丹参联合川芎嗪对肺纤维化大鼠肺组织TGF-β_1表达及羟脯氨酸含量的影响

Effects of Combination of Salvia and Ligustrazine on the Expression of TGF-β_1 and Hydroxyproline in Lung Tissue of Pulmonary Fibrosis Rats

目的观察丹参联合川芎嗪注射液腹腔注射对博来霉素(BLM)致肺纤维化大鼠肺组织转化生长因子-β1(TGF-β1)表达及羟脯氨酸含量的影响。方法将90只成年雄性SD大鼠,随机分为6组:生理盐水(NS)组、BLM组、地塞米松(DM)组,中药干预组(A组、B组、C组)。NS组气管内灌注NS,BLM组、DM组及中药干预组(A组、B组、C组)气管内灌注BLM,随后NS组和BLM组每日腹腔注射NS,DM组每日腹腔注射DM,中药干预组(A组、B组、C组)每日给予不同剂量丹参和川芎嗪复合制剂腹腔注射,分别于气管内灌注药物后第7天,第14天和第28天处死每组大鼠各5只,左肺多聚甲醛固定后行HE染色、Masson染色进行肺泡炎及肺纤维化程度的判定;免疫组化法测定肺组织中TGF-β1的表达;右肺行肺组织羟脯氨酸含量的测定;血清用于检测肝肾功能。结果①NS组大鼠各时间点肺组织未发现肺泡间隔水肿、炎性细胞浸润及纤维化的形成。BLM组大鼠第7天时肺泡腔内可见大量炎性细胞浸润,第14天时炎症细胞减少,肺泡间隔内成纤维细胞显著增多,肺泡结构破坏,第28天时肺纤维化程度进一步加重,形成广泛纤维化。与BLM组比较,DM组大鼠各时间点肺泡炎及纤维化程度减轻(P<0.05)。A组大鼠肺泡炎和肺纤维化程度与BLM组类似(P>0.05)。B组、C组第14天肺泡炎及肺纤维化程度明显低于BLM组(P<0.05)。B组、C组第28天肺纤维化程度明显低于BLM组(P<0.05)。②与NS组比较,BLM组、DM组及中药干预组(A组、B组、C组)肺组织中TGF-β1的表达水平和羟脯氨酸含量明显升高(P<0.05)。③与BLM组比较,DM组及中药干预组(B组、C组)肺组织中TGF-β1的表达水平及羟脯氨酸含量明显降低(P<0.05)。④大鼠血清肝肾功能检测结果显示各组两两比较差异无统计学意义(P>0.05)。结论丹参联合川芎嗪腹腔注射能减轻BLM所致肺纤维化大鼠肺泡炎和肺纤维化程度,其机制可能与抑制肺组织TGF-β1的表达,减少羟脯氨酸含量有关。

Objective To investigate the effects of Salvia in combination with Ligustrazine on the expression of TGF-β1 and hydroxyproline in lung tissue of pulmonary fibrosis rats induced by bleomycin（BLM）.Methods 90 adult male SD rats were randomly divided into six groups：normal saline（NS） group,BLM group,dexamethasone（DM） group,Chinese crude drug groups（A group,B group,C group）.NS group was intratracheally instilled with NS,and BLM group,DM group and Chinese crude drug groups（A group,B group,C group） were intratracheally instilled with BLM.And then NS group and BLM group were intraperitoneally injected NS,DM group was intraperitoneally injected dexamethasone,and Chinese crude drug group（A group,B group,C group） were intraperitoneally injected different does of Salvia and Ligustrazine mixture.Five rats in each group were killed respectively on days 7,14,28 after intratracheal instillation.Left lung was fixed by paraform,and HE and Masson staining were performed to investigate alveolitis and fibrosis;TGF-β1 in lung tissue was assessed by immunohistochemistry;Hydroxyproline in the tissue of right lung was assessed;Serum was used to detect the liver and kidney function.Results ①Alveolar septal edema,inflammatory cell infiltration and fibrosis were not found in NS group.In the BLM group,lots of inflammatory cells infiltration were observed on days 7.Inflammatory cells were reduced,the number of the fibroblasts and matrix in alveolar septum were obviously increased,and alveolar structure was damaged on day 14.Pulmonary fibrosis was increased,severe fibrosis was found on day 28.Compared with BLM group,the degree of alveolitis and pulmonary fibrosis were significantly reduced in DM group at each time point（P〈0.05）,the degree of alveolitis and pulmonary fibrosis in the A group were similar to the BLM group（P〉0.05）.Compared with BLM group,the degree of alveolitis and pulmonary fibrosis in the B group and C group were remarkably ameliorated on day 14（P〈0.05）.Compared with BLM group,the degree of pulmonary fibrosis in the B group and C group remarkably reduced on day 28（P〈0.05）.②Both TGF-β1 expression and hydroxyproline content in lung tissue in BLM group,DM group and Chinese crude drug group（A group,B group,C group） were obviously increased than in the NS group（P〈0.05）.③Both TGF-β1 expression and hydroxyproline content in lung tissue in DM group and Chinese crude drug groups（B group,C group） were obviously lowered as compared to BLM group（P〈0.05）.④The liver and kidney function in each group had no significant difference（P〉0.05）.Conclusion Salvia and Ligustrazine mixture can alleviate alveolitis and fibrosis in BLM-induced pulmonary fibrosis rats,which might result from the inhibition of the expression of TGF-β1 and reduction of hydroxyproline content in lung tissue.