健脾益肺Ⅱ号方对烟草烟雾暴露联合脂多糖气道滴注诱导大鼠膈肌损伤的影响  

Effect of Jianpi Yifei Formula Ⅱ on Diaphragm Injury Induced by Cigarette Smoke Exposure Combined with Lipopolysaccharide Intratracheal Instillation

作　　者：于旭华 胡涛[2] 陈远彬 吴蕾 范龙 伍思亮 许银姬 林琳 YU Xu-hua;HU Tao;CHEN Yuan-bin;WU Lei;FAN Long;WU Si-liang;XU Yin-ji;LIN Lin(Department of Pulmonary and Critical Care Medicine,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120;Department of Traditional Chinese Medicine,Guangzhou First People’s Hospital,Guangzhou 510120)

机构地区：[1]广州中医药大学第二附属医院呼吸与危重症科,广州510120 [2]广州市第一人民医院中医科,广州510120

出　　处：《中国中西医结合杂志》2020年第7期829-835,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81373566,No.81573895,No.81673897,No.81603554)。

摘　　要：目的探讨健脾益肺Ⅱ号方对烟草烟雾暴露联合脂多糖(LPS)气道滴注大鼠膈肌的保护作用及其机制。方法58只8周龄SD大鼠,根据出生时间均衡分配为正常组、模型组、健脾益肺Ⅱ号高剂量组[20 g/(kg·d)]、健脾益肺Ⅱ号低剂量组[10 g/(kg·d)]。模型组及药物干预组给予熏烟12周,并于熏烟第7、21天分别滴注200μL LPS。正常组暴露于空气,气道滴注生理盐水。用药组于第9周开始灌胃给药,正常组及模型组给予相应生理盐水灌胃,至第12周。记录并比较大鼠体重变化;HE染色方法观察肺部病理和膈肌病理改变;检测跨膈压和膈肌肌电;免疫组化法观察膈肌中Bax表达;通过qPCR方法检测泛素蛋白酶体通路C2蛋白亚基、E2/14k、MRuf-1、MAFbx mRNA表达。结果与正常组比较,模型组大鼠体重增长较少(P<0.01),单位视野内肺泡个数减少(P<0.01),跨膈压增高(P<0.05),膈肌炎症细胞浸润增多,Bax蛋白表达和MRuf-1 mRNA表达增多(P<0.01);与模型组比较,健脾益肺Ⅱ号高、低剂量组大鼠体重增长未见增加(P>0.05),健脾益肺Ⅱ号高剂量组大鼠肺泡数量明显增多,跨膈压明显降低(P<0.05);膈肌纤维间炎症细胞的浸润减少;膈肌组织中Bax的蛋白表达减少(P<0.01);健脾益肺Ⅱ号高、低剂量组膈肌组织中泛素E3连接酶MRuf-1的mRNA表达降低(P<0.01)。结论健脾益肺Ⅱ号方可减少膈肌细胞凋亡,其机制可能与减轻模型大鼠肺泡扩张程度和胸腔压力,减少膈肌炎症浸润和抑制泛素—蛋白酶体通路的激活有关。Objective To investigate the protective effect and mechanism of Jianpi Yifei FormulaⅡ(JYⅡ)on diaphragm injury induced by cigarette smoke exposure combined with lipopolysaccharide(LPS)intratracheal instillation.Methods Totally 58 SD rats aged 8 weeks were divided into control group,model group,high dose JYⅡgroup(20 g·kg-1·d-1),and low dose JYⅡgroup(10 g·kg-1·d-1)based on the birth date.Rats in model group and medicated groups were exposed to cigarette smoke for 12 weeks,and were intratracheal-instilled with 200μL LPS at day 7 and 21.Rats in control group were exposed to the air,and were intratracheal-instilled with saline instead.Rats in medicated groups were administered with JYⅡby gastrogavage from week 9 to week 12.Equal volume of normal saline was given by gastrogavage to rats in control group and model group.Body weight changes were recorded and compared before culling.Histology of lungs and diaphragm were observed using HE stain.Transdiaphragmatic pressure and electromyography were tested.Bax expression was tested using immumohistochemical staining.The mRNA expressions of C2,E2/14K,MRuf-1,MAFbx in Ub-proteasome pathway were detected by qPCR.ResultsCompared with the control group,body weight gain was significantly decreased(P<0.01),the number of pulmonary alveolus decreased(P<0.01),and transdiaphragmatic pressure increased(P<0.01)in the model group.Additionally,inflammatory cells influx,Bax protein expression and MRuf-1 mRNA expression in diaphragm increased in the model group(P<0.01).Compared with the model group,body weight gain was not changed in both JYⅡgroups(P>0.05),the number of pulmonary alveolus were increased(P<0.05),transdiaphragmatic pressure(P<0.05),inflammatory cells influx,and Bax protein expression decreased in high dose JYⅡgroup(P<0.01).The mRNA expression of MRuf-1 suppressed in diaphragm decreased in both JYⅡgroups(P<0.01).Conclusion JYⅡcould reduce apoptosis of skeletal muscle cells in diaphragm,and its mechanism might reduce pressure in chest,alleviate influx of inf

关 键 词：健脾益肺Ⅱ号 慢性阻塞性肺病 膈肌 

分 类 号：R285.5[医药卫生—中药学]