文献类型：期刊文献

中文题名：水蛭素对特发性肺间质纤维化大鼠肺组织中纤溶酶原抑制剂-1(PAI-1)mRNA及蛋白表达的干预研究

英文题名：Intervention study of hirudin on the expression of plasminogen inhibitor-1(PAI-1) mRNA and protein in lung tissue of rats with idiopathic pulmonary fibrosis

作者：李红[1];谢海彬[2];刘敏[2];沈明霞[2];王海霞[3];马玉坤[3]

第一作者：李红

机构：[1]深圳市罗湖区中医院;[2]甘肃中医药大学附属医院;[3]甘肃中医药大学

第一机构：深圳市罗湖区中医院

年份：2019

卷号：11

期号：5

起止页码：1

中文期刊名：中医临床研究

外文期刊名：Clinical Journal of Chinese Medicine

基金：甘肃省科技厅自然基金(1308RJZA197)

语种：中文

中文关键词：特发性肺纤维化;水蛭素;纤溶酶原抑制剂-1

外文关键词：Idiopathic pulmonary fibrosis;Hirudin;Plasminogen inhibitor-1

摘要：目的:本研究选用天然水蛭素作为研究药物,采用RT-qPCR、Western blot方法检测IPF大鼠肺组织中PAI-1mRNA及蛋白表达,观察水蛭素干预大鼠IPF的作用,探讨其可能的作用机制,为临床运用水蛭治疗IPF提供理论依据,从而拓宽IPF的治疗途径。方法:选取180只SPF级同月龄健康的Wistar大鼠,随机数字表法抽取18只大鼠作为空白(K)组,剩余162只大鼠气管内注射博莱霉素统一造模,造模成功后随机选取108只分为模型(M)组、激素(J)组、水蛭素高剂量(G)组、水蛭素中剂量(Z)组、水蛭素低剂量(D)组、水蛭素中剂量联合激素(ZJ)组,每组18只,造模成功后第2天开始给药,分别在造模后7 d、14 d、28 d处死大鼠并收集标本,采用Western blot法检测大鼠肺组织中PAI-1的蛋白表达,RT-qPCR法检测PAI-1的基因表达。结果:不同时间点,M组大鼠肺组织PAI-1基因和蛋白表达均最强(P <0.01),而且随着时间的推移,表达强度逐渐增强,说明肺纤维化处于发展趋势,28天大鼠肺间质纤维化程度最重。各用药组进行比较发现:在7 d、14 d、28 d,ZJ组大鼠肺组织PAI-1基因和蛋白表达均最弱(P <0.01),G组次之(P <0.01),J组表达仅次于G组(P <0.01),J组表达显著低于Z组和D组(P <0.01),而Z组和D组相当(P> 0.05)。结论:本实验证明,水蛭素具有治疗大鼠肺间质纤维化的作用,通过降低PAI-1基因和蛋白表达水平从而起到治疗大鼠肺间质纤维化是其作用机理之一。同样发现,水蛭素的治疗作用有明显的剂量依赖性,而水蛭素联合激素治疗大鼠肺间质纤维化的疗效更佳。
Objective: To study the expression of PAI-1 mRNA and protein in lung tissue of rats with IPF by RT-qPCR and Western blot, and observe the effect of hirudin on IPF in rats. The mechanism of action provides a theoretical basis for the clinical application of leech treatment of IPF, thereby broadening the treatment of IPF. Methods: 180 healthy Wistar rats of SPF class were selected. 18 rats were randomly selected as blank (K) group. The remaining 162 rats were intratracheally injected with bleomycin to form a model. 108 rats were randomly divided into model (M) group, hormone (J) group, hirudin high dose (G) group, medium dose (Z) group, low dose (D) group, hirudin medium dose combined hormone (ZJ) groups, 18 rats in each group, were administered on the second day after successful modeling. The rats were sacrificed at 7 days, 14 days, and 28 days after model establishment, and the samples were collected. The protein of PAI-1 in rat lung tissue was detected by Western blot. Expression, gene expression of PAI-1 was detected by RT-qPCR. Results: At different time points, the expression of PAI-1 gene and protein in lung tissue of M group was the strongest (P<0.01), and the expression intensity increased gradually with time, indicating that pulmonary fibrosis is in the development trend;The degree of pulmonary interstitial fibrosis of 28-day rats is the heaviest. Comparing with the drug groups, the expression of PAI-1 gene and protein in the lung tissue of ZJ group was the weakest at 7 days, 14 days and 28 days (P<0.01), followed by G group (P<0.01). The expression of J group was second only to G group (P<0.01), and the expression of J group was significantly lower than that in the Z group and the D group (P<0.01), while Z group and D group were equivalent (P>0.05). Conclusion: hirudin has the effect of treating pulmonary interstitial fibrosis in rats. It is one of the mechanisms of action to treat pulmonary interstitial fibrosis in rats by reducing the expression level of PAI-1 gene and protein. It was also found that the therapeutic effect of hirudin was significantly dose-dependent, and that hirudin combined with hormones was more effective in treating pulmonary interstitial fibrosis in rats.