运脾泻肺化痰汤对哮喘大鼠肺组织相关蛋白表达的影响
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作者单位:

1.贵州中医药大学第二附属医院 贵州 贵阳550003;2.广元市昭化区妇幼保健院 四川 广元 628021;3.黔西南州中医院 贵州 黔西562400;4.贵州中医药大学 贵州 贵阳 550025

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陈竹,E-mail:918236355@qq.com

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R562.25

基金项目:

贵州省科技厅基础研究计划(No:黔科和基础[2020]1Y365);国家自然科学基金地区科学基金项目(No:81860868);贵州省“十四五”中医药、民族医药重点建设学科(No:黔中医药函[2021])95号-6);贵州省中医儿科诊疗中心建设项目(No:黔中医药函〔2021〕58号


Effects of Yunpi Xiefei Huatan Decoction on expressions of proteins associated with nuclear factor-κB in lung tissues of asthmatic rats
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Affiliation:

1.The Second Affiliated Hospital of Guizhou University of Chinese Medicine, Guiyang, Guizhou 550003, China;2.Zhaohua District Maternal and Child Health Hospital of Guangyuan City, Guangyuan, Sichuan 628021, China;3.Qianxinan Zhou TCM Hospital, Qianxi, Guizhou 562400, China;4.Guizhou University of Chinese Medicine, Guiyang, Guizhou 550025, China

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    摘要:

    目的 探讨运脾泻肺化痰汤对哮喘模型大鼠肺组织相关蛋白表达的影响。方法 采用卵白蛋白诱导复制哮喘大鼠模型,将大鼠分为正常组、模型组、运脾泻肺化痰汤高剂量组(0.020 mL/g)、运脾泻肺化痰汤中剂量组(0.010 mL/g)、运脾泻肺化痰汤低剂量组(0.005 mL/g)、西药组(0.010 mL/g)、联合组(中药0.010 mL/g+西药0.010 mL/g),每组各10只,共7组。分别在第1天和第8天腹腔注射1 mL混悬液[含卵白蛋白100 mg + Al(OH)3100 mg/mL]致敏。第15天开始,用1%卵白蛋白激发,隔日1次,共激发7次,第16天开始灌服治疗,连续4周后采集大鼠肺泡灌洗液及肺组织。行瑞氏-吉姆萨染色观察大鼠肺泡灌洗液中白细胞计数及细胞分类情况;Western blotting检测大鼠肺组织Iκκ-αIκκ-β蛋白表达;实时荧光定量酶聚合酶链反应(qRT-PCR)检测大鼠肺组织中Iκκ-αIκκ-β、IκBα、IκBβ mRNA表达。结果 模型组白细胞计数、嗜酸性粒细胞占比、中性粒细胞占比较正常组高,单核细胞占比较正常组低(P <0.05);运脾泻肺化痰汤高、中、低剂量组、西药组、联合组的白细胞计数、嗜酸性粒细胞占比、中性粒细胞占比较模型组低(P <0.05),运脾泻肺化痰汤高、中、低剂量组、联合组的单核细胞占比较模型组高(P <0.05)。模型组Iκκ-α、Iκκ-β mRNA相对表达量较正常组高(P <0.05),运脾泻肺化痰汤高、中、低剂量组、西药组、联合组Iκκ-α、Iκκ-β mRNA相对表达量较模型组低(P <0.05),联合组Iκκ-α、Iκκ-β mRNA相对表达量较西药组低(P <0.05);模型组IκBα、IκBβ mRNA相对表达量较正常组低(P <0.05),运脾泻肺化痰汤高、中、低剂量组、西药组、联合组IκBα、IκBβ mRNA相对表达量较模型组高(P <0.05),联合组IκBα、IκBβ mRNA相对表达量较西药组高(P <0.05)。模型组Iκκ-αIκκ-β蛋白相对表达量较正常组高(P <0.05),运脾泻肺化痰汤高、中、低剂量组、西药组、联合组Iκκ-αIκκ-β蛋白相对表达量较模型组低(P <0.05),联合组Iκκ-αIκκ-β蛋白相对表达量较西药组低(P <0.05)。结论 运脾泻肺化痰汤可减轻肺泡灌洗液炎症细胞水平,通过上调NF-κB的抑制因子IκBα、IκBβ基因表达,下调Iκκ-α、Iκκ-β蛋白及基因表达量,有减轻哮喘大鼠气道炎症的作用。

    Abstract:

    Objective To investigate the effects of Yunpi Xiefei Huatan Decoction on expressions of proteins associated with nuclear factor-kappa (NF-κB) in lung tissues of asthmatic rats.Methods The rat models of asthma were induced by ovalbumin (OVA) treatment. Specifically, the rats were randomly divided into normal group, model group, high-dose (0.020 mL/g), medium-dose (0.010 mL/g) and low-dose (0.005 mL/g) Yunpi Xiefei Huatan Decoction groups, Western medicine group (0.01 mL/g), and combination group (0.01 mL/g of Chinese medicine + 0.01 mL/g of Western medicine), with 10 rats in each group. The rats were sensitized with intraperitoneal injection of 1 mL of suspension containing 100 mg of ovalbumin and 3100 mg/mL of Al (OH) on day 1 and day 8, followed by stimulation with 1% OVA once every other day since day 15 for a total of 7 sessions. The rats were administrated with according medicines from the 16th day on for a consecutive of 4 weeks, and then the alveolar lavage fluid and lung tissues of rats were collected. The white blood cell (WBC) count and the number of WBC subsets in rat alveolar lavage fluid were determined by Wright-Giemsa staining. Western blotting was used to detect the relative protein expression levels of Iκκ-α and Iκκ-β in rat lung tissues. The mRNA expression levels of Iκκ-α, Iκκ-β, IκBα and IκBβ in rat lung tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR).Results The WBC count and proportions of eosinophils and neutrophils were higher, and the proportion of monocytes was lower in the model group compared with the normal group (P < 0.05). The WBC count and proportions of eosinophils and neutrophils were lower, and the proportion of monocytes was higher in the high-dose, medium-dose and low-dose Yunpi Xiefei Huatan Decoction groups, Western medicine group, and combination group compared with the model group (P < 0.05). The relative mRNA expression levels of Iκκ-α and Iκκ-β were higher in the model group than those in the normal group (P < 0.05), while those in the high-dose, medium-dose and low-dose Yunpi Xiefei Huatan Decoction groups, Western medicine group, and combination group were lower relative to those in the model group (P < 0.05). Besides, the relative mRNA expression levels of Iκκ-α and Iκκ-β in the combination group were lower than those in the Western medicine group (P < 0.05). The relative mRNA expression levels of IκBα and IκBβ in the model group were lower than those in the normal group (P < 0.05), while they were higher in the high-dose, medium-dose and low-dose Yunpi Xiefei Huatan Decoction groups, Western medicine group, and combination group compared with the model group (P < 0.05). The relative mRNA expression levels of IκBα and IκBβ in the combination group were even higher than those in the Western medicine group (P < 0.05). The relative protein expression levels of Iκκ-α and Iκκ-β in the model groups were higher than those in the normal group (P < 0.05), whereas they were lower in the high-dose, medium-dose and low-dose Yunpi Xiefei Huatan Decoction groups, Western medicine group, and combination group compared with the model group (P < 0.05). In addition, the relative protein expression levels of Iκκ-α and Iκκ-β in the combination group were even lower than those in the Western medicine group (P < 0.05).Conclusions Yunpi Xiefei Huatan Decoction may reduce the levels of pro-inflammatory cells in the alveolar lavage fluid, and upregulate the gene expressions of NF-κB inhibitors IκBα and IκBβ while downregulating the expressions of Iκκ-α and Iκκ-β, thereby alleviating the airway inflammation in asthmatic rats.

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陈娜,陈竹,刘志伟,杜丽,李艳静,阳和平,骆弟琴,王文洲,曹正连.运脾泻肺化痰汤对哮喘大鼠肺组织相关蛋白表达的影响[J].中国现代医学杂志,2023,(14):9-15

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  • 收稿日期:2022-09-06
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  • 在线发布日期: 2023-12-04
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