血必净注射液对急性百草枯中毒大鼠肺组织中基质金属蛋白酶及其组织抑制因子的影响

Effect of Xuebijing injection on matrix metalloproteinases and its tissue inhibitory factor in lung tissue of rats with acute paraquat poisoning

目的通过观察血必净注射液对急性百草枯中毒大鼠肺组织中超氧化物歧化酶(SOD)、丙二醛(MDA)及基质金属蛋白酶(MMP)-2、MMP-9及其组织抑制因子1(TIMP-1)表达的影响,探索血必净注射液治疗急性百草枯中毒肺损伤的机制。方法将126只Wister大鼠采用随机数字表法分成正常组、染毒组、血必净组,每组42只。染毒组、血必净组均腹腔内注射百草枯进行造模,血必净组造模后每天给予血必净注射液4mL/kg腹腔内注射。分别于实验的第1,3,7,14,21,28,35天,每组各处死6只大鼠,留取肺组织标本,镜下观察病理变化,采用黄嘌呤氧化酶法测定SOD活力、硫代巴比妥酸法检测MDA含量,免疫组化法检测肺组织中MMP-2、MMP-9、TIMP-1蛋白表达水平。结果染毒组肺泡充血水肿,肺间质增厚,炎性细胞大量浸润,第35天时成纤维细胞大量融合,形成纤维化;血必净组肺泡水肿、炎细胞浸润以及肺间质纤维化均好转,病理改变介于染毒组与正常组之间。染毒组大鼠肺组织中SOD活力下降,MDA含量升高,MMP-2、MMP-9、TIMP-1蛋白表达水平第1天即开始上升,MMP-2第7天达到高峰,MMP-9第3天达到高峰,TIMP-1第21天达到高峰,达到高峰后,MMP-2、MMP-9逐渐下降,TIMP-1却维持在高水平,各时段与正常组比较差异均有统计学意义(P均<0. 05)。与染毒组比较,血必净组大鼠肺组织中SOD活力明显升高(P <0. 05),MDA含量及MMP-2、MMP-9、TIMP-1蛋白表达量明显降低(P均<0.05);与正常组比较,SOD活力仍较低(P <0. 05),MDA含量和MMP-2、MMP-9、TIMP-1蛋白表达量仍较高(P均<0. 05)。结论 MMP-2、MMP-9及TIMP-1在百草枯所致大鼠肺损伤中起重要作用,血必净注射液能抑制肺组织中氧自由基反应,降低MMP-2、MMP-9、TIMP-1的表达,减轻染毒大鼠的肺损伤。

Objective It is to observe the effects of Xuebijing injection on the expression of superoxide dismutase(SOD),malondialdehyde(MDA),matrix metalloproteinase(MMP)-2 and MMP-9 its tissue inhibitory factor 1(TIMP-1)in lung tissues of rats with acute paraquat poisoning,thus to explore the mechanism of Xuebijing injection in the treatment of acute paraquat poisoning lung injury.Methods A total of 126 Wister rats were randomly divided into normal group,poisoning group and Xuebijing group,with 42 rats in each group.Both the poisoning group and the Xuebijing group were intraperitoneally injected with paraquat for modeling,and the Xuebijing group was given intraperitoneal injection of Xuebijing injection 4 mL/kg every day after modeling.On the 1st,3rd,7th,14th,21st,28th and 35th day of the experiment,6 rats were killed in each group.The lung tissue samples were taken and the pathological changes were observed under microscope.The SOD activity was determined by xanthine oxidase method.MDA content was detected by thiobarbituric acid method,and the expression levels of MMP-2,MMP-9 and TIMP-1 in lung tissue were detected by immunohistochemistry.Results In the poisoning group,the alveolar congestion and edema,pulmonary interstitial thickening,and infiltration of abundant inflammatory cells were found,On the 35th day,fibroblasts were highly fused to form fibrosis;Xuebijing group had improved alveolar edema,inflammatory cell infiltration and pulmonary interstitial fibrosis,its pathological changes between the poisoning group and the normal group.The activity of SOD decreased and the content of MDA increased in the lung tissue of rats in the poisoning group,the expression levels of MMP-2,MMP-9 and TIMP-1 began to increase on the first day,MMP-2 reached the peak on the 7th day,MMP-9 on the third day,TIMP-1 on the 21st day.After reaching the peak,MMP-2 and MMP-9 gradually decreased,and TIMP-1 remained at a high level.The differences in these indexes were significant between poisoning group and normal group(P<0.05).Compared with the poisoning group,higher activity of SOD,and lower MDA content and the expression of MMP-2,MMP-9 and TIMP-1 protein in the lung tissue was found in Xuebijing group(P<0.05),but compared with the normal group,the SOD activity was still lower(P<0.05),and the MDA content and MMP-2,MMP-9,TIMP-1 protein expression levels were still higher(P<0.05).Conclusion MMP-2,MMP-9 and TIMP-1 play an important role in lung injury induced by paraquat.Xuebijing injection can inhibit the oxygen free radical reaction in lung tissue and reduce the expression of MMP-2,MMP-9 and TIMP-1,thus to relieve lung injury in the poisoning rats.