVneo cells and the secretion levels of IL-1β and IL-18 in high glucose group were higher than those in control group( P<0.05). Ac-YVAD-cmk significantly suppressed high glucose-stimulated IL-1β and IL-18 secretion( P<0.05). Conclusion:High glucose promotes the release of IL-1β and IL-18 from placental trophoblast via activating NLRP3 inflammasome.

NLRP3 inflammasome and its role in age-related macular degeneration / 中华实验眼科杂志

Age-related macular degeneration (AMD), the leading cause of central vision loss among people aged 50 years and older, is one of the major eye diseases causing blindness in the world.Clinically, advanced AMD is divided into two types, non-exudative AMD with manifestation of geographic atrophy and exudative AMD with manifestation of choroidal neovascularization.The pathogenesis of AMD is complex, and the para-inflammation is recognized as an important risk factor.Nucleotide-binding oligomerization domain like receptors 3 (NLRP3) inflammasome is a cytoplasmic pattern recognition receptor and is expressed in several kings of cells, including retinal pigment epithelium (RPE) cells, microglial cells, Müller glia cells and retinal vascular endothelial cells.Recent studies have suggested that NLRP3 inflammasome plays an important role in the pathophysiology of both non-exudative and exudative AMD.The role of the NLRP3 inflammasome and its effector cytokines interleukin (IL)-1β and IL-18 in AMD were reviewed in this article to provide guidance on future prevention and therapy of AMD.

Research progresses on NLRP3 inflammasomes-induced anti-tumor immunity / 药学学报

More and more studies have shown that NOD-like receptor protein 3 (NLRP3) inflammasome has become the regulatory factor of inflammatory response and protective immunity, and the assembly and activation of NLRP3 inflammasomes are closely related to the anti-tumor immunity effect. Depending on the cell type and stimuli, activation of the NLRP3 inflammasome can induce immune cells to become polarized, hyperactive, or pyroptotic, releasing interleukin (IL)-1β and IL-18, which leads to cascade immune or inflammatory responses, and its role in tumor immunity has received extensive attention. Here, we review the mechanisms of the NLRP3 inflammasome enhancing CD8+ T cells-mediated anti-tumor immunity by inducing the pyroptosis of tumor cell, the pyroptosis or hyperactive state of dendritic cells (DCs), and the pyroptosis or polarization of the macrophages. Different anti-tumor immune roles of NLRP3 inflammasome activation in tumor cells and immune cells provide new directions for future research and may influence the development of next-generation immunotherapy.

Mechanism of Chaihu Qinggantang in Intervening NLRP3/IL-1β Pathway to Treat Granulomatous Lobular Mastitis in Rat Model / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Chaihu Qinggantang （CHQGT） in the treatment of granulomatous lobular mastitis （GLM） in the rat model. MethodSixty female rats were divided into a normal group， a model group， a prednisolone group （0.001 8 g·kg-1）， and three CHQGT low-dose， medium-dose， and high-dose groups （4.5， 8.9， 17.8 g·kg-1）. The tissue homogenates mixed with GLM lesion tissue and Fritner's reagent were used for modeling. After modeling， the treatment groups were given corresponding treatment factors， and the normal group and the model group were given the equal volume of normal saline. The changes in mammary gland of rats were observed after 14 d. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in breast samples. The mRNA expressions of NOD-like receptor thermal protein domain associated protein 3 （NLRP3） inflammasome， Caspase-1， and interleukin-1β （IL-1β） were detected by real-time quantitative fluorescence polymerase chain reaction （Real-time PCR）. The protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 were detected by Western bolt. ResultAs compared with the normal group， the breasts of rats in the model group were obviously swelling， and mammary gland inflammation index was significantly increased （P<0.01）. Pathological changes included the formation of granuloma centered on the lobule of mammary gland with a large number of inflammatory cells such as lymphocytes and plasma cells. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL18 in the model group were significantly increased （P<0.01）. Compared with the model group， the treatment groups improved breast swelling， and the CHQGT medium and high-dose groups and the prednisolone group reduced inflammation index to some extent after treatment （P<0.05， P<0.01）. The inflammation degree of mammary gland was significantly improved， and inflammatory cells such as macrophages， lymphocytes， and plasma cells were reduced to varying degrees in pathological aspects. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 in the CHQGT high-dose group and the prednisolone group were significantly down-regulated （P<0.05， P<0.01）. ConclusionCHQGT inhibits inflammation and treats GLM in rats. The mechanism is possibly related to the inhibition of NLRP3/IL-1β signaling pathway， which provides a new target for the prevention and treatment of GLM by Qingxiao method.

Tumor-DerivedIL-1β Promotes Adhesion of Vascular Endothelial Cells with Hepatoma Cells Through NF-κB p65 Signaling Pathway / 华中科技大学学报(医学版)

Objective To elucidate the effect of tumor-derived interleukin-1 beta(IL-1β)on adhesion of vascular endothelial cells(HUVECs)with hepatoma cells and the potential molecular mechanisms. Methods HepG2 cell line stably expressing IL-1β(HepG2/IL-1β)and GFP(HepG2/GFP)was established by lentivirus transfection,and the IL-1β in cell culture supernatant was determined by ELISA. Conditioned medium(CM)containing IL-1β(IL-1β-CM)of HepG2 cells was used as a source for tumor-derived IL-1β,and was controlled with CM from HepG2/GFP cells(Ctrl-CM). HUVECs stimulated with 10 ng/mLIL-1β(HUVECs+IL-1β-CM)were used to observe their adhesion ability with HepG2/IL-1βand HepG2/GFP cells,and HUVECs+CtrlCM served as control. The mRNA expression levels of E-selectin(CD62E) ,ICAM-1(CD54)and VCAM-1(CD106)in HUVECs were detected byqRT-PCR. Cell surface expression levels of CD62Eand CD54 were detected by flow cytometry.IL-1β induced signaling pathways in HUVECs were analyzed through Western blotting. Specific inhibitors were usedto block each signaling pathway,and the expression of adhesion molecules as well as the adhesion of vascular endothelial cells with hepatoma cells were subsequently monitored. Results Tumor-derived IL-1β significantly enhanced the adhesion of HUVECs with HepG2 cells and upregulated the expression levels of CD62E and CD54 mRNAin HUVECs cells. The surface CD62E was temporarily upregulated 4 h after IL-1β stimulation,while the expression of CD54 protein showed a continuous upregulated pattern,which lasted from 4 hto 24 h.IL-1β mainly activated NF-κB p65 and p38 MAPK signaling pathways in HUVECs,and NF-κB p65 was proved to be located at upstream,regulating the activation of p38 MAPK pathway. Targeted blocking of NF-κB p65 pathway by specific inhibitor significantly downregulated the expression of CD62E and CD54 on surface of HUVECs and inhibited their adhesionability with HepG2cells. Conclusion Tumor-derived IL-1β promotes adhesion of HUVECs with hepatoma cells through upregulation of CD62E and CD54 expression by activating NF-κB p65 signaling pathway in HUVECs. Tumor-derivedIL-1β might be involved in invasion and metastasis of hepatoma cells through vascular system.

Expression features of cytochrome C oxidase subunit Ⅰ, BCL2 interacting protein 3 and interleukin-1β in MRL/lpr lupus mice liver tissue / 中华风湿病学杂志

Objective:To explore the expression features of cytochrome C oxidase subunit Ⅰ (MT-CO1), BCL2 interacting protein 3 (BNIP3) and interleukin (IL)-1β in the liver of MRL/lpr lupus mice.Methods:The mRNA and protein levels of MT-CO1, BNIP3, IL-1β, p16 and p21 in lupus mice and control mice were detected by polymerase chain reaction (PCR) and Western blot, the IL-1β expression site were detected by hematoxylin and eosin (HE) staining and immunohistochemical method, and themalondialdehyde (MDA) was detected by colorimetry. Hepatocytes and macrophages were stimulated with lipopolysaccharide (LPS), while hepatocytes were also cultured with supernatants obtained after macrophages stimulated with LPS, and the mRNA and protein levels of MT-CO1, BNIP3 and LC3B, as well as p16 and p21 expression, were determined by qPCR and Western blot. The expression of mitochondrial reactive oxygen species (mtROS) was detected by immunofluorescence. One way Analysis of Variance (ANOVA) was used to compare the mean of each group, and LSD method was used to compare the means of multiple samples, and Tamhane's T2 method was used to compare the means of multiple samples when the variance was uniform. Results:The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the liver tissue of the lupus group (0.14±0.04; 0.16±0.05) were significantly lower than those of the control group (0.11±0.04; 0.16±0.06), and the differences were statistically significant ( t=7.16, P<0.001; t=4.54, P<0.001). The expression levels of IL-1β, p16 and p21 in the lupus group (2.06±0.69; 0.37±0.14; 0.16±0.06) were significantly higher than those of the control group (0.23±0.06; 0.25±0.08; 0.11±0.04) ( t=9.58, P<0.001; t=24.35, P<0.001; t=22.36, P<0.001). The results of Western blot were consistent with those of PCR. HE staining showed lymphocyte infiltration in the liver tissue of lupus mice, and immunohistochemistry showed IL-1β in the liver tissue of lupus mice. The positive cells were mainly concentrated in the sinusoids, and the expression of hepatic parenchymal cells was not rearkable. The content of MDA in liver tissue of the lupus group (0.19±0.10) was higher than that of the control group (0.17±0.09), and the difference was statistically significant ( t=4.33, P=0.005). LPS directly stimulated AML12 hepatocytes (0.069±0.028; 0.17±0.07). The PCR results showed that compared with the control group (0.176±0.072; 0.08±0.03), the expression of MT-CO1, and BNIP3 were not significantly different ( t=1.01, P=0.337; t=0.88, P=0.399). The expression of IL-1β was significantly higher when incubated with the supernatants of LPS stimulated macrophages (0.28±0.09) compared than that of the control group (0.15±0.05) ( t=28.26, P<0.001). The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the LPS stimulated group (0.046±0.026; 0.17±0.05) were significantly lower than those in the control group (0.143±0.083; 0.18±0.06), and the differences were statistically significant ( t=7.52, P<0.001; t=4.24, P<0.001), The expression of p16 and p21 in LPS stimulated group (0.29±0.09; 0.27±0.09) were significantly higher than those in the control group (0.18±0.06; 0.22±0.07) ( t=13.54, P<0.001; t=8.69, P<0.001). The results of Western blot were consistent with those of PCR. Immunofluorescence showed that the fluorescence intensity of mtROS in LPS stimulated group (0.25±0.10) was higher than that in the control group (0.08±0.03), and the difference was statistically significant ( t= 4.86, P<0.001). Conclusion:Immune-mediated inflammation in the liver tissue of lupus mice can stimulate liver parenchymal cells to cause intracellular mitochondrial dysfunction. However, the mechanism of liver organ damage in lupus mice is not limited to the immune-mediated inflammation of immune active cells, but also include parenchymal cell mitochondrial dysfunction.

Block nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome to alleviate non-alcoholic steatohepatitis and liver fibrosis / 中华消化杂志

Objective:To explore the role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in non-alcoholic steatohepatitis (NASH).Methods:The liver tissue samples of 24 patients admitted the Second Affiliated Hospital of Fujian Medical University were selected, including 12 NASH samples from liver biopsy and 12 normal liver tissues from the margin of hepatic hemangioma. The expression of NLRP3, apoptosis-associated speck-like protein (ASC), caspase-1, interleukin (IL)-1β and the content of triglyceride (TG) were detected. Wild-type and NLRP3 -/- C57BL/6 mice were fed with normal diet or methionine-choline deficient diet (MCD) for 8 weeks. The wild-type mice were divided into MCC950 NASH, 0.9% sodium chloride (NaCl) NASH, MCC950 and 0.9% NaCl group, 8 mice in each group, and were fed with MCD diet and treated with MCC950, fed with MCD diet and treated with 0.9% NaCl, fed with normal diet and treated with MCC950, and fed with normal diet and treated with 0.9% NaCl respectively for eight weeks. After eight weeks, the pathologic changes of liver tissues were observed with hematoxylin-eosin staining and immunohistochemistry. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), free fatty acid (FFA), IL-1β and TG in serum were determined by enzyme linked immunosorbent assay. The expression levels of NLRP3, ASC, caspase-1 and IL-1β in liver tissues were examined by Western blotting and real-time quantitative polymerase chain reaction. Primary Kupffer cells were isolated and cultured from the livers of wild-type and NLRP3 -/- mice and divided into control group and palmitic acid group. The expression levels of related proteins in the supernatant of cells culture were detected by Western blotting. Independent sample t test and one-way ANOVA were used for statistical analysis. Results:The expression levels of NLRP3, ASC, caspase-1, IL-1β and the content of TG of the liver tissues of the NASH patients were all higher than those of healthy control group (all P<0.05). The formation of steatohepatitis in hepatocyte of MCD-fed mice was more obvious than that of nomal diet-fed mice, with more hepatocyte ballooning and inflammatory cell infiltration. The expression of NLRP3, ASC, caspase-1, IL-1β, caspase-1 activity and the content of TG in liver tissue of NASH mice were all higher than those of normal diet-fed group (all P<0.05); and serum levels of ALT, AST, IL-1β, and the content of FAA were all higher than those of normal diet-fed group (all P<0.05). The serum levels of ALT, AST, IL-1β and IL-18 of NLRP3 -/- NASH mice were all lower than those of wild-type NASH mice (all P<0.05). The serum level of ALT, the expression of ASC, caspase-1 and IL-1β in liver tissues, and the degrees of liver fibrosis of wild-type MCC950 NASH group were all lower than those of 0.9% NaCl NASH group (all P<0.05). The expression of NLRP3, ASC, caspase-1, caspase-1 activity, and secretion of IL-1β and IL-18 in Kupffer cells from wild-type mouse treated with palmitic acid were all higher than those of the negative control group (all P<0.05). However, the changes of the above indicators in Kupffer cells from NLRP3 -/- mouse were not affected by palmitic acid treatment. Conclusion:NLRP3 blockade can significantly alleviate the liver injury and fibrosis in NASH mice and prevent the development of NASH.

Effects of mechanical vibration on micoRNA-214-3p and serum interleukin-1β in ovariectomized rats with a fracture / 中华物理医学与康复杂志

Objective:To observe the effect of mechanical vibration on the expression of microRNA-214-3p and serum interleukin-1β (IL-1β) in the broken ends of fractured bones in ovariectomized rats.Methods:Thirty 3-month-old female Wistar rats were randomly divided into a control group, a model group, and a vibration group, each of 10. An operation of ovariotomy was performed in those of the model and vibration groups to establish osteoporosis model. Five months later a model of mid femur fracture was made with animals in all the groups. Five days after their fracture, the vibration group received 20 minutes of whole-body vibration treatment at 35Hz for 5 days a week, while the control group and the model group received natural rearing without any additional intervention At 2 and 6 weeks after the operation, the Lane-Sandhu X-ray scoring system was used to evaluate the quality of fracture healing, and reverse-transcription polymerase chain reactions and enzyme-linked immunosorbent assays were used to detect miR-214-3p in the fractured bones and the serum levels of IL-1β.Results:At 2 weeks and 6 weeks after the operation the average growth score of the broken ends in the model group was significantly lower than that of the control group, while that of the vibration group was significantly higher than the model group′s average. Compared with the control group at the same time point, the average miR-214-3p content of the model and vibration groups was significantly higher 2 and 6 weeks after the surgery. Compared with the model group, the average level of miR-214-3p of the fractured ends of the vibration group was significantly lower at 6 weeks. Two and six weeks after the surgery, the average IL-1β of the model group was significantly higher than the control group′s average, and that of the vibration group was significantly lower.Conclusion:Mechanical vibration can promote osteoporotic fracture healing by inhibiting the expression of miR-214-3p and reducing the level of IL-1β at the broken ends of fractured bones.

The effect of radiofrequency heating on the expression of interleukin-1 beta and tumor necrosis factor alpha in osteoarthritic synovial membranes of the knee / 中华物理医学与康复杂志

Objective:To explore the effect of radiofrequency heating on the morphology of articular cartilage in the knee and the expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the synovium using a rabbit model of knee osteoarthritis (OA).Methods:Fifty-four male rabbits had OA induced in their right hind limbs using the modified Hulth method. They were then randomly divided into a model group, a cervus and cucumis polypeptide (CCP) group and a radiofrequency thermotherapy (RT) group, each of 18. The CCP group was injected with deer melon peptide intramuscularly every day, while the RT group was given daily radiofrequency hyperthermia treatment at 36.5-38.5 ℃. The model group was not provided with any special treatment. On the 6th, 12th and 18th day of the treatment, 6 rabbits in each group were sacrificed to resect the right femur′s medial condyle cartilage. The morphological characteristics of the cartilage were evaluated using modified Mankins scoring, while the contents of lL-1B and TNF-a in the synovial membrane were detected using enzyme-linked immuno-sorbent assays.Results:The average Mankins scores and the levels of IL-1β and TNF-α decreased significantly at each time point, and significant differences were observed among the three groups. In the RT group the average Mankins score as well as the IL-1β and TNF-α levels decreased significantly with time throughout the experiment.Conclusions:Radiofrequency hyperthermia is superior to the injection of deer melon polypeptide in knee osteoarthritis, at least in rabbits. The therapeutic mechanism may be related to the control of IL-1β and TNF- α levels in the synovial membrane.