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浙江大学学报（医学版）
浙江大学学报（医学版）  2022, Vol. 51 Issue (1): 53-61    DOI: 10.3724/zdxbyxb-2021-0203
中药现代化     
清肺口服液在特发性肺纤维化中的治疗作用及网络药理学研究
张轶雯,盛孔胜,宋飞凤,潘宗富,邹小舟,刘宇佳,黄萍()
杭州医学院附属人民医院 浙江省人民医院临床药学中心药学部， 浙江 杭州 310014
Efficacy of Qingfei oral liquid for idiopathic pulmonary fibrosis in rats and related network pharmacology study
ZHANG Yiwen,SHENG Kongsheng,SONG Feifeng,PAN Zongfu,ZOU Xiaozhou,LIU Yujia,HUANG Ping()
Department of Pharmacy, Clinical Pharmacy Center, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, Hangzhou 310014, China
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摘要：

目的：考察清肺口服液在特发性肺纤维化中的治疗作用及机制。方法：72只SD大鼠分为正常对照组、模型对照组、吡非尼酮组（给予吡非尼酮 50?mg·kg–1·次–1，每天2次）、清肺口服液组（给予清肺口服液3.6?mL/kg，每天1次）处理21?d，获取肺组织进行HE染色、马森染色和转化生长因子-β（TGF-β）免疫组织化学染色，其余组织进行匀浆后检测超氧化物歧化酶（SOD）、丙二醛（MDA）和谷胱苷肽（GSH）。采用网络药理学方法，利用BATMAN-TCM数据库检索清肺口服液组成药物的化学成分及其对应靶点，随后构建成分-靶点-疾病网络图，进行通路富集分析。结果：组织病理学结果提示，清肺口服液给药后可改善肺纤维化状态，且效果与吡非尼酮类似；清肺口服液可减少肺组织TGF-β表达，对SOD、MDA和GSH也有调控作用。成分-靶点-疾病网络构建及基因功能注释结果显示，清肺口服液与调控炎症、细胞外基质和细胞因子等通路相关。结论：清肺口服液可能通过调控炎症等信号通路发挥抗特发性肺纤维化作用。
关键词： 特发性肺纤维化;  清肺口服液;  网络药理学;  氧化还原;  转化生长因子-β;  大鼠    
Abstract:

Objective:To investigate the therapeutic effect and mechanism of Qingfei oral liquid in idiopathic pulmonary fibrosis. Methods: Seventy-two male SD rats were divided into control group, model group, pirofenidone group and Qingfei group with 18 animals in each group. The idiopathic pulmonary fibrosis was induced in last three groups by intratracheal injection of 3?mg/kg bleomycin; pirofenidone group was given oral administration of 50?mg/kg pirofenidone b.i.d for 21?d, and Qingfei group was given Qingfei oral liquid 3.6?mL/kg q.d for 21?d. Lung tissues were obtained for HE staining, Masson staining and transforming growth factor (TGF)-β immunohistochemical staining. Superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) were detected in tissue homogenates. The BATMAN-TCM database was used to retrieve the chemical components and their corresponding targets of Qingfei oral solution by network pharmacology method, and then the component-target-disease network diagram was constructed. Finally, the pathway enrichment analysis was carried out to explore the molecular mechanism of Qingfei oral liquid against idiopathic fibrosis. Results: Histopathology results showed that Qingfei oral liquid had a similar relieving effect on pulmonary fibrosis as the positive drug pirfenidone; TGF-β secretion had a significant reduction in lung tissues of Qingfei group; and Qingfei oral liquid had better regulatory effect on SOD, MDA and GSH than pirfenidone. The results of component-target-disease network and pathway enrichment analysis showed that the related molecular pathways were concentrated in inflammation, extracellular matrix and cytokines. Conclusion: Qingfei oral liquid has a good therapeutic effect on idiopathic pulmonary fibrosis in rats via regulation of inflammation, extracellular matrix and cytokines.
Key words: Idiopathic pulmonary fibrosis    Qingfei oral liquid    Network pharmacology    Oxidation-reduction    Transforming growth factor-β    Rats
收稿日期: 2021-07-18 出版日期: 2022-05-17
CLC:  R965  
基金资助: 浙江省医药卫生科技计划（2018ZZ006，2021ZZ001）
通讯作者: 黄萍     E-mail: huangping@hmc.edu.cn
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张轶雯
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邹小舟
刘宇佳
黄萍

引用本文:

张轶雯,盛孔胜,宋飞凤,潘宗富,邹小舟,刘宇佳,黄萍. 清肺口服液在特发性肺纤维化中的治疗作用及网络药理学研究[J]. 浙江大学学报（医学版）, 2022, 51(1): 53-61.

ZHANG Yiwen,SHENG Kongsheng,SONG Feifeng,PAN Zongfu,ZOU Xiaozhou,LIU Yujia,HUANG Ping. Efficacy of Qingfei oral liquid for idiopathic pulmonary fibrosis in rats and related network pharmacology study. J Zhejiang Univ (Med Sci), 2022, 51(1): 53-61.

链接本文:

https://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2021-0203    或    https://www.zjujournals.com/med/CN/Y2022/V51/I1/53

图 1  造模第21天各组肺组织形态及纤维化表现（苏木精–伊红染色）A： 正常对照组肺组织结构及纤毛结构正常，肺内细支气管上皮完整，排列整齐，气管和血管周围无明显炎症反应，未见纤维化； B： 模型对照组肺内细支气管上皮增生，管壁增厚，管腔狭窄，伴有炎症细胞浸润，肺泡间隔明显增宽，肺组织均见不同程度的纤维化； C： 吡非尼酮组肺纤维化程度明显缓解，病变程度减轻； D： 清肺口服液组肺纤维化程度与吡非尼酮组相近. 标尺=50 μm.
图 2  造模第21天各组大鼠肺组织中胶原沉积情况（马森染色）A： 正常对照组肺组织中较少蓝色的胶原蛋白沉积； B： 模型对照组肺组织中胶原蛋白沉积增加； C：吡非尼酮组肺组织中胶原蛋白沉积较模型对照组减少； D： 清肺口服液组肺组织中胶原蛋白沉积情况与吡非尼酮组相近. 标尺=10 μm.

　组别

第一周

第二周

第三周

–

±

+

++及以上

–

±

+

++及以上

–

±

+

++及以上

正常对照组

6

0

0

0

6

0

0

0

6

0

0

0

模型对照组

1

1

3

1

0

0

2

4

0

0

1

5

吡非尼酮组

3

1

2

0

0

0

5

1

2

0

3

1

清肺口服液组

1

3

1

1

1

0

0

5

0

0

6

0

P值

>0.05

>0.05

<0.01
表 1  各组肺组织胶原蛋白沉积程度分级分布比较
图 3  各组肺组织中转化生长因子-β（TGF-β）表达A：正常对照组肺组织中TGF-β（棕色） 几乎不表达； B： 模型对照组肺组织中TGF-β表达增加； C： 吡非尼酮组肺组织中TGF-β表达较模型对照组减少； D： 清肺口服液组肺组织中TGF-β表达与吡非尼酮组相近. 标尺=50 μm.
图 4  各组肺组织中GSH、SOD和MDA含量比较<0.05，<0.01. GSH：谷胱苷肽；SOD：超氧化物歧化酶；MDA：丙二醛.
图 5  清肺口服液抗特发性肺纤维化的成分–靶点–疾病网络
图 6  清肺口服液抗特发性肺纤维化潜在靶点的蛋白–蛋白相互作用网络
图 7  清肺口服液抗特发性肺纤维化的作用靶点基因功能注释A： GO富集分析结果排序前十位生物学过程的气泡图； B： GO富集分析结果排序前十位细胞组成的气泡图； C： GO富集分析结果排序前十位分子功能的气泡图.GO：基因本体.
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