芪丹颗粒剂对肺纤维化鼠I型前胶原和III型前胶原mRNA表达的影响

观察芪丹颗粒对博莱霉素致肺纤维化鼠的I型前胶原和III型前胶原mRNA表达的影响,探讨其作用机制。用160只SD大鼠按随机区组设计分为对照组(N)、模型组(M)、芪丹颗粒剂1组和2组(Q1、Q2)、氢化可的松1组和2组(H1、H2)共6组。对照组20只气管内灌注和灌胃均用生理盐水。余140只SD大鼠经气管内一次性灌注博莱霉素A5按BLM5mg·kg-1体重诱导大鼠肺间质纤维化,随机取40只为模型组。芪丹1组和氢可1组分别从造模后第2天灌注芪丹颗粒剂(3125mg/kg)和腹腔注射氢化可的松(25mg/kg),药物干预后第7、14、28天分别处死动物。芪丹2组和氢可2组分别从造模14天后灌注芪丹颗粒剂和腹腔注射氢化可的松(用量同前),2组动物第28、42天分别处死。用苏木素-伊红(HE)评价肺组织病理学变化和原位杂交方法检测各组大鼠肺I型和III型前胶原mRNA表达。肺组织病理HE染色显示芪丹组肺泡炎及肺纤维化程度均明显轻于模型组和氢可组(P<0.05)。在肺间质纤维化形成中,I型和III型前胶原mRNA表达呈动态变化,早期肺泡炎以III型前胶原mRNA大量增生为主,晚期纤维化期以I型前胶原mRNA增生为主。芪丹组III型前胶原mRNA的表达在第14天处于最高,至28天仍维持较高的水平,芪丹组III型前胶原mRNA的表达高于模型组和氢可组(P<0.05)。I型前胶原mRNA的表达在28天时芪丹1组和氢可1组及芪丹2组和氢可2组组间均有显著性差异,但在42天时,芪丹2组与氢化可的松2组其表达无显著差异(P<0.05)。大鼠肺纤维化的早期以III型前胶原mRNA的表达为主,晚期纤维化期以I型前胶原mRNA的大量表达为主。芪丹颗粒可减轻博莱要素诱导大鼠肺泡炎及肺纤维化的程度,其机制可能通过影响了I型和III型前胶原mRNA的代谢和表达,从而减慢肺间质纤维化的进程。芪丹颗粒对肺间质纤维化有治疗作用,且优于氢化可的松。

Effects of Qidan Granules on the Expression of Procollagen I and Procollagen III mRNA in Pulmonary Fibrosis Rats Induced by Bleomycin

To evaluate the expression of type I procollagen and type III procollagen mRNA by Qidan Granuless on Bleomycin induced pulmonary fibrosis in rats and to explore its pathogenesis, 160 SD rats in good conditions were randomly divided into six groups:normal control group, model group, Qidan Granules group(G1,G2) and hydrocortisonum group(H1,H2). Normal control group was treated with saline water. Pulmonary fibrosis was induced in rats by intratracheal instillation of Bleomycin (5 mg/kg body weight). 40 rats were chosen as model group and other rats were treated with Qidan Granules(3 125 mg/kg) or treated with Hydrocortisone(25 mg/kg)respectively. Rats of each group were sacrificed at 7, 14, 28 or 42 day post-Bleomycin and the lungs were obtained and fixed in paraformaldehyde and for in situ hybridization. All of the results were analysised by Leica Qwin V3 digital-image processing system. Qidan Granuless could obviously reduce alveolitis and fibrosis. III procollagen mRNA expression was increased in the acute alveolitis phrase of pulmonary fibrosis induced by Bleomycin, the expression of type I procollagen mRNA was mainly increased in the chronic stage of pulmonary fibrosis. The expression of III procollagen mRNA in Qidan Granules group was highest at 14 day and was higher than Hydrocortisone group(P<0.05). The expression of I procollagen mRNA was highest at 42 day, and there is a significant difference between Q1 and H1 group Q2 and H2 group at 28 day(P<0.05), but there is no significant difference between Q2 and H2 at 42 day. It is conclused Qidan Granules could obviously reduce alveolitis and fibrosis. Qidan Granules may alleviate the process of rat pulmonary fibrosis through influence the expression of I procollagen and III procollagen mRNA. The results strongly indicate the beneficial effects of Qidan Granules in protecting lung against Bleomycin-induced pulmonary fibrosis.