舒尼替尼、保肺定喘汤对慢性阻塞性肺疾病合并糖尿病小鼠肺血管重构的影响  

Effect of Sunitinib and Baofei Dingchuan Decoction on Pulmonary Vascular Remodeling in Chronic Obstructive Pulmonary Disease Complicating Diabetes Mice Model

作　　者：杨宏宽 王芳 张佳颖 潘俊杰 童卫泉 夏婷婷 陈芳[2] YANG Hong-kuan;WANG Fang;ZHANG Jia-ying;PAN Jun-jie;TONG Wei-quan;XIA Ting-ting;CHEN Fang(First Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou（310053;Respiratory PhysioL-ogy Laboratory,First Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou（310006)

机构地区：[1]浙江中医药大学第一临床医学院,杭州310053 [2]浙江中医药大学附属第一医院呼吸生理研究室,杭州310006

出　　处：《中国中西医结合杂志》2018年第9期1080-1086,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年资助项目(No.81302934)

摘　　要：目的探讨舒尼替尼、保肺定喘汤对慢性阻塞性肺疾病(COPD)合并糖尿病小鼠肺血管重构的影响及其是否通过调节低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)在肺组织的表达而发挥作用。方法将40只ob/ob小鼠随机分为4组:对照组、模型组、保肺定喘汤组和舒尼替尼组,每组10只。除对照组外,各组小鼠在自制熏烟箱内被动吸烟14周,并在第1、14天于气道内滴入脂多糖。造模结束后舒尼替尼组予苹果酸舒尼替尼胶囊30mg/(kg·d)灌胃,保肺定喘汤组予保肺定喘汤9.3g/(kg·d)灌胃,均干预2周。于第0、14、16周检测空腹血糖水平。肺组织HE染色,观察气道重构并测量小气道基底膜单位长度的管壁面积(WAt/Pbm),肺血管重构并测量管壁厚度与血管外径比(WT%)、管壁面积与血管总面积比(WA%);Masson染色观察支气管周胶原沉积并测量小气道基底膜单位长度的胶原面积(WAc/Pbm);CD31/α-SMA/DAPI免疫荧光染色观察肺血管平滑肌层变化。酶联免疫吸附测定法检测肺组织匀浆HIF-1α及VEGF含量。实时荧光定量PCR法检测肺组织HIF-1α、VEGF mRNA相对表达水平。结果与对照组比较,模型组第16周空腹血糖水平差异无统计学意义(P>0.05),WAt/Pbm、WAc/Pbm、WT%及WA%均增大(P<0.01),肺血管平滑肌层明显增厚,肺组织HIF-1α、VEGF蛋白表达和mRNA表达均增加(P<0.01)。与模型组比较,舒尼替尼组第16周空腹血糖水平降低(P<0.01),保肺定喘汤组差异无统计学意义(P>0.05);保肺定喘汤组及舒尼替尼组WAt/Pbm、WAc/Pbm、WT%及WA%均减小(P<0.01,P<0.05),肺血管平滑肌层重构均有所减轻,肺组织HIF-1α、VEGF蛋白表达和mRNA表达均减少(P<0.01,P<0.05)。结论舒尼替尼、保肺定喘汤可通过下调肺组织HIF-1α、VEGF表达进而减轻COPD合并糖尿病小鼠肺血管重构。Objective To observe the effect of Sunitinib and Baofei Dingchuan Decoction （BDD） on pulmonary vascular remodeling in chronic obstructive pulmonary disease（COPD） complicating diabe- tes mice model, and to study if its mechanism is relevant with the expression of hypoxia-inducible factor-1α（HIF-1α） and vascular endothelial grow factor（VEGF） in lung tissue. Methods Totally 40 ob/ob mice were randomly divided into 4 groups i.e,, the control group, the model group, the BDD group, and the sunitinib group, 10 in each group. All the mice except the control group were exposed to cigarette smoke in a homemade box for 14 weeks. At day 1 and 14, lipopolysaccharide（LPS） was dripped into the airway of the mice. After the model was established, mice in the sunitinib group were administered Sunitinib mal- ate at dosage of 30 mg/（ kg ~ d） by gastrogavage and mice in the BDD group were administered with BDD at dosage of 9.3 g/（kg ~ d） by gastrogavage, for consecutive 2 weeks. The levels of fasting blood glu- cose （FBG） was measured at week 0, 14 and 16. The airway remodeling was observed by HE staining and quantified by total bronchial wall area/basement membrane perimeter （WAt/Pbm）, the pulmonary vascular remodeling was quantified by vessel wall thickness /total vascular diameter（WT%） and vessel wall area /total vascular area（WA%）. The collagen deposition in peribronchus was observed by Masson staining and quantified by collagen wall area/basement membrane perimeter（WAc/Pbm）, the pulmonary vascular smooth muscle layer remodeling was observed by CD31/α-SMA/DAPI immunofluorescence stai- ning. The protein level of HIF-1 o~ and VEGF in lung tissue homogenate were detected by enzyme-linked im- munosorbent assay. Relative expressions of HIF-1α and VEGF mRNA in lung tissue were determined by real-time fluorescence quantitative PCR. Results Compared with the control group, the model group showed no significant difference in level of FBG at week 16 （P 〉0.05）, WAt/Pbm, WAc/Pbm, WT%

关 键 词：舒尼替尼 保肺定喘汤 慢性阻塞性肺疾病 糖尿病 肺血管重构 低氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R965[医药卫生—药理学]