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目的观察乌司他丁对大鼠早期放射性肺损伤的防治作用及其机制。方法选取45只健康雌性SD大鼠,根据随机数字表法将大鼠分为正常组、模型组和实验组,每组15只。正常组不做处理,其余2组均全胸接受6 MV-X射线单次照射20 Gy制作放射性肺损伤模型。实验组大鼠予照射后48 h起予以尾静脉注射乌司他丁1×105U·kg-1qd,其余2组予以尾静脉注射同等体积0.9%Na Cl溶液。2周后留取肺组织,苏木精-伊红(HE)染色观察肺组织形态,计数肺泡灌洗液白细胞和红细胞数,实时荧光定量聚合酶链式反应(qRT-PCR)和免疫印迹法(Western blot)测定高迁移率族蛋白B1(HMGB1)、晚期糖基化终末产物受体(RAGE)、核转录因子kappa B(NF-κB)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和环氧合酶2(COX-2)的表达水平。结果 HE染色结果显示,实验组大鼠肺泡炎症程度较模型组显著减轻,实验组肺泡灌洗液中白细胞和红细胞数分别(3.42±1.53)×109/L,(3.95±1.24)×109/L,模型组为(7.04±2.18)×109/L,(7.12±1.35)×109/L,实验组均较模型组明显减少(P<0.05),但与正常组细胞数无明显差异(P>0.05)。正常组大鼠肺组织中HMGB1、RAGE、NF-ΚB、TNF-α、IL-6和COX-2 mRNA的表达分别为1.01±0.16,1.01±0.18,1.03±0.27,1.10±0.50,1.01±0.17和1.01±0.16;模型组中上述基因mRNA的表达分别为4.66±1.43,4.18±0.82,4.31±1.03,3.41±0.91,4.22±0.79和4.41±0.99;实验组中上述基因mRNA的表达分别为1.82±0.73,1.55±0.62,1.70±1.39,1.77±0.95,1.45±0.55和1.85±1.48。与模型组比较,实验组肺组织HMGB1信号通路表达水平均下降,差异均有统计学意义(均P<0.05);实验组与正常组比较,各基因的表达水平差异无统计学意义(P>0.05)。结论乌司他丁可通过干扰HMGB1信号通路的表达抑制胸部照射后大鼠肺组织炎性改变。
Objective To observe the preventive and therapeutic effects of ulinastatin on early radiation-induced lung injury in rats and its mechanism. Methods Forty-five healthy female Sprague-Dawley rats were randomly divided into normal group, model group and experimental group according to random number table method, with 15 rats in each group. The normal group did not deal with the other two groups were received 6 MV-X-ray full-ray exposure of 20 Gy to make a model of radiation-induced lung injury. Rats in the experimental group were injected ulinastatin 1 × 105U · kg-1qd into the caudal vein 48 hours after irradiation, while the other two groups were given the same volume of 0.9% NaCl solution through the caudal vein. Lung tissue was collected 2 weeks later, the morphology of lung was observed by hematoxylin-eosin (HE) staining, the numbers of white blood cells and erythrocytes in bronchoalveolar lavage fluid were counted, real-time quantitative polymerase chain reaction (qRT-PCR) and western blot The expressions of HMGB1, RAGE, NF-κB, TNF-α, leukocyte Interleukin 6 (IL-6) and cyclooxygenase 2 (COX-2) expression levels. Results The results of HE staining showed that the degree of alveolar inflammation in the experimental group was significantly reduced compared with the model group. The numbers of leukocytes and erythrocytes in the alveolar lavage fluid in the experimental group were (3.42 ± 1.53) × 109 / L and (3.95 ± 1.24) × 109 / L (7.04 ± 2.18) × 109 / L and (7.12 ± 1.35) × 109 / L respectively in the model group compared with the model group (P <0.05), but there was no significant difference between the experimental group and the normal group (P> 0.05). The expressions of HMGB1, RAGE, NF-κB, TNF-α, IL-6 and COX-2 mRNA in the lungs of normal rats were 1.01 ± 0.16, 1.01 ± 0.18, 1.03 ± 0.27, 1.10 ± 0.50 and 1.01 ± 0.17 And 1.01 ± 0.16, respectively. The mRNA expression of the above genes in the model group was 4.66 ± 1.43, 4.18 ± 0.82, 4.31 ± 1.03, 3.41 ± 0.91, 4.22 ± 0.79 and 4.41 ± 0.99, respectively. The mRNA expression of the above genes in the experimental group was 1.82 ± 0.73, 1.55 ± 0.62, 1.70 ± 1.39, 1.77 ± 0.95, 1.45 ± 0.55 and 1.85 ± 1.48. Compared with the model group, the expression level of HMGB1 in the lung tissue of the experimental group decreased significantly (all P <0.05), but there was no significant difference between the experimental group and the normal group (P> 0.05). Conclusion Ulinastatin can inhibit the inflammatory changes of lung tissue in rats after thoracic radiation by interfering the expression of HMGB1 signaling pathway.