电针对慢性阻塞性肺疾病小鼠D-多巴色素互变异构酶的影响  

Effect of Electroacupuncture on D-Dopachrome Tautomerase in Mice with Chronic Obstructive Pulmonary Disease

作　　者：刘海燕 吴慧慧 潘祝 项水英 张新芳[2] 王媛 刘自兵[1,2] LIU Haiyan;WU Huihui;PAN Zhu;XIANG Shuiying;ZHANG Xinfang;WANG Yuan;LIU Zibing(College of Acupuncture and Moxibustion,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;School of Integrated Traditional Chinese and Western Medicine,Anhui University of Chinese Medicine,Anhui Hefei 230012,China)

机构地区：[1]安徽中医药大学针灸推拿学院,安徽合肥230012 [2]安徽中医药大学中西医结合学院,安徽合肥230012

出　　处：《安徽中医药大学学报》2024年第6期48-54,共7页Journal of Anhui University of Chinese Medicine

基　　金：安徽省自然科学基金项目(2008085MH267);安徽高校自然科学研究重大项目(2022AH040078);安徽高校自然科学研究重点项目(KJ2021A0569,KJ2021A0599)。

摘　　要：目的探讨D-多巴色素互变异构酶(D-dopachrome tautomerase,DDT)在电针“肺俞”“足三里”穴治疗慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)小鼠中的作用机制。方法将50只C57小鼠随机分为空白组、模型组、电针组、mDDT-191组、mDDT-191+电针组,每组10只。采用香烟烟雾暴露联合脂多糖鼻腔滴入法复制COPD小鼠模型。采用肺功能仪检测小鼠肺功能,苏木精—伊红染色法观察小鼠肺组织形态,ELISA法检测小鼠血清及支气管肺泡灌洗液中DDT、肺泡表面活性蛋白-C(surface active protein-C,SP-C)、肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6水平,实时定量PCR及Western blot法检测肺组织DDT、SP-C mRNA及其蛋白表达水平,免疫组织化学法观察小鼠肺组织DDT、SP-C表达水平。结果与空白组比较,模型组小鼠用力肺活量(forced vital capacity,FVC)、第0.05秒用力呼气量(forced expiratory volume in 0.05 second,FEV_(0.05))、第0.1秒用力呼气量(forced expiratory volume in 0.1 second,FEV0.1)、FEV_(0.05)/FVC、FEV0.1/FVC、SP-C免疫反应阳性细胞平均吸光度值、肺组织SP-C mRNA及其蛋白表达水平均显著降低(P<0.05),肺组织炎症细胞浸润明显,各炎症因子及DDT的表达水平均显著升高(P<0.05)。与模型组比较,mDDT-191组肺功能各项指标明显改善(P<0.05),炎症细胞浸润减轻,各炎症因子及DDT表达水平显著下降(P<0.05),SP-C表达水平显著升高(P<0.05);与mDDT-191组比较,mDDT-191+电针组肺功能各项指标显著改善(P<0.05),各炎症因子及DDT表达水平显著下降(P<0.05),SP-C表达水平显著升高(P<0.05)。结论DDT促进COPD炎症反应;电针治疗COPD小鼠能降低DDT及升高SP-C表达水平,其在改善呼吸功能、减轻肺部炎症反应、修复肺泡细胞功能方面,与DDT小干扰RNA有协同作用。Objective To investigate the mechanism of action of D-dopachrome tautomerase(DDT)in the treatment of mice with chronic obstructive pulmonary disease(COPD)by electroacupuncture at Feishu and Zusanli points.Methods A total of 50 C57 mice were randomly divided into blank group,model group,electroacupuncture group,mDDT-191 group,and mDDT-191+electroacupuncture group,with 10 mice in each group.Cigarette smoke exposure combined with intranasal administration of lipopolysaccharide was used to establish a mouse model of COPD.The lung function of mice was assessed;HE staining was used to observe the morphology of lung tissue;ELISA was used to measure the levels of DDT,surface active protein-C(SP-C),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in serum and bronchoalveolar lavage fluid;quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of DDT and SP-C in lung tissue,and immunohistochemistry was used to measure the expression levels of DDT and SP-C in lung tissue.Results Compared with the blank group,the model group had significant reductions in forced vital capacity(FVC),forced expiratory volume at 0.05 seconds(FEV 0.05),forced expiratory volume in 0.1 second(FEV 0.1),FEV 0.05/FVC,FEV 0.1/FVC,the average optical density of cells with positive SP-C immune response,and the mRNA and protein expression levels of SP-C in lung tissue(P<0.05),with significant inflammatory cell infiltration in lung tissue and significant increases in the expression levels of various inflammatory factors and DDT(P<0.05).Compared with the model group,the mDDT-191 group had significant improvements in pulmonary function parameters(P<0.05),alleviation of inflammatory cell infiltration,significant reductions in the content of inflammatory factors and the expression level of DDT(P<0.05),and a significant increase in the expression level of SP-C(P<0.05).Compared with mDDT-191 group,the mDDT-191+electroacupuncture group had significant improvements in pulmonary function

关 键 词：慢性阻塞性肺疾病 D-多巴色素互变异构酶 电针 细胞表面活性蛋白-C 

分 类 号：R245.3[医药卫生—针灸推拿学]