| Object:To investigate the role and mechanism of caspase recruiting domain protein 9(CARD9)in the early stage of acute and severe ischemic stroke.Methods:Peripheral venous blood was collected from 36 patients with acute moderate and severe ischemic stroke on day 1,3 and 7.Meanwhile,20 healthy volunteers were selected as the control group and peripheral venous blood was collected.Mononuclear cells were centrifuged by density gradient method.The expression level of CARD9 and the activity of NF-k B in monocytes were determined by Western blot(p-p65/p65).RT-PCR detection of inflammatory factors TNF-a,IL-6,IL-l b mRNA expression;SPSS 23 was used for statistical analysis.Results:Western blot results showed that the expression of CARD9 in peripheral mononucleocytes of patients with ischemic stroke was increased,and on day 1,3,and 7(1.17±0.14,1.08±0.15,and 0.97±0.16)was significantly higher than that of the normal control group(0.34±0.02).In the case group,CARD9 expression reached a peak rapidly on the first day of comparison,and then gradually decreased on the third and seventh days,with significant difference in p-pair comparison(p(27)0.01).Compared with the normal control group(1.21±0.12),the activity of NF-kB on day1,3 and 7(1.78±0.06?1.70±0.07?1.46±0.04)increased after cerebral ischemia.On the 7th day,p(27)0.05 showed a statistical difference.The expression of p-p65 in the patient group reached a peak on the 1st day of the comparison,and then gradually decreased on the 3rd and 7th day of the comparison,with a significant difference in the declining trend in paired comparison(p(27)0.01).The RT-PCR results showed that,compared with the control group(1.00±0.08),TNF-mRNA in the case group(5.34±0.14?4.57±0.15?3.58±0.19)increased on day1,3,and7,showing significant statistical significance(p(27)0.01).Through single factor repetitive measure analysis of variance,1,3,7 day TNF-a mRNA expression is on the decline,compared with the two significant differences(p(27)0.05),IL-1 b mRNA expression(1.00±0.11)compared with control group,1,3,7 days(6.49±0.12?4.85±0.23?3.38±0.31)higher expression,has significant statistical significance(p(27)0.01),and a downward trend over time,the two comparison has significant difference(p(27)0.05);IL-6 mRNA in the case group also showed a decreasing trend with time(p(27)0.05),and significantly increased on day 1,3 and7(10.29±0.55?7.32±0.44?4.56 ± 0.33)compared with the control group(1.00±0.07),showing significant statistical significance(p(27)0.01).Pearson correlation analysis showed that patients with cerebral apoplexy in PBMC of NF-kB activation,the TNF-?,IL-6,IL-lb and CARD9 express mRNA and NIHSS score was positively related to(r = 0.971,0.974,0.970,0.982,r = 0.891,p(27)0.01).Conclusion:1.After ischemic stroke,CARD9 protein expression and NF-k B activity increased,peaked on the first day,and then decreased gradually with time.2.The expression of the inflammatory factors TNF-a,IL-1b and IL-6 increased after ischemic stroke,reached a peak on the first day,and then gradually decreased with time.3.NIHSS score was significantly increased after moderate to severe ischemic stroke,and did not significantly increase or decrease with treatment progress.4.CARD9 was positively correlated with NF-kB and the inflammatory factors TNF-a,IL-l b and IL-6,suggesting that CARD9 activates the NF-k B signaling pathway in the early stage of moderate to severe ischemic stroke to induce the production of inflammatory factors,thereby aggravating brain injury... |
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