[1]张晓光,党 萍,霍 琳,等.衔接蛋白失能同源物2通过抑制NOD样受体热蛋白结构域相关蛋白3对结核性胸膜炎大鼠炎症和氧化应激的影响[J].陕西医学杂志,2024,(4):468-474.[doi:DOI:10.3969/j.issn.1000-7377.2024.04.007]
　ZHANG Xiaoguang,DANG Ping,HUO Lin,et al.Effects of adaptor protein disabling homologue 2 on inflammation and oxidative stress in rats with tuberculous pleurisy by inhibiting NLRP3[J].,2024,(4):468-474.[doi:DOI:10.3969/j.issn.1000-7377.2024.04.007]
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衔接蛋白失能同源物2通过抑制NOD样受体热蛋白结构域相关蛋白3对结核性胸膜炎大鼠炎症和氧化应激的影响
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年4期
页码:
468-474
栏目:
基础研究
出版日期:
2024-04-05

文章信息/Info

Title:
Effects of adaptor protein disabling homologue 2 on inflammation and oxidative stress in rats with tuberculous pleurisy by inhibiting NLRP3
作者:
张晓光1; 党 萍2; 霍 琳3; 刘 会4
(1.河北省胸科医院内科,河北 石家庄 050041; 2.河北省胸科医院结核内科,河北 石家庄 050041; 3.河北省胸科医院药学部,河北 石家庄 050041; 4.河北省胸科医院感染性疾病科,河北 石家庄 050041)
Author(s):
ZHANG Xiaoguang; DANG Ping; HUO Lin; LIU Hui
(Department of Internal Medicine,Hebei Chest Hospital,Shijiazhuang 050041,China)
关键词:
结核性胸膜炎;  肺外结核病;  衔接蛋白失能同源物2;  NOD样受体热蛋白结构域相关蛋白3;  炎症;  氧化应激
Keywords:
Tuberculous pleurisy;  Extrapulmonary tuberculosis;  Adaptor protein disabling homologue 2;  Nod-like receptor pyrin domain-containing protein 3;  Inflammation;  Oxidative stress
分类号:
R -332
DOI:
DOI:10.3969/j.issn.1000-7377.2024.04.007
文献标志码:
A
摘要:
目的:探讨衔接蛋白失能同源物2(DAB2)抑制NOD样受体热蛋白结构域相关蛋白3(NLRP3)对结核性胸膜炎大鼠炎症和氧化应激的影响。方法:按照随机数字法将60只SPF 级雄性SD大鼠分为四组,每组40只。除正常对照组外,结核性胸膜炎组、DAB2组和pcDNA- NLRP3组进行建模处理,以第2天是否抽出胸腔积液为模型建立成功。正常对照组不注射结核分枝杆菌 H37RV悬液,DAB2组建模后第2天静脉注射AAV9-DAB2质粒,每天1次,连续注射7 d,DAB2+pcDNA-NLRP3组在注射AAV9-DAB2质粒500 μg/L的同时注射pcDNA-NLRP3 80 μl,正常对照组和结核性胸膜炎组的大鼠尾静脉注射0.9%氯化钠溶液。进行各组呼吸功能指标测定,收集胸腔积液,记录积液量,观察3、5、7 d的胸腔积液粘连性情况,HE染色观察胸膜组织病理学情况,Western blot检测胸膜组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、基质金属蛋白酶1(MMP-1)和MMP-9蛋白表达,荧光探针DCFH-DA分析检测活性氧(ROS)的水平。结果: 与正常组相比,结核性胸膜炎组的大鼠的胸腔积液和胸膜厚度明显增加,用力肺活量(FVC)、最大呼气流量(PEF)、用力呼气容积(FEV)0.3和FEV0.3/FVC显著降低,TNF-α、IL-8、MMP-1和MMP-9蛋白表达显著升高,基质金属蛋白酶抑制剂(TIMP-1)蛋白表达显著降低,丙二醛(MDA)水平升高,超氧化物歧化酶(SOD)水平降低,ROS累积量明显升高(均P<0.001); 与结核性胸膜炎组相比,DAB2组大鼠胸腔积液和胸膜厚度显著降低,FVC、PEF、FEV0.3和FEV0.3/FVC明显升高,DAB2组大鼠胸膜组织中的TNF-α、IL-8、MMP-1和MMP-9蛋白表达明显降低,TIMP-1水平明显升高,MDA水平降低,SOD水平升高,ROS累积量明显降低(均P<0.001); 与DAB2组相比,DAB2+pcDNA- NLRP3组大鼠胸腔积液和胸膜厚度明显升高,FVC、PEF、FEV0.3和FEV0.3/FVC明显降低,DAB2+pcDNA-NLRP3组的TNF-α、IL-8、MMP-1和MMP-9蛋白表达明显升高,TIMP-1蛋白表达显著降低,MDA水平明显升高,SOD水平显著降低,ROS累积量显著升高(均P<0.001),各组大鼠在3、5、7 d的胸腔积液粘连性评分比较采用重复测量设计的方差分析,结果显示,不同时间点的胸腔积液粘连性评分比较差异具有统计学意义(均P<0.001); 各组胸腔积液粘连性评分比较差异具有统计学意义(均P<0.001); 各组胸腔积液粘连性评分变化趋势比较差异有统计学意义(均P<0.001); 与模型组相比,DAB2组大鼠的胸膜内和肺间质内血管充血症状减轻,有少量的纤维组织增生,上皮样细胞团以及凝固型坏死也明显减少,淋巴细胞和中性粒细胞浸润也明显减少; 与DAB2组相比,DAB2+pcDNA-NLRP3组大鼠的胸膜内和肺间质内血管充血症状加重,出现的纤维组织增生,上皮样细胞团以及凝固型坏死也明显增多,淋巴细胞和中性粒细胞浸润也明显增加。结论: DAB2通过抑制NLRP3的活性促进胸腔积液的吸收,降低胸膜厚度和粘连发生率,降低炎症反应和氧化应激水平,缓解结核性胸膜炎的进展。
Abstract:
Objective:To investigate the effect of adaptor protein disabling homologue 2(DAB2)inhibiting NLRP3 on inflammation and oxidative stress in rats with tuberculous pleurisy.Methods:60 SPF male SD rats were divided into 4 groups and DAB2+pcDNA-NLRP3 group with 40 rats per group.In addition to the normal control group,the tuberculous pleurisy group,DAB2 group and pcDNA-NLRP3 group were modeled,and the model was successfully established based on whether pleural effusion was extracted on the 2nd day.The normal control group was not injected with mycobacterium tuberculosis H37RV suspension,and the AAV9-DAB2 plasmid was injected intravenously on the second day after the formation of the model,once a day for 7 days.The DAB2+pcDNA-NLRP3 group was injected with AAV9-DAB2 plasmid 500 μg/L and pcDNA-NLRP3 80 μl at the same time,and the rats in the normal control group and the tuberculous pleuritis group were injected with 0.9% sodium chloride solution through the tail vein.Respiratory function indexes of each group were measured,pleural effusion was collected,the amount of effusion was recorded,and the adhesion of pleural effusion was observed at 3,5 and 7 days.The pleural histopathology was observed by HE staining,and the protein expressions of TNF-α,IL-8,MMP-1 and MMP-9 in pleural tissue were detected by Western blot.Fluorescence probe DCFH-DA was used to detect ROS level.Results:Compared with the normal group,the pleural effusion and pleural thickness of the rats in the tuberculous pleuritis group were significantly increased,FVC,PEF,FEV0.3 and FEV0.3/FVC were significantly decreased,the protein expressions of TNF-α,IL-8,MMP-1 and MMP-9 were significantly increased,the protein expression of TIMP-1 was significantly decreased,and the level of MDA was increased.SOD level decreased,ROS accumulation increased significantly(all P<0.001).Compared with the tuberculous pleuritis group,pleural effusion and pleural thickness of DAB2 group were significantly decreased,FVC,PEF,FEV0.3 and FEV0.3/FVC were significantly increased,the protein expressions of TNF-α,IL-8,MMP-1 and MMP-9 in pleural tissue of DAB2 group were significantly decreased,and the level of TIMP-1 was significantly increased.MDA level decreased,SOD level increased,ROS accumulation decreased significantly(all P<0.001).Compared with DAB2 group,pleural effusion and pleural thickness in DAB2+pcDNA-NLRP3 group were significantly increased,while FVC,PEF,FEV0.3 and FEV0.3/FVC were significantly decreased.In DAB2+pcDNA-NLRP3 group,the expressions of TNF-α,IL-8,MMP-1 and MMP-9 proteins were significantly increased,the expression of TIMP-1 protein was significantly decreased,the level of MDA was significantly increased,the level of SOD was significantly decreased,and the accumulation of ROS was significantly increased(all P<0.001).Repeated measure design analysis of variance was used to compare the scores of pleural effusion at 3,5 and 7 days.The results showed that there were statistically significant differences in the scores of pleural effusion at different time points(all P<0.001).There were statistically significant differences in the score of adhesion of pleural effusion among all groups(all P<0.001).There was statistical significance in the score of pleural effusion adhesion in all groups(all P<0.001).Compared with the model group,the symptoms of intrapleural and interstitial vasocongestion in DAB2 group were reduced,there was a small amount of fibrous tissue hyperplasia,epithelioid cell mass and coagulation necrosis were also significantly reduced,and lymphocyte and neutrophil infiltration were also significantly reduced.Compared with DAB2 group,intrapleural and interstitial pulmonary vasocongestion in DAB2+pcDNA-NLRP3 group was aggravated,fibrous tissue hyperplasia,epithelioid cell mass and coagulation necrosis were also significantly increased,and lymphocyte and neutrophil infiltration were also significantly increased.Conclusion: DAB2 can promote the absorption of Pleural effusion by inhibiting the activity of NLRP3,reduce the thickness of pleura and the incidence of adhesion,reduce inflammatory reaction and oxidative emergency level,and alleviate the progress of tuberculous pleurisy.

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备注/Memo

备注/Memo:
基金项目:河北省医学科学研究重点计划课题(20160503)
更新日期/Last Update: 2024-04-07