1.湖南中医药大学第一附属医院,湖南省长沙市雨花区韶山中路95号,410007
2.深圳市前海蛇口自贸区医院
3.湖南中医药大学
wmqwmq2009@sina. com,(0731)85369185
收稿:2022-05-20,
修回:2022-07-18,
纸质出版:2022-12-17
移动端阅览
罗菁,丁伊,詹国平等.感染后咳嗽肺热阴虚证幼年大鼠模型的构建及验证[J].中医杂志,2022,63(24):2366-2374.
LUO Jing,DING Yi,ZHAN Guoping,et al.Construction and Verification of Post-infectious Cough Model with Lung Heat and Yin Deficiency Syndrome in Juvenile Rats[J].Journal of Traditional Chinese Medicine,2022,63(24):2366-2374.
罗菁,丁伊,詹国平等.感染后咳嗽肺热阴虚证幼年大鼠模型的构建及验证[J].中医杂志,2022,63(24):2366-2374. DOI: 10.13288/j.11-2166/r.2022.24.012.
LUO Jing,DING Yi,ZHAN Guoping,et al.Construction and Verification of Post-infectious Cough Model with Lung Heat and Yin Deficiency Syndrome in Juvenile Rats[J].Journal of Traditional Chinese Medicine,2022,63(24):2366-2374. DOI: 10.13288/j.11-2166/r.2022.24.012.
目的
2
探讨感染后咳嗽肺热阴虚证幼年大鼠模型的建立与验证。
方法
2
40只幼年SD大鼠随机分为空白组、模型组、泻白增液汤组、橘红痰咳液组,每组10只。除空白组外,其余各组大鼠采用烟熏+脂多糖滴鼻+辣椒素雾化制备感染后咳嗽幼年大鼠模型。实验第1~18天为造模阶段,实验第19~26天为以方测证阶段。实验第19天开始泻白增液汤组给予浓度为1.48 g/ml的泻白增液汤水煎液10 ml/kg灌胃,橘红痰咳液组给予橘红痰咳液5 ml/kg灌胃,空白组、模型组大鼠给予生理盐水10 ml/kg灌胃。各组均每日灌胃1次,持续7天。分别于实验第1、10、19天,比较空白组和模型组大鼠肛温、四肢爪心温度、心前区温度、体质量、粪便含水量、饮水量变化。实验第26天,比较四组大鼠肛温、四肢爪心温度、心前区温度、体质量、粪便含水量、饮水量变化。实验第27天进行咳嗽敏感性测定,记录大鼠咳嗽潜伏期和5 min内咳嗽次数;测定肺功能指标[包括呼气时间(TE)、增强呼气间歇(PENH)、呼气末暂停时间(EEP)、松弛时间(RT)];检测肺泡灌洗液中细胞分类计数及炎性因子[包括肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)]水平;检测血清环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)含量,并计算cAMP/cGMP值;HE染色观察肺组织病理。
结果
2
实验第19天,与空白组比较,模型组大鼠体质量降低,粪便含水量减少,饮水量增加,大鼠肛温、四肢爪心温度、心前区温度升高(
P
<0.05或
P
<0.01)。实验第26天,与空白组比较,模型组大鼠体质量、粪便含水量下降,肛温和四肢爪心温度升高,饮水量增加,大鼠咳嗽潜伏期、TE、EEP明显缩短,5 min内咳嗽次数、RT、PENH明显增加,肺泡灌洗液中中性粒细胞、巨噬细胞、淋巴细胞、嗜酸性粒细胞计数及IL-1β、TNF-α水平升高,血清cAMP、cAMP/cGMP亦升高(
P
<0.05或
P
<0.01)。与模型组比较,泻白增液汤组大鼠体质量和粪便含水量增加,肛温和四肢爪心温度降低,饮水量减少,咳嗽潜伏期、TE、EEP均明显延长,RT、PENH降低,5min内咳嗽次数均减少,肺泡灌洗液中中性粒细胞、巨噬细胞、淋巴细胞、嗜酸性粒细胞计数及IL-1β、TNF-α水平降低,血清cAMP、cAMP/cGMP均降低(
P
<0.05);橘红痰咳液组仅咳嗽潜伏期、TE、EEP均延长,5 min内咳嗽次数减少,中性粒细胞、淋巴细胞计数及TNF-α水平降低(
P
<0.05)。HE染色结果显示,空白组大鼠肺及支气管组织基本正常;模型组肺组织多见血管充血、淤血,支气管及支气管周围炎性改变,较大面积的肺泡壁中度增厚,肺泡间距增宽;泻白增液汤组、橘红痰咳液组较模型组肺组织病理损伤均有不同程度减轻,两组之间对比无明显差异。
结论
2
采用联合烟熏+脂多糖滴鼻+辣椒素雾化制备的感染后咳嗽幼年大鼠模型,具备肺热阴虚证候特征。
Objective
2
To investigate the establishment of post-infectious cough model with lung heat and yin deficiency syndrome in juvenile rats, and to verify the model.
Methods
2
Forty juvenile SD rats were randomly divided into blank group, model group,
Xiebai Zengye Decoction
(泻白增液汤) group and
Juhong Tanke Liquid
(橘红痰咳液) group, with 10 rats in each group. Except for the blank group, rats were exposed to smoke and given lipopolysaccharide nasal drops and capsaicin atomization to prepare post-infectious cough juvenile rat model. The 1st to 18th day was the modeling stage, and the 19th to 26th day of the experiment was the verification stage of the formulas. Since the 19th day, 10 ml/kg of
Xiebai Zengye Decoction
with a concentration of 1.48 g/ml and 5 ml/kg of
Juhong Tanke Liquid
were given by gavage to the corresponding two groups, respectively, while 10 ml/kg of normal saline was administered by gavage to the blank group and model group, once daily for seven days. On the 1st, 10th, and 19th day of the experiment, the rats in the blank group and the model group were compared for changes in anal temperature, paw temperature, precordial temperature, body weight, feces moisture content, and water intake. On the 26th day, the outcomes mentioned above were measured and compared among the four groups. On the 27th day, the cough sensitivity was measured, and the cough latency and the number of coughs within 5 minutes were recorded. The pulmonary function indicators including expiratory time (TE), enhanced expiratory pause (PENH), end-expiratory pause (EEP) and relaxation time (RT) were detected, as well as the alveolar lavage fluid cell differential counts and inflammatory factors including tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β)] levels. The content of serum cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were detected, and the value of cAMP/cGMP was calculated. Lung histopathology was observed by HE staining.
Results
2
On the 19th day of the experiment, compared to those in the blank group, the body weight of the rats and the feces moisture content significantly decreased in the model group, while the water intake, anal temperature, paw temperature and precordial temperature increased (
P
<0.05 or
P
<0.01). On the 26th day, the model group had significantly decreased body weight and feces moisture content, increased anal and paw temperatures, increased water intake, shortened cough latency, TE and EEP, growed cough number within 5 minutes, RT and PENH, increased levels of neutrophils, macrophages, lymphocytes, eosinophils in the alveolar lavage fluid, as well as the IL-1β and TNF-αlevels, and serum cAMP level and cAMP/cGMP value than the blank group (
P
<0.05 or
P
<0.01). Compared to those in the model group, the body weight and feces moisture content in the
Xiebai Zengye Decoction
group increased, with decreased anal and paw temperatures, reduced water intake, prolonged cough latency, TE and EEP, decreased RT, PENH and cough number within 5 minutes, reduced levels of neutrophils, macrophages, lymphocytes, eosinophils in the alveolar lavage fluid, as well as the IL-1β and TNF-αlevels, and serum cAMP level and cAMP/cGMP value (
P
<0.05); the cough latency, TE and EEP were prolonged in the
Juhong Tanke Liquid
group, with reduced cough number within 5 minutes, decreased neutrophils and lymphocytes, and the level of TNF-α (
P
<0.05). HE staining showed that the lung and bronchi tissues of the rats in the blank group were basically normal; the lung tissues of the model group were mostly seen with vascular hyperemia and congestion, inflammatory changes in and around the bronchi, large area of moderately thickened alveolar walls, and widened alveolar spacing; the pathological damage of lung tissues in the
Xiebai Zengye Decoction
group and
Juhong Tanke Liquid
group were alleviated to varying degrees than in the model group, but without significant differences.
Conclusion
2
The post-infectious cough juvenile rat model prepared by combination of smoke exposure, lipopolysaccharide nasal drops and capsaicin atomization has the characteristics of lung heat and yin deficiency syndrome.
洪建国 . 《中国儿童咳嗽诊断与治疗临床实践指南(2021年版)》解读 [J]. 中华医学信息导报 , 2021 , 36 ( 22 ): 21 .
河智研 . 基于数据挖掘的肺炎支原体感染后儿童慢性咳嗽证治规律分析 [D]. 沈阳 : 辽宁中医药大学 , 2017 .
朱梦婷 . 小儿感染后咳嗽中医证型的归类分析 [D]. 乌鲁木齐 : 新疆医科大学 , 2016 .
林传权 . 病证结合动物模型研究的进展及存在问题分析 [J]. 中华中医药杂志 , 2019 , 34 ( 8 ): 3601 - 3604 .
李飞侠 , 朱佳 , 王文 , 等 . 一种感染后咳嗽动物模型制作方法的建立及评价 [J]. 中华中医药学刊 , 2013 , 31 ( 10 ): 2315 - 2317 .
王赛 , 白明 , 苗明三 . 阴虚证动物模型诊断指标及分析 [J]. 中国比较医学杂志 , 2021 , 31 ( 1 ): 132 - 137 .
周婷 , 王孟清 , 罗银河 . 肺热阴虚型感染后慢性咳嗽大鼠模型的建立及评价 [J]. 中医药导报 , 2016 , 22 ( 5 ): 16 - 20 .
宋亚刚 , 李艳 , 崔琳琳 , 等 . 中医药病证结合动物模型的现代应用研究及思考 [J]. 中草药 , 2019 , 50 ( 16 ): 3971 - 3978 .
钱乙 . 小儿药证直诀 [M]. 郭君双,整理.北京 : 人民卫生出版社 , 2006 : 76 .
吴瑭 . 温病条辨 [M]. 南京中医药大学温病学教研室,整理.北京 : 人民卫生出版社 , 2005 : 105 .
徐叔云 , 卞如濂 , 陈修 . 药理实验方法学 [M]. 北京 : 人民卫生出版社 , 1982 : 305 .
彭丹虹 , 王燕萍 , 刘晓琪 , 等 . 中医病证结合动物模型评价体系的现状分析 [J]. 中华中医药学刊 , 2017 , 35 ( 8 ): 2027 - 2030 .
任钧国 , 刘建勋 , 韦佳慧 . 病证结合药理学:中药药理学研究的策略与方法 [J]. 中国中药杂志 , 2021 , 46 ( 19 ): 4922 - 4929 .
辛伟 . 基于下丘脑-垂体-性腺轴季节性功能变化的绝经期大鼠“潮热”的生物学机制研究 [D]. 晋中 : 山西中医药大学 , 2020 .
孙晓霞 , 任威铭 , 崔荣兴 , 等 . 阴虚证动物模型的建立与评价 [J]. 中医杂志 , 2021 , 62 ( 13 ): 1156 - 1163 .
翟建宾 , 赵亮 , 于士昌 , 等 . 桑苏饮调控TLR4通路抑制哮喘大鼠气道炎症 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 14 ): 21 - 28 .
TAITANO SH , VAN DER VLUGT L , SHEA MM , et al . Differential influence on regulatory B cells by TH2 cytokines affects protection in allergic airway disease [J]. J Immunol , 2018 , 201 ( 7 ): 1865 - 1874 .
BARNIG C , LEVY BD . Innate immunity is a key factor for the resolution of inflammation in asthma [J]. Eur Respir Rev , 2015 , 24 ( 135 ): 141 - 153 .
王志旺 , 李荣科 , 刘雪枫 , 等 . 当归对阴虚哮喘小鼠模型的影响 [J]. 中国实验方剂学杂志 , 2016 , 22 ( 21 ): 116 - 120 .
陈进成 , 刘建勋 , 马博 , 等 . 气虚血瘀证大鼠模型的建立与评价 [J]. 中医杂志 , 2020 , 61 ( 3 ): 229 - 234 .
0
浏览量
5
下载量
4
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010102007558号