Objective To observe the human mononuclear cell releasing TNF-α and the activation of Caspase-3 during apoptosis after stimulated by Co2+ and Cr3+, to discuss the mechanism of artificial joint wear production metal ion on aseptic loosening. Methods CoCl2 powder and CrCl3 powder were dissolved by asepsis inject water, preparing solution for10 mg/L and 500 mg/L, respectively. Mononuclear cells were acquired from peripheral blood, 4 × 106/culture dish. According to the difference of culture solution, the cells were divided into 3 groups. Group A: mononuclear cell was culture with normal sal ine as control; group B: mononuclear cell was cultured with CoCl2 solution; group C: mononuclear cell was cultured with CrCl3 solution. The production of TNF-α was assessed by ELISA, the activation of Caspase-3 was measured by colorimetric assay and the apoptotic cell was detected by TUNEL assays at 12, 24 and 48 hours after co-cultured respectively. Results The concentration of TNF-α and the expression of Caspase-3 in groups B and C were higher than those in group A (P lt; 0.05), and reached the peak level at 24, 48 hours, respetively. The TUNEL positive cells were detected in the all groups, nucleus was pyknotic and darker-staining, cell body was crinkle and cell membrane was integrity. There were significant differences in the apoptosis rate between groups B, C and group A (P lt; 0.05). And the activation of Caspase-3 increased and had the positive correlation with the apoptosis rate (r=0.765). Conclusion Co2+ and Cr3+ ions can stimulate human mononuclear cell to release TNF-α and induce human mononuclear cell apoptosis, which result in periprosethetic osteolysis and related to activation of Caspase-3.
ObjectiveTo investigate the expression of tumor necrosis factor α(TNF-α ) in isolated rat heart at different time points after myocardial hypoxia/reoxygenation. MethodsThe isolated langendorff perfused rat heart model was established. Forty-eight SD rats were randomly divided into four groups: a sham group, hypoxia/reoxygenation groups including a H/R 0.5 h group, a 1 h group and a 2 h group. The heart rate(HR), the 1eft ventricular development pressure(LVDP), maximal rates of increase/decrease of the left ventricular pressure(±dp/dtmax) were continuously recorded. The concentrations of TNF-α and creatine kinase-MB(CK-MB) in myocardium, mRNA expression of TNF-α in myocardium were tested. Ultra structure of myocardium was observed under electron microscope. ResultsThe levels of LVDP, ±dp/dtmax, and HR of hypoxia/reoxygenation group were significantly lower than those in the sham group(P<0.05).The levels of TNF-α and CK-MB and the expressions of TNF-α at mRNA level in the hypoxia/reoxygenation group were higher than those in the sham group(P<0.05).There were significant differences in the above parameters among the H/R 0.5 h group, the 1 h group, the 2 h group(P<0.05).The concentrations of TNF-α and CK-MB, the mRNA expression of TNF-α were higher in the I/R 2 h group than those in the other two groups. ConclusionThe high expression of TNF-α in myocardium after myocardial hypoxia/reoxygenation in rats is related to the degree of myocardium damage and may lead to myocardial injury.
Objective To summarize results of the correlation of tumor necrosis factor-α (TNF-α) promoter –308A/G polymorphism with systemic lupus erythematosus (SLE) susceptibility in Chinese populations. Methods We collected all the publications about the correlation between TNF-α promoter –308A/G polymorphism and SLE in Chinese populations by searching PubMed, EBSCO, CBM, CNKI and Wanfang Data before the date of March 20, 2010. Meta-analysis was performed for checking the difference between two groups about genotypes such as AA versus GG, GA versus GG, AA versus GG+GA, GA+AA versus GG, and A allele versus G allele. Results A total of 8 studies involving 731 SLE patients and 901 healthy people were included. The meta-analysis of total populations showed that, there was no significant correlation between A allele and increased SLE risk (OR=1.42, 95%CI 0.97 to 2.09, P=0.07); the meta-analyses of populations in different regions showed there was no significant correlation of A allele and increased SLE risk in Chinese Taiwan populations (OR=1.04, 95%CI 0.77 to 1.40, P=0.82). Moreover, there was no significant difference between SLE group and control group in the genotypes of AA versus GG, GA versus GG, AA versus GG+GA, and GA+AA versus GG.Conclusion This meta-analysis dosen’t demonstrate the correlation between TNF-α promoter–308A/G polymorphism and SLE in Chinese populations.
ObjectiveTo investigate the association between TNF-α gene -308G/A polymorphism and the risk of coronary atherosclerotic heart disease (CHD) in Chinese population.MethodsWe searched PubMed, Web of Science, CNKI, WanFang Data and VIP Databases from inception to February 2017, to collect case-control studies about the association between TNF-α gene -308G/A polymorphism and risk of CHD in Chinese population. Two reviewers independently screened literature, extracted data and assessed the risk of bias of the included studies. Meta-analysis was performed by Stata 12.0 software.ResultsA total of 10 case-control studies were included. The results of meta-analysis showed a significant association between the TNF-α gene -308G/A polymorphism and CHD risk in Chinese population (A vs. G: OR=1.13, 95%CI 1.02 to 1.26, P=0.020; AA vs. GA/GG: OR=1.47, 95%CI 1.02 to 2.12, P=0.038; AA vs. GG: OR=1.50, 95%CI 1.04 to 2.16, P=0.029).ConclusionThe current evidence shows that the TNF-α gene -308G/A polymorphism may be associated with CHD risk in Chinese population and A allele may be a risk factor. Due to the limited quantity and quality of included studies, more high quality studies are needed to verify the above conclusion.
Objective To comprehensively evaluate the association between TNF-α gene ?308 G/A polymorphism and the risk of prostate cancer. Methods A meta-analysis was performed to analyze the association between ?308 G/A polymorphism and the risk of prostate cancer risk. Results A total of 11 case-control studies (4 919 cases and 5 210 controls) were included in this meta-analysis. The result showed no statistically significant differences in all genotype distribution between prostate cancer cases and controls: dominant model (OR=1.11, 95%CI 0.90 to 1.36, P=0.33), recessive model (OR=0.91, 95%CI 0.70 to 1.18, P=0.47), GA versus GG (OR=1.11, 95%CI 0.90 to 1.37, P=0.33), AA versus GG (OR=0.92, 95%CI 0.71 to 1.20, P=0.55), A versus G (OR=1.07, 95%CI 0.91 to 1.26, P=0.39). In the subgroup analysis by ethnicity, no statistically differences were found between prostate cancer cases and controls. Conclusion This results of meta-analysis suggests that TNF-α gene –308G/A polymorphism may not be a risk factor of prostate cancer. Due to the limited quantity of the includied studies, further studies are needed to validate the above conclusion.
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 explore the effect of Vitamin C (Vit C) on the apoptosis of human nucleus pulposus (NP) cells induced by tumor necrosis factor α (TNF-α) and serum deprivation. MethodsThe NP cells were isolated from patients undergoing spine corrective operation by collagenase trypsin. The experiment was divided into 3 groups:Vit C group (group A), TNF-α group (group B), and serum deprivation group (group C). Group A was reassigned to A1 subgroup (basic medium), A2 subgroup (100 μg/mL Vit C), and A3 subgroup (200 μg/mL Vit C). Group B was reassigned to B0 subgroup (control group), B1 subgroup (100 ng/mL TNF-α), B2 subgroup (100 μg/mL Vit C+100 ng/mL TNF-α), and B3 subgroup (200 μg/mL Vit C+100 ng/mL TNF-α). Group C was reassigned to C0 subgroup (Control group), C1 subgroup (2% FBS), C2 subgroup (2%FBS+100 μg/mL Vit C), and C3 subgroup (2% FBS+200 μg/mL Vit C). After C1 subgroup (2% FBS), C2 subgroup (2%FBS+100 μg/mL Vit C), and C3 subgroup (2% FBS+200 μg/mL Vit C). After application of 100 μg/mL or 200 μg/mL Vit C for 24 hours, NP cells were stimulated by TNF-α and serum deprivation, then the apoptosis rate of NP cells was detected by a flow cytometry, and the gene expressions of the extracellular matrix of NP cells (collagen type Ⅰ, collagen type Ⅱ, aggrecan, and Sox9) and apoptosis related genes (p53, FAS, and Caspase 3) were detected by real-time fluoroscent quantitative PCR. ResultsGroup A:Vit C could significantly reduce the apoptosis rate and gene expressions of p53, FAS, and Caspase 3 of NP cells in A2 and A3 subgroups when compared with A1 subgroup (P<0.05), but there was no significant difference between A2 subgroup and A3 subgroup (P>0.05); Vit C could promote the expressions of the extracellular matrix (collagen type Ⅰ, collagen type Ⅱ, aggrecan, and Sox9) of NP cells in a concentration dependent manner (P<0.05). Group B:TNF-α significantly increased the apoptosis rate and the gene expressions of p53, FAS, and Caspase 3 in B1 subgroup when compared with B0 subgroup (P<0.05); however, Vit C significantly increased the apoptosis rate and the gene expressions in B2 subgroup, and significantly decreased them in B3 subgroup when compared with B1 subgroup (P<0.05). Group C:2% FBS significantly increased the apoptosis rate of NP cells and significantly reduced the gene expressions of p53, FAS, and Caspase 3 in C1 subgroup when compared with C0 subgroup (P<0.05); Vit C could significantly reduce the apoptosis rate and gene expressions of p53, FAS, and Caspase 3 in C3 subgroup, but it could significantly increase them in C2 subgroup when compared with C1 subgroup (P<0.05). ConclusionVit C can promote the synthesis and secretion of extracellular matrix of NP cells. 200 μg/mL Vit C may delay the apoptosis induced by TNF-α and serum deprivation, indicating the potential therapeutic effect of Vit C on intervertebral disc degeneration.
ObjectiveTo investigate the role of Wnt5a in the mechanism of radiculopathy and the relation between Wnt5a and tumor necrosis factorα(TNF-α) by observing the change of the expression of Wnt5a in the rat model of chronic compression of dorsal root ganglia (CCD). MethodsA total of 192 adult male Sprague Dawley rats were allocated into 4 groups: shame group (group A, n=48), CCD group (group B, n=48), CCD+sal ine group (group C, n=48), and CCD+etanercept group (group D, n=48). An L-shaped needle (about 3.5 mm in length, 0.6 mm in diameter) was inserted into the L5 intervertebral foramen, and the dorsal root ganglia (DRG) was compressed by the needle to prepare the CCD model in groups B, C, and D, and then normal sal ine (5.5 mg/kg) or etanercept was injected intraperitoneally in groups C and D. The intervertebral foramen was exposed in group A. The mechanical pain threshold of the posterior paw was tested by the von Frey filaments at 1, 3, 5, and 7 days after operation; the expressions of Wnt5a protein and mRNA were detected at 3 and 7 days after operation by immunohistochemical staining and RT-PCR, respectively. ResultsThe mechanical pain threshold of groups B and C was significantly lower than that of groups A and D, and in group D than in group A (P < 0.05), but no significant difference was found between groups B and C (P > 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a at 7 days were significantly more than those at 3 days in groups B, C, and D (P < 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a in groups B and C were significantly more than in groups A and D, and in group D than in group A (P < 0.05), but no significant difference was shown between groups B and C (P > 0.05). ConclusionThe expression of Wnt5a in the DRG is increased after CCD. The expression of Wnt5a in the DRG is decreased after the administration of the inhibitor of TNF-α.
急性肺損傷(acute lung injury,ALI)是臨床常見的呼吸系統急危重癥,其發病機制復雜,病死率高。腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)是ALI發生、發展過程中非常重要的細胞因子。本文就TNF-α在ALI中的作用和以抗TNF-α為靶點治療ALI的相關研究進行綜述。
目的:研究丹參酮ⅡA(Tan ⅡA)對急性早幼粒細胞白血病(APL)細胞株NB4細胞誘導的血管內皮細胞株(ECV304)促凝活性(PCA)的影響,并對其機制作初步探討。方法:(1)分別用1.0μg/mL TanⅡA、0.3μg/mLATRA、0.01%DMSO、PRMI1640處理NB4細胞24、48和72h,取其上清液作為條件培養基(hNB4-CM)。將這些CM分別與ECV304細胞在37oC共同孵育0、4、8和12h,用反復凍融法制備ECV304細胞裂解液,采用一期凝血法測定其PCA;采用ELISA法測定條件培養基中的TNF-α 。(2)ECV304細胞與1.0μg/mL TanⅡA及TanⅡA 72h-NB4-CM 在37oC共同分別孵育6、12、24和48h,并以ATRA和DMSO分別作為陽性和陰性對照,用上述相同方法測定ECV304細胞裂解液的PCA。結果:(1)1.0 μg/mL Tan ⅡA可以誘導NB4細胞分化,其作用NB4細胞的培養基有一定的升高ECV304細胞PCA的作用,該作用在孵育4h時達高峰,之后ECV304細胞PCA逐漸下降。與0.3μg/mL ATRA的作用無統計學差異(Pgt;0.05)。(2)1.0 μg/mL的TanⅡA對TanⅡA72h-NB4-CM促ECV304細胞PCA有抑制作用,其強度隨作用時間增加而增加,與1.0μmol/L ATRA比較,Pgt;0.05。(3)TanⅡA作用NB4細胞的培養基中TNF-α濃度,在作用前7h內隨作用時間增加而增加,與0.3μg/mL ATRA比較無差異(Pgt;0.05)。結論:Tan ⅡA能誘導NB4細胞分化,后者在分化過程中釋放的TNF-α可能與ECV304細胞PCA活性升高有關;Tan-ⅡA又能抑制Tan-ⅡA-NB4-CM增強ECV304細胞PCA的作用。