迷迭香二萜芬提取物对肺纤维化大鼠肺组织TGF-β_1及其信号通路分子mRNA表达的影响  

Effects of Diterpene Phenol Extract of Rosmarinus Officinalis on TGFβ_1 and mRNA Expressions of Its Signaling Pathway Molecules in the Lung Tissue of Pulmonary Fibrosis Rats

作　　者：杨礼腾[1] 刘欣[2] 程德云[3] 方洵[3] 穆茂[3] 胡晓波[3] 聂莉[3] 

机构地区：[1]南方医科大学中西医结合医院呼吸内科,广州510315 [2]遵义医药高等专科学校中医系中医基础教研室,贵州遵义563000 [3]四川大学华西医院呼吸内科,成都610041

出　　处：《中国中西医结合杂志》2013年第6期819-824,共6页Chinese Journal of Integrated Traditional and Western Medicine

摘　　要：目的探讨迷迭香二萜芬提取物(diterpene phenol extract of Rosmarinus Officinalis,DERO)调控肺纤维化肺胶原代谢失衡的作用机制。方法 50只健康SD大鼠,随机分为生理盐水组(NS组)、博莱霉素肺损伤组(BLM组)及DERO低、中、高剂量[50、100、200mg/(kg·d)]组(分别简称为DERO1、2、3组),每组10只,通过博莱霉素一次性气管内滴入复制肺纤维化大鼠模型,并在肺损伤修复过程中予DERO灌胃干预,于第29天早上取大鼠肺组织,运用组织芯片,进行HE、胶原纤维染色及免疫组化、原位杂交,分别检测肺组织有核细胞数、胶原蛋白、Ⅰ型胶原(typeⅠcollagen,Collagen-Ⅰ)和转化生长因子βⅡ型受体(transforming growth factor-beta typeⅡreceptor,TGFβRⅡ)、Smad4mRNA的表达,应用实时荧光定量RT-PCR技术检测转化生长因子beta1(transforming growth factor-beta1,TGF-β1)mRNA的表达。结果与NS组比较,BLM组肺组织胶原沉积明显,炎症浸润程度较重(P<0.05,P<0.01),DERO1组与BLM组比较,两项指标差异无统计学意义(P>0.05),DERO2及DERO3组肺组织胶原沉积及炎症浸润程度皆明显减轻(P<0.05,P<0.01);与NS组比较,BLM组肺组织Collagen-Ⅰ、TGF-β1RⅡ、Smad4mRNA及TGF-β1 mRNA表达明显上调(P<0.05,P<0.01),与BLM组比较,DERO1组4项指标变化不明显(P>0.05),而DERO2及DERO3组,Collagen-Ⅰ、TGF-β1RⅡ及TGF-β1mRNA表达皆有明显下调(P<0.05,P<0.01),但两组Smad4mRNA下调不明显(P>0.05),与DERO1组比较,DERO2及DERO3组除Smad4mRNA外,余3项指标皆低于DERO1组(P<0.05),而DERO2组与DERO3组比较,4项指标差异无统计学意义(P>0.05)。结论 DERO在肺纤维化胶原代谢失衡中具有调控作用,能抑制胶原纤维的过度沉积,尤其是Collagen-Ⅰ的过度沉积,其作用机制可能主要是通过抑制TGF-β1、TGFβRⅡmRNA表达的上调,从而干扰TGF-β-Smad信号通路对靶基因尤其是Ⅰ型前胶原靶基因的激活而实现的。Objective To investigate the regulative mechanism of the diterpene phenol extract of Rosmarinus Officinalis （DERO）on the imbalance of collagen metabolism of the tung tissue in pulmonary fibrosis rats. Methods Fifty healthy Sprague-Dawley rats were randomly divided into the normal saline group （NS）, the bleomycin-induced lung injury group （BLM）, the low dose DERO group （at the daily dose of 50 mg/kg）, the moderate dose DERO group （at the daily dose of 100 mg/kg）, and the high dose DERO group （at the daily dose of 200 mg/kg）, 10 in each group （abbreviated as DERO 1,2,3, respec-tively）. The pulmonary fibrosis rat model was prepared by disposable intratracheal instillation of bleomy- cin. DERO was administered by gastrogavage as intervention during the repairing process of lung injury. On the morning of the 29th day, the rats＇ lung tissue was extracted. The karyocyte number, collagen pro- tein, type Ⅰ collagen （collagen Ⅰ ） and transforming growth factor-beta type Ⅱ receptor（TGFβR Ⅱ ）, Smad4 mRNA expressions were semi-quantitatively determined using tissue microarray, HE staining, collagen fiber dyeing, immunohistochemical assay, and in situ hybridization. Using real-time fluorescent quantification RT-PCR, the mRNA expression of transforming growth factor-beta1 （TGF-β1） were detec- ted. Results Compared with the NS group, the collagen deposition of the lung tissue was obvious and the inflammatory infiltration was more severe in the BLM group （P 〈0.05, P 〈0.01 ）. There was no statis- tical difference in the aforesaid 4 indices between the DERO1 group and the BLM group （P 〉0.05）. The collagen deposition and the inflammatory infiltration were obviously alleviated in the DERO2 and DERO3 groups （P 〈0.05, P 〈0.01 ）. Compared with the NS group, the mRNA expressions of collagen- Ⅰ , TGF-β1 RⅡ, Smad4, and TGF-β1 were obviously up-regulated in the BLM group （P 〈0.05, P 〈0.01 ）. Compared with the BLM group, the aforesaid four indices were no

关 键 词：肺纤维化 转化生长因子beta1 迷迭香 胶原 

分 类 号：R285.5[医药卫生—中药学]