ObjectiveTo investigate the association between IL-1β gene-511C/T polymorphisms and the risk of chronic obstructive pulmonary disease (COPD). MethodsSuch databases as PubMed, EMbase, CNKI, CBM, VIP and WanFang Data were searched for the studies on the association between IL-1β gene-511C/T polymorphisms and the risk of COPD up to May 2014. According to inclusion and exclusion criteria, two reviewers independently screened literature, extracted data, and assessed methodological quality of included studies. Then meta-analysis was performed using RevMan 5.0 software. ResultsA total of 10 case-control studies from 9 articles involving 1 171 cases and 1 268 controls were included. The results of meta-analysis showed that, no significant association was found between IL-1β gene-511C/T polymorphisms and the risk of COPD:TT+CT vs. CC:OR=1.06, 95%CI 0.66 to 1.70, P=0.82; TT vs. CT+CC:OR=0.87, 95%CI 0.60 to 1.26, P=0.32; TT vs. CC:OR=0.95, 95%CI 0.51 to 1.75, P=0.86; CT vs. CC:OR=1.10, 95%CI 0.71 to 1.70, P=0.15; T vs. C:OR=0.97, 95%CI=0.72 to 1.30, P=0.84. The results of subgroup analysis by ethnicity showed that, no significant association was found between IL-1β gene-511C/T polymorphisms and the risk of COPD among Caucasians and Asians. ConclusionIL-1β gene-511C/T polymorphisms might not contribute to the risk of COPD.
Objective To study the expression of NLRP3 inflammasome and its downstream inflammatory factors in patients with chronic obstructive pulmonary disease (COPD) and healthy controls, and to reveal the effect and significance of NLRP3 inflammasome in the pathogenesis of COPD. Methods Forty patients with acute exacerbation COPD (AECOPD) who were hospitalized from November 2016 to May 2017 were recruited in the AECOPD group, and recruited in the stable COPD group when they entered the stable stage. Forty healthy individuals were recruited in the control group. General information and peripheral blood were collected from each subject. The levels of NLRP3 mRNA and caspase-1 mRNA in peripheral blood mononuclear cells were measured by real-time PCR. The levels of IL-18 and IL-1β were measured by enzyme-linked immunosorbent assay. Results The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the stable COPD group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/ml vs. 1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/ml, all P<0.05] . The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the control group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/mlvs. 1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. The levels of NLRP3 mRNA, IL-18 and IL-1β were significantly higher in the stable COPD group than the control group [1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/mlvs. (1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. Correlation analysis showed that the plasma IL-18 level was positive correlated with leukocyte count and neutrophil percentage in the AECOPD group (r=0.372, P<0.05;r=0.386, P<0.05). The expression of NLRP3 mRNA in the AECOPD group and stable COPD group were positively correlated with the CAT score (r=0.387, P<0.05;r=0.399, P<0.05) . Conclusion NLRP3 inflammasome is involved in the inflammatory response in COPD patients.
Objective To discuss the relationship between the changes of hepatic blood flow detected by usingspectral Doppler ultrasound and serum TNF- α and IL-1 β levels after liver ischemia/reperfusion （I/R） of rat. Methods The hepatic ischemia 15 min and reperfusion models were established by using pringle method. The hepatic blood flow of hepatic artery and portal vein at 1， 6， and 24 hours after liver I/R were detected by using spectral Doppler ultrasound， the total blood flow volume （FV） was calculated， and the serum TNF- α and IL-1 β levels at each time point were detected. The correlation between the TNF-α， IL-1 β， and FV were analyzed. Results The FV at 1 hour and 6 hours after reperfusion in I/R group were less than those in sham operation （SO） group 〔（52.08±11.88） mL/min vs. （85.32±29.85） mL/min and （44.69±8.75）mL/min vs. （81.41±28.67） mL/min， P＜0.05〕. The FV at 24 hours after operation or reperfusion of 2 groups was no significant differences （P＞0.05）. The serum content of TNF-α at 1 hour after reperfusion in I/R group was higher than that in SO group 〔（310.52±39.83）pg/mL vs. （240.74±31.65）pg/mL， P＜0.05〕. The serum contents of TNF-α at 6 and 24 hours after operation or reperfusion of 2 groups were no significant differences （P＞0.05）. The serum contents of IL-1β at 1 hour and 6 hours in I/R group were higher than those in SO group 〔（38.08±3.73） pg/mLvs. （22.03±0.79） pg/mL and （27.44±6.11） pg/mL vs. （21.78±0.71） pg/mL， P＜0.05〕. The serum content of IL-1β at 24 hours after operation or reperfusion of 2 groups was no significant differences （P＞0.05）. There was a negative correlation between the FV and TNF-α or IL-1β （r=-0.43， P＜0.05；r=-0.46， P＜0.05）. Conclusions Spectral Doppler ultrasound can observe the changes of hepatic blood flow and evaluate the hepatic microcirculation indirectly. The hepatic blood flow after liver I/R decreases and it may be related to over expression of TNF-α and IL-1β.
ObjectiveTo investigate the relationship between alumina ceramic particles and aseptic loosening of the joint prosthesis and the effect of lanthanum chloride on the secretion of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) of macrophage RAW264.7 induced by alumina ceramic particles. MethodsRAW264.7 cells were cultured in vitro and divided randomly into 4 groups according to different culture solutions:blank control group (group A),1 mg/mL alumina ceramic particles (group B),1 mg/mL alumina ceramic particles and 10 μmol/L lanthanum chloride (group C),and 10 μmol/L lanthanum chloride (group D).The cell growth was detected by MTT,and ELISA,RT-PCR,and Western blot were used to test the expressions of IL-1β,TNF-α,and nuclear factor κB (NF-κB). ResultsThere was no significant difference in cell growth among all groups by MTT (F=2.180,P=0.142).RT-PCR results showed that the expressions of IL-1β,TNF-α,and NF-κB mRNA in group B were significantly higher than those in the other 3 groups (P<0.05); the expressions in group D were significantly lower than those in group A (P<0.05).ELISA results showed that the contents of IL-1β and TNF-α in group B were significantly higher than those in the other 3 groups (P<0.05); the contents in group D were significantly lower than those in group A (P<0.05).Western blot analysis revealed that the expression of NF-κB protein in group B was significantly higher than that in the other 3 groups (P<0.05). ConclusionAlumina ceramic particles can stimulate the secretion of IL-1β and TNF-α of macrophage,and lanthanum chloride can inhibit the secretion of IL-1β and TNF-α of macrophage.
ObjectiveTo isolate and identify the cartilage progenitor cells (CPCs) from normal cartilage, and to explore the influence of interleukin 1β (IL-1β) in different concentrations on its chondrogenesis. MethodsCPCs were isolated from normal cartilage of adult New Zealand white rabbit with the fibronectin adhesion assay;the cell phenotype was identified;and the cloning and differentiation of CPCs were observed. CPCs were incubated with H-DMEM in group A, with chondrogenic induced medium in group B, with chondrogenic induced medium+0.1 ng/mL IL-1β in group C and chondrogenic induced medium+1.0 ng/mL IL-1β in group D for 3 weeks. The histology, biochemistry, and real-time fluorescence quantitative PCR were performed to observe the effect of IL-1β on the chondrgenic differentiation. ResultsThe CPCs from normal cartilage expressed positively stem cell phenotype, which have similar ability of cloning and differentiation to stem cells. The cell pellets in groups C and D were significantly smaller than those in group B, and cell showed hypertrophic morphology change. There were more expressions of collagen type Ⅱ and collagen type X in group B than in group A, in group B than in groups C and D, and in group C than group D with Safranin O staining. The biochemistry results showed that collagen type Ⅱ content, glycosaminoglycan (GAG) content, and the ratio of GAG/DNA were significantly lower in groups C and D than in group B (P<0.05), and in group D than in group C (P<0.05);but the DNA content was significantly higher in groups C and D than in group B (P<0.05), and no significant difference between groups C and D (P>0.05). The real-time fluorescence quantitative PCR results showed that the relative mRNA expressions of collagen type Ⅱ, collagen type X, and Sox-9 were significantly lower in groups C and D than in group B (P<0.05), and in group D than in group C (P<0.05), but the relative mRNA expressions of Runx-2 and matrix metalloproteinase 13 were significantly higher in groups C and D than in group B (P<0.05), and in group D than in group C (P<0.05). ConclusionThere are CPCs having the character of stem cells in normal cartilage, and they have the capability of cloning and potential differentiation. IL-1β can inhibit the chondrogenesis of CPCs, and possibly promote the osteogenic differentiation.
Objective To access the protective effects of glucosamine hydrochloride capsules (OTL) on articular cartilage in osteoarthritis of rabbit. Methods Thirty-six New Zealand white rabbits were divided randomly into three groups (n=12): sham group (group A), anterior cruciate l igament transection (ACLT)/normal sal ine group (group B), and ACLT/ OTL group (group C). Rabbits in groups B, C received ACLT on the right knee. Rabbits in group A were not given ACLT ascontrol. Group C received a daily administration of OTL at a dose of 150 mg/kg of body weight for 12 weeks; in contrast, group B received normal sal ine at the same dose. All rabbits were sacrificed after 12 weeks. The right femoral condyle were removed and observed at pathologic changes with HE staining and graded by Mankin’s scale, the expression level of transforming growth factor β1 (TGF-β1) and interleukin 1β (IL-1β) were detected by immunohistochemical staining. Results All rabbits survived at the end of experiment and incision healed well. The gross observation showed that joint synovia increased and articular surface was smooth and integrity in group A; that ulcer was observed on the articular surface of group B; and that articular surface was smooth and integrity in group C. There were sigificant differences in articular cartilage scores between 3 groups (P lt; 0.05). The histological observation showed that the articular cartilage had normal structure and the cells arranged regularly in group A; that the articular cartilage became thin and the cells arranged irregularly in group B; and that the cells arranged with a clear layer and had regular shape in group C. The Mankin scores were 1.04 ± 0.13, 7.97 ± 0.12, and 2.81 ± 0.36 in groups A, B, and C, respectively; showing significant difference between 3 groups (P lt; 0.05). The result of immunohistochemistry showed that the expressions of TGF-β1 were 50.62 ± 1.51, 24.81 ± 1.28, and 41.57 ± 1.69 and the expressions of IL-1β were 13.12 ± 1.21, 62.53 ±2.37, and 30.67 ± 1.28; showing significant differences between 3 groups (P lt; 0.05). Conclusion A daily administration ofOTL at a dose of 150 mg/kg for 12 weeks can partially decrease the expression levels of IL-1β and increase the expression levels of TGF-β1, which delays the development of osteoarthritis.
Objective To observe the effect of Melittin on collagen type II (Col-II) expression of rat endplate chondrocytes (EPCs) induced by interleukin 1β (IL-1β). Methods Primary EPCs from the lumbar vertebra of 4-week-old Sprague Dawley rats were culturedin vitro and identified by morphological observation, toluidine blue staining and Col-II immunofluorescence staining. Then, MTT assay was used to determine the optimal concentration of IL-1 and Melittin. Next, EPCs at passage 3 were randomly divided into 4 groups: no treatment was done in group A as control group; the optimal concentration of IL-1β, Melittin, and both IL-1β and Melittin were used in groups B, C, and D respectively. The expression of Col-II was detected by Western blot after 48 hours intervention. Results Under inverted microscope, the first generation EPCs were polygonal; cell proliferation decreased after fifth generation, and cell morphology changed into fusiform. The acidic mucosubstance in the cytoplasm (such as Aggrecan) was stained dark blue by toluidine blue. After marking Col-II by immunofluorescence, the positive expression of cytoskeleton (green fluorescence) could be observed. MTT assay showed that IL-1β and Melittin could inhibit the EPCs in a dose-dependent manner after intervention of 24 and 48 hours, and the optimal concentrations of IL-1β and Melittin intervention were 10 ng/mL and 1.0 μg/mL respectively. Compared with group A, the expression of Col-II was significantly reduced in group B, and was significantly increased in group C by Western blot assay, but there was no significant difference between group D and group A. The Col-II expression levels of groups A, B, C, and D were 0.991±0.024, 0.474±0.127, 1.913±0.350, and 1.159±0.297 respectively, showing significant difference between the other groups (P<0.05) except between group A and group D (P>0.05). Conclusion Melittin has a protective effect on endplate cartilage, and the research results provide experimental basis for the prevention and treatment of spinal degenerative disease.
Objective To investigate the effect and mechanism of miR-4287, a chondrogenesis associated microRNA, regulated the expression of aggrecanase-1 (a disintegrin and metalloproteinase with thrombospondin motif 4, ADAMTS4) in human chondrocytes. Methods First, the voluntarily donated normal and osteoarthritic knee articular cartilages were used to detect the expressions of miR-4287 and ADAMTS4 mRNA by real-time fluorescence quantitative PCR. Then, chondrocytes were isolated from knee articular cartilages. The effect of IL-1β on the expression of miR-4287 and ADAMTS4 mRNA was validated by the first generation of osteoarthritic chondrocytes. To confirm the influence of IL-1β signal pathways on the expression of miR-4287 and ADAMTS4 mRNA, osteoarthritic chondrocytes were pretreated with MAPK signal pathway inhibitor SP600125, NF-κB pathway inhibitor SN50, and finally stimulated with IL-1β. Chondro cytes were transfected with miR-4287 mimics and mimics negative control, inhibitors and inhibitors negative control respectively to value the effect of miR-4287 on ADAMTS4 expression. Luciferase reporter assay was used to verify the direct interaction between miR-4287 and putative site in the 3-untranslated region (3’UTR) of ADAMTS4 mRNA. Results Compared with normal knee articular cartilages, the miR-4287 expression was markedly diminished and conversely ADAMTS4 mRNA expression was raised in osteoarthritis cartilages (P<0.05). Stimulation with IL-1β led to a reduction in miR-4287 expression and upregulation in ADAMTS4 mRNA expression, showing significant difference when compared with the untreated groups (P<0.05). Pretreatment with IL-1β signal pathway inhibitors induced miR-4287 expression and attenuated ADAMTS4 mRNA expression in human chondrocytes, which were significantly different from that of unstimulated cells (P<0.05). ADAMTS4 mRNA and protein were suppressed by transfection with miR-4287 mimics (P<0.05) and elevated by transfection with miR-4287 inhibitors (P<0.05). As luciferase reporter assay showed, overexpression miR-4287 failed to alter the luciferase activity of a reporter construct containing either wild or mutant 3’UTR of ADAMTS4 mRNA (P>0.05). Conclusion miR-4287, a chondrogenesis associated microRNA, may play an important role in cartilage degeneration. miRNA-4287 is able to regulate ADAMTS4 expression in human chondrocytes, but not by means of directly targeted the ADAMTS4 mRNA 3’UTR. The exact mechanisms need to be further addressed.
Objective To investigate interleukin-1β (IL-1β), IL-6, and IL-17 levels in both synovial fluid and serum of patients with primary knee medial osteoarthritis (OA) after high tbial osteotomy (HTO). Methods Twenty-six patients with primary knee medial OA undergoing HTO between January 2011 and June 2014 (experimental group) and 30 healthy individuals (control group) were recruited into the study. There was no significant difference in gender, age, and body mass index between 2 groups (P>0.05). The X-ray film was taken to record healing time at osteotomy site, to measure the tibiofemoral angle, and to assess limb alignment after HTO. Visual analogue scale (VAS) pain score and knee society score (KSS) were used to evaluate pain level and function of the knee. The IL-1β, IL-6, and IL-17 concentrations in both plasma and synovial fluid were measured before operation and at 6, 12, and 18 months after operation in the experimental group using ELISA method; the levels in plasma were measured in control group. Results Primary healing of incisions was achieved in patients. All patients were followed up 18-24 months (mean, 21 months). The X-ray film showed osseous healing at osteotomy site at 9-14 weeks (mean, 11.5 weeks). The average tibiofemoral angle was 167.5° (range, 165-170°) after bone healing. Satisfactory limb alignment was obtained in all patients. The postoperative VAS pain score was significantly decreased and KSS score was significantly improved when compared with preoperative scores (P<0.05), but no significant difference was found between different time points after operation (P>0.05). The preoperative plasma and synovial fluid IL-1β, IL-6, and IL-17 concentrations were significantly higher in patients than controls (P<0.05). The postoperative IL-1β, IL-6, and IL-17 concentrations in plasma and synovial fluid were significantly lower than preoperative ones in patients (P<0.05), but the concentrations were significantly higher than those in controls (P<0.05). The postoperative plasma and synovial fluid IL-1β, IL-6, and IL-17 concentrations were significantly declined in patients, but there was no significant difference between different time points after operation (P>0.05). Conclusion HTO can significantly improve the pain symptom and joint function and reduce IL-1β, IL-6, and IL-17 levels in both plasma and synovial fluid of patients with medial compartment knee OA, but these cytokines can not return to normal level.