从微观血瘀角度探讨补阳还五汤对特发性肺纤维化大鼠的影响

Study on the Effect of Buyang Huanwu Decoction on Idiopathic Pulmonary Fibrosis in Rats from the Perspective of Microscopic Blood Stasis

目的从微观血瘀角度探讨补阳还五汤对特发性肺纤维化(Idiopathic pulmonary fibrosis,IPF)大鼠的影响。方法将60只SD大鼠随机分为4组:对照组、模型组、尼达尼布组(0.1 g·kg^(-1))、补阳还五汤组(14.84 g·kg^(-1)),每组15只;采用气管内滴注博来霉素联合施加限制食量、睡眠剥夺刺激因素,建立气虚血瘀IPF大鼠模型;灌胃给药,每日1次,连续28 d。实验过程中观察大鼠一般生物学表征,对气虚血瘀症状进行量化评分;采用HE、Masson染色法观察肺组织病理变化,以Szapiel评分体系评价肺泡炎性程度,以Ashcroft评分体系评价大鼠肺组织纤维化情况;采用旷场实验检测大鼠行为学变化;采用血液流变学检测全血黏度指标;采用Image-Pro Plus 6.0软件分析计算HE染色切片中肺小动脉管壁厚度、肺小动脉管壁厚度与血管外径比值、管壁面积与血管总面积比值,以评价肺小动脉病理变化程度。结果与对照组比较,模型组大鼠气虚血瘀证候评分显著升高(P<0.01);肺组织结构紊乱,肺泡结构有不同程度破坏,肺泡腔及肺间质可见大量炎性细胞浸润,肺间质中可见明显的成纤维细胞聚集和大量的胶原沉积,Szapiel评分及Ashcroft评分均显著升高(P<0.01);第28天的运动总距离、进入中央区次数及中心格次数均显著减少(P<0.01);全血低切、中切和高切黏度及红细胞压积、聚集指数、电泳指数均显著升高(P<0.01);肺小动脉管壁厚度、肺小动脉管壁厚度与血管外径比值、管壁面积与血管总面积比值均显著增加(P<0.01),肺小动脉形态发生明显改变。与模型组比较,尼达尼布和补阳还五汤组大鼠的气虚血瘀证候评分显著下降(P<0.01);肺泡结构相对完整,炎性细胞浸润、胶原沉积和成纤维细胞聚积明显减轻,Szapiel评分及Ashcroft评分显著降低(P<0.01);第28天的运动总距离、进入中央区次数及中心格次数均显著增加(P<0.05,P<0.01);全血低切、中切和高切黏度及红细胞压积、聚集指数、电泳指数均明显下降(P<0.05,P<0.01);肺小动脉管壁厚度、肺小动脉管壁厚度与血管外径比值、管壁面积与血管总面积比值均显著降低(P<0.01)。结论补阳还五汤可显著改善IPF大鼠肺组织病理变化以及气虚血瘀状态,可能与降低全血黏度、抑制肺小动脉血管病变,从而增强肺功能有关。

Objective To investigate the effects of Buyang Huanwu Decoction(BYHWD)on idiopathic pulmonary fibrosis(IPF)rats from the perspective of microscopic blood stasis.Methods Sixty SD rats were randomly divided into four groups:control group,model group,Nintedanib group(0.1 g·kg^(-1)),and BYHWD group(14.84 g·kg^(-1)),with 15 rats in each group;a rat model of IPF with qi deficiency and blood stasis syndrome was established by intratracheal drip of Bleomycin(BLM)combined with food restriction and sleep deprivation stimuli;the drug was administered by gavage once daily for 28 consecutive days.The general biological characteristics of the rats were observed and the manifestations of qi deficiency and blood stasis syndrome were given quantitative assessment;lung histopathological changes were observed by HE and Masson staining,alveolar inflammation was evaluated by Szapiel scoring system and fibrosis was evaluated by Ashcroft scoring system;behavioral changes of the rats were detected by open field test;whole blood viscosity index was measured by blood rheology;Image-Pro Plus 6.0 software was used to analyze and calculate the wall thickness of small pulmonary arteries under HE staining,the ratio of wall thickness of small pulmonary arteries to outer diameter of blood vessels,and the ratio of wall area to total area of blood vessels,in order to evaluate the degree of pathological changes in small pulmonary arteries.Results Compared with the control group,the qi deficiency and blood stasis syndrome scores were significantly increased in the model group(P<0.01);the lung tissue structure was disturbed,with varying degrees of destruction of alveolar structure,a large number of inflammatory cells infiltrating the alveolar cavity and interstitium,and significant fibroblast aggregation and collagen deposition were seen in the interstitium;the Szapiel score and Ashcroft score were significantly increased(P<0.01);the total distance run,the number of entries into the central zone and the number of central compartments were significantly reduced on day 28(P<0.01);whole blood low-,medium-and high cut viscosities and erythrocyte pressure volume,aggregation index and electrophoresis index were significantly increased(P<0.01);the wall thickness of small pulmonary arteries,the ratio of wall thickness of small pulmonary arteries to the outer diameter of the vessel and the ratio of wall area to total vessel area were significantly increased(P<0.01).The morphology of small pulmonary arteries was significantly altered.Compared with the model group,the rats in the Nintedanib and BYHWD groups showed a significant decrease in qi deficiency and blood stasis syndrome scores(P<0.01);alveolar structure was relatively intact,inflammatory cell infiltration,collagen and fibroblast deposition were significantly reduced,and Szapiel and Ashcroft scores were significantly decreased(P<0.01);the total distance run,the number of times entering the central zone and the number of central compartments on day 28 were all significantly increased(P<0.05,P<0.01);whole blood low-,medium-and high-shear viscosity,hematocrit,aggregation index and electrophoretic index were significantly decreased(P<0.05,P<0.01);the wall thickness of pulmonary arterioles,the ratio of wall thickness to external diameter of pulmonary arterioles,and the ratio of wall area to total vascular area were significantly decreased(P<0.01).Conclusion BYHWD can significantly improved the histopathological changes of the lung and the state of qi deficiency and blood stasis in IPF rats,which may be related to the reduction of whole blood viscosity and inhibition of vascular lesions in small pulmonary arteries,thereby enhancing lung function.