The Effect Of M1/M2 Macrophage Polarization Regulation By The Yiqi Huatan Quyu Formula On Lung Inflammation And Vascular Remodeling In Chronic Obstructive Pulmonary Disease Rats

ObjectiveTo investigate the effects of Yiqi Huatan Quyu formula to regulate macrophage polarization on lung inflammation and vascular remodeling in COPD rats,and to provide new ideas for the clinical treatment and research of COPD.MethodsTwo animal models of COPD progressing to pulmonary hypertension（hereafter referred to as"COPD rats"）were simulated using cigarette smoke（Cigarette Smoke Extracts,CSE）,hypoxia and monocrotaline（MCT）as exposure factors.In this case,CSE and hypoxia were combined to create the first animal model.The second animal model was completed by CSE and MCT.Eight groups consisted of 48 SPF-rated male rats,and the rats were divided into Control,Hypoxia,Hypoxia-HYQ（Hyp-HYQ）and Hypoxia-Ligustrazine（Hyp-LIG）groups as well as Control,MCT,MCT-HYQ and MCT-Ligustrazine（MCT-LIG）groups（n=6 in each group）.Lung function of the rats was measured using the Lung Function Analysis System.Co-expression of macrophage markers CD68,CD86 and CD206 was measured by immunofluorescence.WB was used to detect the expression levels of macrophage markers CD68,CD86,INOS,CD206 and CD163 proteins.ELISA was used to detect the cytokines IL-6,IL-1β,TNF-α,IL-10,b FGF and VEGF expression levels.Results（1）Changes in lung function:Compared with Hypoxia group and MCT group,the levels of FEV_0.3,FVC and FEV_0.3/FVC%in rats in Hyp-YHQ group and MCT-YHQ group increased,and the difference was statistically significant（p<0.05）.（2）Immunofluorescent protein expression in the lungs:pathologically,compared with the Hypoxia and MCT groups,the destruction of alveolar wall structure in the lung tissue of rats in the Hyp-YHQ and MCT-YHQ groups was alleviated,and inflammatory cell infiltration in all layers of the tubular wall was significantly reduced,and the thickness of the pulmonary vessels became thinner.In addition,the expression of immunofluorescence-labeled M1 marker CD86 was significantly higher in the Hypoxia and MCT groups than in the Hyp-YHQ and MCT-YHQ groups,while the M2marker CD206 showed the opposite result.（3）Macrophage-related protein expression in the lungs:compared with the Hypoxia and MCT groups,the expression of CD68,M1 marker CD86 and i NOS was reduced in the lungs of the Hyp-YHQ and MCT-YHQ groups,while the expression of M2markers CD206 and CD163 was significantly increased（p<0.05）.（4）Serum expression of serum factors related to pulmonary vascular inflammation and remodeling:compared with the Hypoxia and MCT groups,the levels of IL-6,IL-1βand TNF-α,which are regulated by M1-type macrophages,were down-regulated in the serum of rats in the Hyp-YHQ and MCT-YHQ groups,while IL-10,which is involved in the regulation of M2-type macrophages,was significantly up-regulated（p<0.05）.And VEGF and b FGF,which are associated with vascular remodelling,were significantly decreased（p<0.05）.ConclusionTwo animal models of slow-onset lung combined with progressive pulmonary arterial hypertension prepared by CSE combined with hypoxia or CSE combined with MCT showed that Yiqi Huatan Quyu formula can improve lung inflammation and vascular remodelling in COPD rats by regulating the polarisation of AMs from M1 to M2 types,inducing the expression of anti-inflammatory proteins and cytokines,and reducing the release of pro-inflammatory factors and related proteins.