ABSTRACT
Objective To investigate the effects of cisplatin on the expression of nucleotide － binding oligomerization domain－like receptor(NOD) 1 and 2 in human osteosarcoma SaOS－2 cell line,and to explore the mechanism of cisplatin in the treatment of human osteosarcoma. Methods CCK－8 assay,real－time quantitative reverse transcription polymerase chain reaction ( qRT － PCR ) and immumofluorescence methods were used to determine the growth survival rate and expression levels of NOD1 and NOD2 in osteosarcoma SaOS －2 cell line treated with cisplatin (0,5,10,20 μmol/L,named group S0,group S5,group S10,group S20,respectively) for 24, 48,72 hours. Results After treatment with cisplatin for 48 h or 72 h,the growth survival rates of SaOS－2 cells were significantly decreased in group S5 than those in group S0(65. 53% vs. 100. 00%;46. 43% vs. 100. 00%,χ2=8. 64,73. 97,all P＜0. 01). Moreover,after treatment with cisplatin for 24 h,48 h or 72 h,the growth survival rates of SaOS－2 cells were significantly decreased in group S10 or group S20 than those in group S0(80. 60% vs. 100. 00% , 42. 94 vs. 100. 00% ,27. 90% vs. 100. 00% ;62. 54% vs. 100. 00% ,33. 09% vs. 100. 00% ,22. 95% vs. 100. 00% , χ2=20. 99,79. 72,112. 50;45. 40,67. 56,125. 20,all P＜0. 01),and the growth survival rates were significantly lower in group S20 than those in group S5(62. 54% vs. 93. 78% ,33. 09% vs. 65. 53% ,22. 95% vs. 46. 43% ,χ2= 28. 47,21. 78,11. 71,all P ＜0. 01). The expression levels of NOD1 mRNA and NOD2 mRNA in group S5 were significantly increased at 48 h or 72 h than those at 24 h,and were higher than group S0 when treated with 5μmol/L cisplatin[(3. 64 ± 0. 44) vs. (4. 47 ± 1. 22) vs. (1. 79 ± 0. 44) vs (1. 00 ± 0. 00);(6. 88 ± 2. 79) vs. (6. 86 ± 2. 40) vs (2. 29 ± 0. 70) vs. (1. 00 ± 0. 00),F=29. 12,24. 11,all P＜0. 01]. And the expression levels of NOD1 protein had an increased tendency after 48 h or 72 h treatment with 5μmol/L cisplatin. Furthermore,the expression level of NOD1 mRNA was positively correlated with NOD2 mRNA(n=36,r=0. 92,P＜0. 01). Conclusion Cisplatin can elevate the function of osteosarcoma cell in a dose － and time － dependent manners, cisplatin may act as a efficient drug to cure osteosarcoma desease,which may be related to NOD1 and NOD2 signal pathway.
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Objective To investigate the potential roles of nucleotide -binding oligomerization domain (NOD)-like receptor in the pathogenesis of asthma.Methods Through rat asthma model,24 rats were randomly divided into three groups on average,named asthma group,control group and dexamethasone group.Expression levels of NOD1 and NOD2 mRNA were detected by Real -time PCR in lung tissues.Results The expression levels of NOD1 mRNA in the asthma group,control group and dexamethasone group were (0.62 ±0.34)RQ value,(1 .00 ± 0.00)RQ value,(0.65 ±0.33 )RQ value respectively.The levels of NOD1 mRNA in the asthma group was significantly lower than that in the control group(F =4.75,P 0.05).Moreover,expression levels of NOD2 mRNA in the asthma group,control group and dexamethasone group were (0.92 ±0.32)RQ value, (1 .00 ±0.00)RQ value,(1 .50 ±0.56)RQ value,respectively.There was no statistically significant difference of NOD2 mRNA level between the asthma group and control group (P >0.05 ),but level of NOD2 mRNA in the dexamethasone group was significantly higher than that in the asthma group(F =5.64,P 0.05).Conclusion Expression of NOD -like receptor subtype was not at the same level,and their reaction to dexamethasone were different either.
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Objective To investigate the potential roles of pattern recognition receptor in the pathogenesis of asthma,the concentrations of serum nucleotide -binding oligomerization domain like receptors 1 (NOD1 )and Toll like receptor 1(TLR1)and Dectin and sCD14 protein were determined.Methods Blood samples were obtained from 45 cases of severe asthma(asthma group)during acute attack and convalescent period,the concentrations of serum NOD1,TLR1,Dectin and sCD14 protein were assessed using enzyme linked immunosorbent assay(ELISA),and compared with 30 healthy children(control group).Results The concentrations of NOD1,TLR1,Dectin and sCD14 protein were significantly higher in acute attack period of the asthma group[(65.53 ±19.95)ng/mL,(10.46 ±3.35)ng/mL, (80.38 ±19.51)ng/mL,(4.51 ±1.29)ng/mL]than those in the control group[(25.57 ±9.64)ng/mL,(5.54 ± 1.49)ng/mL,(47.37 ±15.16)ng/mL,(2.21 ±0.68)ng/mL](t =11.57,8.63,7.82,9.95,all P 0.05).Furthermore,level of NOD1 was positively corre-lated with sCD14(r =0.49,P <0.01).Conclusion Levels of serum NOD1,TLR1,Dectin and sCD14 protein were increased in acute severe asthma,whereas,they were decreased in convalescent period.The elevation of them indicated that pattern recognition reception has synergistic function in the pathogenesis of asthma attack.
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Objective To investigate the effects of dexamethasone on expressions of epithelial neutrophil activating protein-78 (ENA-78) and transforming growth factor- beta 1 (TGF-β1) in neutrophil of asthma rats.Methods Thirty male SD rats were randomly divided into three groups on average, including asthma group, control group and dexamethasone treated group. In this experiment, the rat model of asthma were established by sentization and challenge with ovalbumin. Blood neutrophil were isolated and purified. The expression of ENA-78 was detected by flow cytometry. The expression of TGF-β1 was detected by immunohistochemical method in blood neutrophil and bronchial wall. Results Expression of ENA-78 in blood neutrophil in dexamethasone treated group(71.82 ±8. 87 mean fluorescence intensity)was lower than that in asthma group, but higher than that in control group(all P ＜0. 01). And expressions of TGF-β1 protein in dexamethasone treated group(0. 173 ± 0. 014,0. 202 ± 0. 019 optical density, respectively) was lower than that in asthma group(all P ＜0. 01) ,but higher than that in control group(all P ＜0. 01). There were significant positive correlation between ENA-78 expression at blood neutrophil and numbers of total inflammation cells in bronchoalveolar lavage fluid (n = 29, γ = 0. 762, P ＜ 0. 01). Conclusion The beneficial effect of glucocorticoid(dexamethasone) on airway inflammation in asthma rats could be at least in part due to their direct inhibitory effect on ENA-78 and TGF-β1 protein generation by neutrophil.
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Objective To observe the expressions of grouth-related oncogen (GRO)α, epithelial neutrophil activating protein-78 (ENA-78) and neutrophil-activating peptide-2 (NAP-2) of rat asthma. And to investigate the role of neutrophil in the pathogenesis of asthma exacerbation. Methods In this experi-ment, the rat model of asthma were randomly divided into two groups on average, including asthma group and control group. Levels of ENA-78 at blood neutrophil were detected by flow cytometry method. The ex-pressions of GROα protein at bronchial wall and NAP-2 protein at blood neutrophil were detected by immuno-histochemieal method. Results Levels of GROα, ENA-78 and NAP-2 proteins in asthma group [0.138 ±0.009(A value), 97.65±13.99(MFI), 0.198±0.016(A value), respectively]were significantly higher than those in control group[0.077±0.010(A value), 50.79±8.66(MFI), 0.079±0.015(A value), re-spectively], all P < 0.01. Conclusion Levels of GROα, ENA-78 and NAP-2 were increased at rat asth-ma. They may be participate in inflammation of asthma exacerbation. Neutrophil may promote inflammatory cells influxing into airway wall via increasing synthesis of CXC chemotactic factors.