血栓通注射液抗博来霉素致大鼠肺纤维化药效学研究

Pharmacodynamic study of Xueshuantong Injection on bleomycin induced pulmonary fibrosis in rats

目的研究血栓通注射液抗博来霉素诱导肺纤维化模型大鼠的药效学。方法将180只SD大鼠随机分为假手术组、模型组、吡非尼酮(阳性药,50 mg·kg^(-1))组和血栓通注射液低、中、高剂量(注射液原液1、2、4 mL·kg^(-1))组,每组30只,采用气管内注射博来霉素方法制备肺纤维化大鼠模型,于造模24 h后各给药组ip给药,并分别在造模7、14、28 d取材。观察大鼠一般状态、体质量、肺系数;应用DSI/BUXCO监测系统检测肺功能指标潮气量、气道阻力及肺顺应性变化;进行HE与Masson染色,光镜下观察肺组织病理变化;并采用酶联免疫吸附法(ELISA)检测大鼠肺组织中I型胶原(COL-Ⅰ)和纤维黏连蛋白(FN)的水平。结果通过对大鼠的一般情况观察,发现给予血栓通注射液后,肺纤维化模型大鼠的活动和精神状态均有改善。与模型组比较,吡非尼酮组、血栓通注射液组大鼠体质量显著增加(P<0.01、0.001),大鼠肺系数显著降低(P<0.05、0.01、0.001)。肺功能结果显示,与模型组比较,造模7 d时血栓通注射液中剂量显著上调动态肺顺应性(P<0.01);造模14 d时血栓通注射液高剂量显著降低气道阻力(P<0.05)和上调动态肺顺应性(P<0.01);造模28 d时吡非尼酮和血栓通注射液中剂量均显著上调动态肺顺应性(P<0.01)。HE及Masson结果显示,吡非尼酮组、血栓通注射液组的肺纤维化程度明显轻于模型组。ELISA结果显示,与模型组比较,造模7 d时吡非尼酮组、血栓通注射液高剂量组大鼠肺组织中的FN显著降低(P<0.01、0.001),吡非尼酮组COL-Ⅰ水平显著降低(P<0.001);造模14 d时吡非尼酮组和血栓通注射液中、高剂量组大鼠肺组织中的COL-Ⅰ水平显著降低(P<0.05、0.01),吡非尼酮组FN显著降低(P<0.05);造模28 d时吡非尼酮组和血栓通注射液中、高剂量组大鼠肺组织中的COL-Ⅰ、FN水平显著降低(P<0.05、0.01、0.001)。结论血栓通注射液可改善肺纤维化模型大鼠肺功能与肺部形态学病变,降低肺组织中COL-Ⅰ和FN水平,对肺纤维化大鼠产生保护作用。

Objective To investigate the effect of Xueshuantong Injection on bleomycin-induced pulmonary fibrosis model rats and its related mechanism.Methods Totally 180 SD rats were randomly divided into sham operation group,model group,pirfenidone group(10 mL·kg^(-1)),and Xueshuantong Injection low,medium,and high dose(injection stock solution 1,2,and 4 mL·kg^(-1))groups,with 30 rats in each group.The pulmonary fibrosis rat model was prepared by intratracheal injection of bleomycin.Xueshuantong Injection was injected ip in each group at 24 h after modeling,,and the materials were taken on the 7th,14th,and 28th days of modeling.The general state,body weight and lung coefficient of rats were observed.The changes of tidal volume,airway resistance and lung compliance were detected by DSI/BUXCO monitoring system.HE and Masson staining were performed to observe the pathological changes of lung tissues under light microscope.The levels of type I collagen(COL-Ⅰ)and fibronectin(FN)in serum were detected by enzyme-linked immunosorbent assay(ELISA).Results By observing the general situation of rats,it was found that after giving Xueshuantong Injection,the activity and mental state of pulmonary fibrosis model rats were improved.Compared with the model group,the body mass of rats in the pirfenidone group and Xueshuantong Injection group was significantly increased(P<0.01,0.001),and the lung coefficient of rats was lower than that in the model group(P<0.05,0.01,0.001).The results of pulmonary function showed that,compared with the model group,Xueshuantong Injection of medium dose significantly upregulated the dynamic lung compliance at seven days after modeling(P<0.01).At 14 days after modeling,Xueshuantong Injection of high dose significantly decreased airway resistance(P<0.05)and upregulated dynamic lung compliance(P<0.01).At 28 days after modeling,Xueshuantong Injection of medium dose and pirfenidone significantly increased lung compliance(P<0.01).The results of HE and Masson showed that the degree of pulmonary fibrosis in the pirfenidone group and Xueshuantong Injection group was significantly lighter than that in the model group.ELISA results showed that compared with the model group,FN in lung tissue of rats in pirfenidone group and Xueshuantong Injection high-dose group was significantly decreased at seven days after modeling(P<0.01,0.001),and the level of COL-Ⅰin pirfenidone group was significantly decreased(P<0.001).At 14 days after modeling,the COL-Ⅰlevel in lung tissue of rats in pirfenidone group and Xueshuantong Injection medium and high dose groups was significantly decreased(P<0.05,0.01),and FN in pirfenidone group was significantly decreased(P<0.05).The levels of COL-Ⅰand FN in lung tissue of rats in pirfenidone group and Xueshuantong Injection medium and high dose groups were significantly decreased at 28 days after modeling(P<0.01,0.001).Conclusion Xueshuantong Injection can improve pulmonary function and pulmonary morphological lesions in pulmonary fibrosis model rats,reduce the levels of Col-Ⅰand FN in lung tissue,and have a protective effect on pulmonary fibrosis rats.