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盐酸班布特罗改善慢性阻塞性肺疾病模型大鼠的作用机制研究 Δ

杜孟姣 ，钟艳梅，邓善滨，余楚钦 ，陈莉诗，蔡延渠（广东药科大学新药研发中心/广东省药物新剂型重点实验
＊
#
室，广州 510006）

中图分类号 R285.5；R563.9 文献标志码 A 文章编号 1001-0408（2020）05-0564-07
DOI 10.6039/j.issn.1001-0408.2020.05.12

摘 要 目的：研究盐酸班布特罗对慢性阻塞性肺疾病（COPD）模型大鼠的改善作用机制，并寻找潜在的生物标志物。方法：将
30只大鼠随机分为正常组、模型组和盐酸班布特罗组（3.3 mg/kg），每组10只。模型组、盐酸班布特罗组大鼠采用烟熏联合脂多糖
滴注法建立COPD模型。造模成功后，盐酸班布特罗组大鼠灌胃相应药物，正常组和模型组大鼠灌胃等体积生理盐水，每天给药1
次，连续给药 45 d。末次给药后，采集各组大鼠血清样品及肺泡灌洗液，采用酶联免疫吸附法检测肺泡灌洗液中白细胞介素 6
（IL-6）、肿瘤坏死因子α（TNF-α）水平；采用液质联用（LC-MS）技术检测血清代谢物并进行代谢组学分析。采用正交偏最小二乘判
别分析（OPLS-DA）法筛选并鉴定差异代谢物，再结合相关文献找出潜在生物标志物，并利用MetPA分析平台对其进行代谢通路
富集分析。结果：与正常组比较，模型组大鼠肺泡灌洗液中IL-6、TNF-α水平显著升高（P＜0.05）；与模型组比较，盐酸班布特罗组
大鼠肺泡灌洗液中IL-6、TNF-α水平显著降低（P＜0.05）。代谢组学、OPLS-DA结果显示，共发现21个差异代谢物，其中12个为潜
在生物标志物（包括马来酰丙酮酸、羟基丙酮酸、羟基丙二酸半醛等），且盐酸班布特罗可显著降低马来酰丙酮酸、甲基硒代半胱氨
酸、5-脱氧-D-葡萄糖醛酸等标志物的水平（P＜0.05），升高羟基丙酮酸、羟基丙二酸半醛、丙二酸等标志物水平（P＜0.05）；生物标
志物主要富集于戊糖磷酸途径、乙醛酸和三羧酸代谢通路，其次富集于肌醇磷酸代谢通路、精氨酸与酪氨酸代谢通路和甘氨酸、丝
氨酸、苏氨酸代谢通路。结论：盐酸班布特罗改善COPD的机制可能与降低细胞调节因子IL-6、TNF-α水平有关，还可能与氨基酸
代谢、能量代谢、脂质代谢等通路有关。
关键词 盐酸班布特罗；慢性阻塞性肺疾病；机制；生物标志物；代谢组学；液质联用；大鼠

Study on the Mechanism of Bambuterol Hydrochloride in the Improvement of Chronic Obstructive
Pulmonary Disease Model Rats
DU Mengjiao，ZHONG Yanmei，DENG Shanbin，YU Chuqin，CHEN Lishi，CAI Yanqu（New Drug Research and
Development Center of Guangdong Pharmaceutical University & Guangdong Provincial Key Laboratory of New
Dosage Form，Guangzhou 510006，China）

ABSTRACT OBJECTIVE：To study the mechanism of Bambuterol hydrochloride in the improvement of chronic obstructive
pulmonary disease（COPD）model rats，and to find the potential biomarker. METHODS：Totally 30 rats were randomly divided
into normal group，model group and bambuterol hydrochloride group（3.3 mg/kg），with 10 rats in each group；COPD model was
established by lipopolysaccharide （LPS） infusion combined with smoking in model group and bambuterol hydrochloride group.
After modeling，bambuterol hydrochloride group was given relevant medicine intragastrically，normal group and model group were
given constant volume of normal saline intragastrically，once a day，for consecutive 45 d. After last medication，the serum sample
and alveolar lavage fluid of rats were collected. The levels of interleukin-6（IL-6）and tumor necrosis factor-α（TNF-α）in alveolar
lavage fluid were detected by ELISA. The serum metabolites were detected by LC-MS and analyzed by metabolomics. Orthogonal
partial least squares discriminant analysis（OPLS-DA）was used to screen out the differential metabolites. The potential biomarkers
were identified based on the related literature，and the metabolic pathway enrichment analysis was carried out by MetPA analysis
platform. RESULTS：Compared with normal group，the levels of IL-6 and TNF-α in alveolar lavage fluid of rats were increased
significantly in model group（P＜0.05）. Compared with model group，the levels of IL-6 and TNF-α in alveolar lavage fluid of rats
were decreased significantly in bambuterol hydrochloride group（P＜0.05）. Results of metabolomics and OPLS-DA showed that 21
differential metabolites and 12 potential biomarkers were found （including maleylpyruvate， hydroxypyruvate， tartronate
semialdehyde，etc.）. Bambuterol hydrochloride can significantly reduce the levels of maleylpyruvate，methylselenocysteine and
5-deoxy-D-glucuronic acid （P＜0.05）， while increase the
Δ 基金项目：国家自然科学基金资助项目（No.81503260）；广东省
levels of hydroxypyruvate， tartronate semialdehyde and
科技厅-广东省中医药科学院联合科研项目（No.2016A020226038）
malonic acid （P＜0.05）. These biomarkers were mainly
＊硕士研究生。研究方向：新药研发与转化。E-mail：15225955073
concentrated in pentose phosphoric acid pathway，glyoxylic
@163.com
# 通信作者：高级工程师，硕士生导师。研究方向：药物新剂型的 acid and tricarboxylic acid metabolism pathway，followed by
开发。E-mail：pn333@163.com inositol phosphoric acid metabolism pathway， arginine and

·564 · China Pharmacy 2020 Vol. 31 No. 5 中国药房 2020年第31卷第5期