摘要
目的:探讨穿心莲内酯灌胃对博来霉素(BLM)致肺纤维化大鼠支气管肺泡灌洗液(BALF)中TNF-α、TGF-β1浓度和肺组织Ⅰ、Ⅲ型胶原mRNA表达的影响。方法:取健康雄性SD大鼠90只,随机分为生理盐水(NS)组、BLM组、泼尼松(Pred)组、不同剂量穿心莲内酯组(即穿A组62.5mg/kg、穿B组125 mg/kg、穿C组250 mg/kg),各组大鼠15只,分别气管内灌注BLM(BLM组、Pred组、穿A组、穿B组、穿C组)或NS(NS组)后,每天给予NS(NS组、BLM组)、Pred(Pred组)或穿心莲内酯(穿A组、穿B组、穿C组)灌胃,各组分别于气管内灌注药物后第7、14、28天处死大鼠5只。用HE、Masson染色观察肺泡炎症和纤维化改变;实时荧光定量逆转录-聚合酶链反应检测肺组织Ⅰ、Ⅲ型胶原mRNA表达;酶联免疫吸附试验测定BALF中TNF-α、TGF-β1浓度;同时监测肾功能指标血尿素氮(BUN)、肌酐(Cr)及肝功能指标血丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)。结果:肝肾功能监测显示:不同剂量穿心莲内酯组、NS组、BLM组、Pred组所监测的AST、ALT、BUN、Cr比较,差异无统计学意义(P>0.05)。NS组大鼠肺组织未发现肺泡间隔水肿、炎性细胞浸润和纤维化形成。BLM组第7天时肺泡腔内可见大量炎性细胞浸润,第14天时肺泡炎仍存在,但炎症细胞明显减少,肺泡间隔内成纤维细胞明显增多,肺泡结构破坏,肺泡隔增宽,第28天时炎症较前减轻,肺纤维化程度加重,部分肺泡腔消失,形成严重纤维化。穿A组病理形态改变与BLM组相似。Pred组、穿B组、穿C组大鼠第7天有较多的炎症细胞浸润及局部聚积,第14天和第28天的纤维化病理改变均较BLM组、穿A组明显减轻。NS组各个时间点BALF中TGF-β1、TNF-α含量均明显低于同时间点BLM组、Pred组、穿A组、穿B组、穿C组BALF中的浓度(P<0.05)。BLM组3个时间点BALF中TGF-β1、TNF-α含量较Pred组、穿B、穿C组高(P<0.05)。与BLM组比较,穿A组BALF中TGF-β1、TNF-α含量无统计学意义。NS组各个时间点肺组织Ⅰ、Ⅲ型胶原mRNA表达均明显低于同时间点BLM组、Pred组、穿A组、穿B组、穿C组肺组织中的表达(P<0.05)。BLM组3个时间点Ⅰ、Ⅲ型胶原mRNA表达较Pred组、穿B组、穿C组高(P<0.05)。与BLM组比较,穿A组肺组织Ⅰ、Ⅲ型胶原mRNA表达无统计学差异。结论:穿心莲内酯灌胃可减轻BLM致肺纤维化大鼠肺泡炎和肺纤维化程度,降低肺组织Ⅰ、Ⅲ型胶原mRNA表达,降低BALF中TNF-α、TGF-β1浓度,且对肝肾无明显毒副作用。
AIM：To investigate the effects of andrographolide on the concentration of TNF-α and TGF-β1 in bronchoalveolar lavage fluid（BALF） and the expressions of type Ⅰand Ⅲ collagen mRNA in Lung tissue in bleomycin（BLM）-induced pulmonary fibrosis in rats.METHODS：90 healthy SD male rats were randomly divided into 6 groups with 15 rats each group：normal saline（NS） group,BLM group,prednisone（Pred） group and different doses of andrographolide groups（andrographolide group A 62.5 mg/kg,andrographolide group B 125 mg/kg and andrographolide group C 250 mg/kg）.BLM was given to BLM group,Pred group and andrographolide group A,B,C by intratracheal instillation,and same volume of NS was given to NS group in the same way.And then NS was given to NS group and BLM group,Pred was given to Pred group and different does of andrographolide were given to andrograoholide group A,B,C by gavage every day.Five rats of each group were killed respectively at day 7,14,28 after intratracheal instillation.Alveolitis and fibrosis were observed by HE and Masson staining.Real-time fluorescent quantitative reverse transcription-polymerase chain reaction（FQ RT-PCR） was performed to detect the expressions of type Ⅰand Ⅲ collagen mRNA in lung tissue,and the concentration of TNF-α and TGF-β1 in BALF was determined by enzyme-linked immunosorbnent assay.Blood urea nitrogen（BUN）,creatinine（Cr）,alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were also examined.RESULTS：（1） The AST,ALT,BUN and Cr in every group had no significant diference（P0.05）.（2） Alveolar septal edema,inflammatory cell infiltration and fibrosis were not found in NS group.In the BLM group,lots of inflammatory cells infiltration were observed in the alveolar at day 7;the alveolitis was still existed,but inflammatory cells were significantly reduced,and the number of the fibroblasts and matrix in alveolar septum were obviously increased at day 14,at the same time,alveolar structure was damaged and alveolar septum widened;the inflammation cells infiltration of the alveolar was relieved,pulmonary fibrosis was increased,and parts of alveolar space was disappeared,severe fibrosis was found at day 28.It was similar between andrographolide group A and BLM group in pathomorphology.A lot of inflammatory cells infiltration and local accumulation were observed at day 7 in andrographolide group B,C and Pred group.However,compared with andrographolide group A and BLM group,the fibrosis at day 14,28 was significantly reduced.（3） The concentration of TGF-β1,TNF-α in BALF of NS group was significantly lower than that of Pred group,BLM group,andrographolide group A,B,C at each time point（P0.05）.The concentration of TGF-β1 and TNF-α in BALF of BLM group at day 7,14,28 was higher than that of Pred group,andrographolide group B and andrographolide group C（P0.05）.Compared with BLM group,the concentration of TGF-β1 and TNF-α in BALF of andrographolide group A had no significant difference.（4） The expression of type Ⅰand Ⅲ collagen mRNA in lung tissue of NS group was significantly lower than that of group Pred,BLM,andrographolide group A,B,C at each time point（P0.05）.The expression of type Ⅰand Ⅲ collagen mRNA in lung tissue of BLM group at day 7,14,28 was higher than that of Pred group,andrographolide group B and andrographolide group C（P0.05）.Compared with BLM group,the expression of type Ⅰand Ⅲ collagen mRNA in lung tissue of andrographolide group A had no significant difference.CONCLUSION：In BLM-induced rat pulmonary fibrosis,andrographolide could attenuate alveolitis and fibrosis,decrease mRNA expression of collagen Ⅰ and Ⅲ in lung tissue and decrease the concentration of TNF-α and TGF-β1 in BALF.It had no side effects on liver and kidney function.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2011年第7期725-729,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
四川省杰出青年基金资助项目(2009-04-395)
泸州市科技局资助项目(07070)
