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目的观察胸腔内注入尿激酶对结核性胸膜炎的炎症反应的影响。方法用标准人型结核分枝杆菌菌株H37RV0.03 mg/只注入90只Wistar大鼠胸腔内,24 h后随机分成2组。实验组:胸腔内注入尿激酶600 U/(ml.只);对照组:胸内注入生理盐水1 ml/只。在注入后8 h2、4 h3、2 h4、8 h、3 d、5 d、7 d、10 d和15 d分别处死动物,每次每组各5只。解剖胸腔,记录胸腔积液量,观察胸腔、胸膜大体及镜下病理改变。检测胸腔积液中白细胞计数(WBC)及分类、总蛋白(TP)、葡萄糖(GLU)和乳酸脱氢酶(LDH)的含量;测定可溶性细胞间粘附分子-1(sICAM-1)、转化生长因子-β1(TGF-β1)和γ干扰素(IFN-γ)的水平;同时测定外周血的凝血酶原时间(PT)、凝血酶时间(TT)和部分活化凝血酶原时间(APTT);测量胸膜厚度并对胸膜、肺组织作病理学观察。结果实验组第1 d和2 d胸腔积液量[(6.1±0.6)ml和(7.0±0.2)ml]明显多于对照组[(5.4±0.5)mml和(5.2±0.2)ml],第7 d和10d则明显低于对照组[(2.4±0.4)ml和(0.3±0.1)mlVS(5.2±0.5)ml和(3.3±0.6)ml,P<0.01]。实验组胸腔积液的WBC在初期(8 h)和48 h后均明显低于对照组(P<0.01),中性粒细胞百分率也明显低于对照组(P<0.01)。实验组炎症标志物LDH在8 h2、4 h、32 h内明显低于对照组[(15.5±0.7、16.1±1.2、17.5±1.4VS18.0±0.9、18.4±0.6、18.5±1.2)μmol/(s.L),P<0.01]。实验组胸腔积液sICAM-1水平在整个观察期均明显低于对照组(P<0.01),而TGF-β1仅在第5 d和7 d低于对照组[(29.4±3.3、22.7±3.4VS45.6±3.2、45.2±2.7)ng/ml,P<0.01];IFN-γ在第7 d也明显低于对照组[(151.6±21.4VS178.2±18.6)pg/ml,P<0.01]。IFN-γ/TGF-β1在第3 d后明显高于对照组,第7 d最高(6.74±0.9VS3.15±1.8,P<0.01)。同时还观察到实验组胸腔内粘连带的形成时间较迟而且数量也较少,胸膜厚度较薄(P<0.01)。外周血凝血功能与对照组比较无显著性差异。结论胸腔内注入尿激酶可使结核性胸膜炎早期胸液产生较多而后吸收较快,胸腔内炎症反应减轻,从而减轻胸膜粘连和增厚,对凝血功能无影响,可用于结核性胸膜炎的治疗。
Objective To observe the effect of intrapleural injection of urokinase on the inflammatory reaction of tuberculous pleurisy. Methods The standard human Mycobacterium tuberculosis strain H37RV 0.03 mg / was injected into the thoracic cavity of 90 Wistar rats, and then randomly divided into 2 groups 24 hours later. Experimental group: Thoracic injection of urokinase 600 U / (ml. Only); control group: intrathoracic injection of saline 1 ml / only. At 8 h, 4 h, 2 h, 4 h, 8 h, 3 d, 5 d, 7 d, 10 d and 15 d after injection, animals were sacrificed, 5 animals in each group. Dissect the thoracic cavity, record the amount of pleural effusion, and observe the pathological changes of thorax, pleura and microscopic. The levels of WBC and its classification, total protein (TP), glucose (GLU) and lactate dehydrogenase (LDH) in pleural effusion were determined. The levels of sICAM-1, (PT), thrombin time (TT) and partial activated prothrombin time (APTT) in peripheral blood were measured at the same time. The levels of TGF-β1, IFN- ; Measurement of pleural thickness and pleural, lung tissue for pathological observation. Results The volume of pleural effusion on day 1 and day 2 in experimental group was significantly higher than that in control group [(6.1 ± 0.6) ml and (7.0 ± 0.2) ml] [(5.4 ± 0.5) mml and (5.2 ± 0.2) ml] 7d and 10d were significantly lower than that of the control group [(2.4 ± 0.4) ml and (0.3 ± 0.1) mlVS (5.2 ± 0.5) ml and (3.3 ± 0.6) ml, P <0.01]. WBC in the experimental group was significantly lower than that in the control group (P <0.01) and the percentage of neutrophils was significantly lower in the experimental group than that in the control group (P <0.01). LDH in the experimental group was significantly lower than that in the control group within 8 h, 4 h, 32 h [(15.5 ± 0.7, 16.1 ± 1.2, 17.5 ± 1.4VS18.0 ± 0.9, 18.4 ± 0.6, 18.5 ± 1.2) μmol / (sL), P <0.01]. The levels of sICAM-1 in pleural effusion in experimental group were significantly lower than those in control group (P <0.01), while TGF-β1 was only lower on the 5th and 7th day in the experimental group [(29.4 ± 3.3,22.7 ± 3.4 VS45.6 ± 3.2,45.2 ± 2.7) ng / ml, P <0.01]; IFN-γ was also significantly lower than that of the control group [(151.6 ± 21.4 vs 178.2 ± 18.6) pg / ml on day 7, P <0.01 ]. The level of IFN-γ / TGF-β1 in the third day was significantly higher than that in the control group, the highest on the seventh day (6.74 ± 0.9 VS3.15 ± 1.8, P <0.01). At the same time, it was also observed that the formation of intrathoracic adhesions in the experimental group was delayed and less in number, and the pleural thickness was thinner (P <0.01). No significant difference in peripheral blood coagulation compared with the control group. Conclusion Intrapleural injection of urokinase can make tuberculous pleurisy produce more pleural fluid in the early stage and absorb faster, reduce the inflammatory reaction in the thoracic cavity, relieve pleural adhesions and thickening, and have no effect on the coagulation function. It can be used in the treatment of tuberculous pleurisy.