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彭果然, 程贝贝, 丁蓉珍, 戴爱国.哮喘肝郁证病证结合小鼠模型的建立与评价[J].湖南中医药大学学报,2025,45(2):204-211[点击复制] |
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| 哮喘肝郁证病证结合小鼠模型的建立与评价 |
| 彭果然,程贝贝,丁蓉珍,戴爱国 |
| (湖南中医药大学中西医结合学院, 湖南 长沙 410208;血管生物学与转化医学湖南省重点实验室, 湖南 长沙 410208;湖南中医药大学医学院呼吸疾病研究室, 湖南 长沙 410208;湖南中医药大学第一附属医院呼吸内科, 湖南 长沙 410007) |
| 摘要: |
| 目的 建立C57BL/6小鼠哮喘肝郁证病证结合模型并进行评价。方法 50只C57BL/6小鼠随机分为空白组、哮喘组、哮喘肝郁组、布地奈德组、柴朴汤组。除空白组外，其余4组小鼠均采用屋尘螨（HDM）25 μg滴鼻，1次/d，5 d/周，持续4周进行哮喘造模，其中哮喘肝郁组、布地奈德组和柴朴汤组同期联合予以慢性不可预知性温和刺激（CUMS），1次/d，持续28 d制备哮喘肝郁证模型。布地奈德组（吸入用布地奈德混悬液0.5 g/L雾化吸入）和柴朴汤组（柴朴汤12.48 g/kg灌胃）于造模第15～28天接受治疗。观察各组小鼠一般状况、体质量，并进行糖水偏好实验、强迫游泳实验，检测肺功能及血清促肾上腺皮质激素（ACTH）、白细胞介素-6（IL-6）水平，HE染色及Masson染色观察肺组织病理情况。结果 第28天实验结束后，与空白组比较，哮喘组与哮喘肝郁组一般状况评分、肺阻力（RL）、ACTH及IL-6均显著升高（P<0.01），用力肺活量（FVC）及呼气峰流量（PEF）均明显减少（P<0.01），HE染色及Masson染色示气道周围炎症细胞浸润及胶原纤维沉积明显；哮喘肝郁组体质量、糖水偏好指数明显下降（P<0.01），游泳不动时间明显增加（P<0.01）。与哮喘组比较，哮喘肝郁组体质量、糖水偏好指数、FVC及PEF均明显下降（P<0.01），游泳不动时间、ACTH及IL-6均显著升高（P<0.01），HE染色及Masson染色示气道周围炎症细胞浸润及胶原纤维沉积更严重。与哮喘肝郁组比较，布地奈德组及柴朴汤组一般状况评分、游泳不动时间、RL及IL-6均明显下降（P<0.01），体质量、FVC及PEF均明显升高（P<0.01），HE染色及Masson染色示气道周围炎症细胞浸润及胶原纤维沉积减轻；柴朴汤组糖水偏好指数明显升高（P<0.01），ACTH明显下降（P<0.01）。与布地奈德组比较，柴朴汤组体质量及糖水偏好指数均升高（P<0.05），游泳不动时间减少（P<0.05），HE染色及Masson染色示气道周围炎症细胞浸润及胶原纤维沉积程度更轻。结论 采用HDM 25 μg滴鼻，1次/d，5 d/周，持续4周，同时联用28 d CUMS能够成功构建小鼠哮喘肝郁证病证结合模型。 |
| 关键词: 哮喘 肝郁证 动物模型 病证结合 柴朴汤 |
| DOI：10.3969/j.issn.1674-070X.2025.02.003 |
| 投稿时间：2024-10-29 |
| 基金项目:湖南省教育厅科学研究项目（22B0383）；湖南省研究生科研创新项目（CX20230819）；湖南中医药大学校级研究生创新课题（2023CX117）；湖南中医药大学-湖南明康中锦医疗科技发展有限公司联合基金项目（2022MKZJ08）；长沙市自然科学基金项目（kq2208184）。 |
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| Establishment and evaluation of a mouse model combining asthma with liver qi stagnation pattern |
| PENG Guoran, CHENG Beibei, DING Rongzhen, DAI Aiguo |
| (School of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Key Laboratory of Vascular Biology and Translational Medicine, Education Department of Hunan Province, Changsha, Hunan 410208, China;Research Laboratory of Respiratory Diseases, School of Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Respiratory Medicine Department, the First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To establish and evaluate a model combining asthma with liver qi stagnation pattern in C57BL/6 mice. Methods Fifty C57BL/6 mice were randomized into blank group, asthma group, asthma with liver qi stagnation group, budesonide group, and Chaipo Decoction (CPD) group. Except for the blank group, the other four groups of mice were intranasally administered with 25μg of house dust mite (HDM) once a day, five days a week, for four weeks to induce the asthma model. Among them, the asthma with liver qi stagnation group, budesonide group, and CPD group were simultaneously subjected to chronic unpredictable mild stress (CUMS) once a day for 28 days to establish the asthma with liver qi stagnation pattern model. The budesonide group (receiving nebulized inhalation of budesonide suspension at 0.5 g/L) and the CPD group (receiving gastric gavage of CPD at 12.48 g/kg) were treated from day 15 to day 28 of modeling. The general condition and body weight of mice in each group were observed, sucrose preference test and forced swimming test were conducted, and lung function and serum levels of adrenocorticotropic hormone (ACTH) and interleukin-6 (IL-6) were measured. Additionally, pathological conditions of lung tissue were observed via HE staining and Masson staining. Results At the end of the 28-day experiment, compared with the blank group, the asthma group and the asthma with liver qi stagnation group showed significant increases in general condition scores, pulmonary resistance (RL), and levels of ACTH and IL-6 (P<0.01), as well as significant decreases in forced vital capacity (FVC) and peak expiratory flow (PEF) (P<0.01). HE staining and Masson staining revealed marked infiltration of inflammatory cells and deposition of collagen fibers around the airways. Additionally, the asthma with liver qi stagnation group exhibited significant decreases in body weight and sucrose preference index (P<0.01), along with a significant increase in immobility time during swimming (P<0.01). Compared with the asthma group, the asthma with liver qi stagnation group showed significant decreases in body weight, sucrose preference index, FVC, and PEF (P<0.01). Furthermore, there were significant increases in immobility time during swimming, ACTH, and IL-6 levels (P<0.01). HE staining and Masson staining revealed more severe inflammatory cell infiltration and collagen fiber deposition around the airways. Compared with the asthma with liver qi stagnation group, the budesonide group and the CPD group showed significant decreases in general condition score, immobility time during swimming, RL, and IL-6 levels (P<0.01), and significant increases in body weight, FVC, and PEF (P<0.01). HE staining and Masson staining revealed reduced inflammatory cell infiltration and collagen fiber deposition around the airways. Furthermore, the CPD group showed a significant increase in sucrose preference index (P<0.01) and a significant decrease in ACTH level (P<0.01). Compared with the budesonide group, the CPD group showed significant increases in body weight and sucrose preference index (P<0.05), as well as a decrease in immobility time during swimming (P<0.05). HE staining and Masson staining revealed milder inflammatory cell infiltration and collagen fiber deposition around the airways. Conclusion A mouse model combining asthma with liver qi stagnation pattern can be successfully established by intranasal administration of HDM 25 μg, once daily, five days a week for four weeks, combined with 28 days of CUMS. |
| Key words: asthma liver qi stagnation pattern animal model combination of disease and pattern Chaipo Decoction |
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