摘要
目的观察红景天对肺纤维化大鼠肺组织I型胶原、转化生长因子β1(TGF-β1)及其信号通路分子Smad4表达的影响,探讨红景天对大鼠肺纤维化的防治作用与机制。方法健康SD大鼠40只随机分为4组:模型组(A组),正常对照组(B组),红景天14d治疗组(C组),红景天28d治疗组(D组)各10只。博莱霉素(BLM)气管内滴入建立大鼠肺纤维化模型,正常对照组气管内滴入生理盐水。在造模的第2日治疗组予红景天0.21g/kg灌胃干预,于第14,28日取肺组织,运用组织芯片结合图象分析技术,通过苏木素-伊红(HE)染色观察肺组织病理学改变,用免疫组化检测I型胶原蛋白和TGF-β1的表达,实时荧光定量RT-PCR检测I型胶原和TGF-β1的mRNA表达,原位杂交技术对Smad4 mRNA进行定位定量分析。结果红景天在两时相皆抑制I型胶原,TGF-β1的表达。C组I型胶原蛋白的IOD值为(0.362±0.241),D组I型胶原蛋白IOD值为(0.334±0.122),与A组(0.513±0.384)相比有统计学意义(P均<0.05)。C组TGF-β1蛋白的IOD值为(0.062±0.013),D组TGF-β1蛋白IOD值为(0.058±0.011),与A组(0.100±0.005)相比有统计学意义(P均<0.05)。C组I型胶原mRNA相对表达量为(0.041±0.017),D组I型胶原mRNA相对表达量为0.040±0.021,与A组0.071±0.054相比有统计学意义(P均<0.05)。C组TGF-β1 mRNA相对表达量为(0.0025±0.0013),D组TGF-β1 mRNA相对表达量为(0.0030±0.0005),与A组(0.0170±0.0157)相比有统计学意义(P均<0.05)。肺组织Smad4 mRNA表达呈局灶性,在炎症和胶原沉积部位表达较明显,而治疗组和模型组比较无统计学意义(P均>0.05);治疗组Smad4 mRNA表达量与正常对照组(0.165±0.099)比较亦无统计学意义(P>0.05)。结论红景天对大鼠肺纤维化具有一定治疗作用,其机制可能是通过抑制I型胶原及TGF-β1表达,从而干扰TGF-β-Smads信号通路对基因表达、蛋白合成的调节而实现的。
Objective To study the protective effect and mechanism of integripetal rhodiola herb（IRH） on rats with pulmonary fibrosis. Methods Forty SD rats were divided randomly into four groups （Group A： fibrosis model group, 10 rats. Group B：normal group, 10 rats. Group C：14 day -treatment group with IRH, 10 rats. Group D ：28 day -treatment group with IRH, 10 rats）. In the model group and the treatment group, pulmonary fibrosis was reproduced by the intratracheal injection of bleomycin （BLM）. In the normal control group, normal saline was given intratraeheally. IRH was administered intragastricly from day 1 till day 28 after modeling with a dose of 0.21 g/kg in the group C and D, while the normal saline was given intragastricly in the other groups. On the 29th day, rats were sacrificed. HE staining were undertaken after lung tissue was made into tissue microarray slices. Collagen Ⅰand transforming growth factor beta 1 （ TGF - β1 ） were tested by methods of immunohistochemical staining and real time quantitative PCR. Expression of Smad4 mRNA was tested by in situ hybridization. Results The expressions of Collagen Ⅰ in group C（0.362 ±0.241） and group D（0.334 ±0. 122） were much lower than that of group A （0.935 ±0.377,P〈0.05）. The expressions of TGF - β1 in group C （0.058 ± 0.011 ） and group D （ 0.062 ± 0.013 ） were much lower than that in group A （ 0.100 ±0.005,P〈0.05）. The expressions of Collagen Ⅰ mRNA in group C（0.041 ±0.017） and group D（0.040 ±0.021） were much lower than that of group A （ 0.071 ± 0.054 ,P 〈 0.05 ）. The expressions of TGF - β1 mRNA in group C （ 0. 0025 ± 0. 0013 ） and group D（0.0031 ±0.0005） were much lower than that in group A （0.0170 ±0.0157,P 〈0.05）. The IOD of Smad4 mRNA in treatment groups were no significant different from that in model group （P 〉 0.05 ）. Condusion Significant enhancement of the expressions of Collagen Ⅰ and TGF - β1 were observed in lung tissue of rats with pulmonary fibrosis. IRH has a preventive effect against pulmonary fibrosis, lnhibitting the expressions of pulmonary Collagen Ⅰ and TGF - β1 may be one of the mechanisms.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2007年第11期2772-2774,共3页
Lishizhen Medicine and Materia Medica Research
关键词
红景天
肺纤维化
Ⅰ型胶原
转化生长因子Β1
组织芯片
实时荧光定量PCR
Integripetal rhodiola herb
Pulmonary fibrosis
Collagen Ⅰ transforming growth factor beta 1
Tissue microar- ray
Real -time quantitative polymerase chain reaction（ real -time Q -PCR）
