通鼻消涕颗粒治疗大鼠慢性鼻窦炎的作用机制研究
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云南中医药大学第一附属医院

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本课题获国家自然科学基金(编号:81960890)及云南省科技厅中医联合专项课题基金资助(编号:2018FF001-034)


Study on the mechanism of Tongbixiaoti Granules in the treatment of chronic rhinosinusitis in rats
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The department of Otorhinolaryngology,First Affiliated Hospital of Yunnan Universitiy of Traditional Chinese Medicine

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    摘要:

    目的:本课题通过动物实验研究为通鼻消涕颗粒治疗慢性鼻窦炎提供客观实验数据支持。方法:SPF级SD大鼠60只,取10只大鼠作为正常对照组,其余随机分为模型组、克拉霉素片组、通鼻消涕颗粒低、中、高剂量组,每组10只。造模成功后分别应用克拉霉素和通鼻消涕颗粒灌胃治疗2周后,HE染色检测鼻粘膜炎症情况。免疫组化染色及RT-PCR实验检测各组鼻粘膜组织的PLUNC、Cu/ZnSOD、PACAP、DJ-1、谷胱苷肽S转移酶-pi、NKEF-B的蛋白和mRNA。ELISA法检测大鼠血清的NF-κB表达水平。流式细胞术检测大鼠外周血Th淋巴细胞亚群Th1/Th2水平。采用t检验进行统计分析,对比服药前、服药后各个阶段的免疫指标含量变化有无显著差异。结果:HE染色观察大鼠鼻粘膜组织病理改变情况及免疫组化染色发现PLUNC等鼻粘膜保护性蛋白均表达在鼻粘膜纤毛柱状上皮及粘膜下腺体中。RT-PCR检测结果表明经通鼻消涕颗粒治疗后,大鼠鼻粘膜上皮及粘膜下腺体反应性增生,产生PLUNC等鼻粘膜保护性蛋白增多来抵抗慢性炎症,克拉霉素治疗后主要是促进炎症消退,而鼻粘膜保护性蛋白产生不多。ELISA检测大鼠血清的NF-κB表达水平提示:通鼻消涕颗粒及克拉霉素片均能减轻慢性鼻窦炎大鼠血清中的NF- κ B水平达到抗炎效果。流式检测结果表明中药通鼻消涕颗粒和克拉霉素片均能提高Th1细胞比例,降低Th2细胞比例,从而有助于促进炎症消退。结论:中药通鼻消涕颗粒可通过提高鼻粘膜组织中PLUNC等鼻粘膜保护性蛋白含量来改善鼻粘膜局部免疫功能并降低血清NF-κB和Th2细胞比例促进炎症消退从而达到“扶正祛邪”的治疗效果。

    Abstract:

    Objective: This study provides objective experimental data support for the treatment of chronic sinusitis (CRS) with Tongbi Xiaoti granules through animal experiments. Method: 60 SPF grade SD rats were selected, with 10 rats as the normal control group. The rest were randomly divided into model group, clarithromycin tablet group, low, medium, and high dose groups of Tongbi Xiaoti granules, with 10 rats in each group. After successful modeling, clarithromycin and Tongbi Xiaoti granules were administered by gavage for 2 weeks, and HE staining was used to detect nasal mucositis. Immunohistochemical staining and RT-PCR assay were used to detect PLUNC、Cu/ZnSOD、PACAP、DJ-1、glutathione S-transferase pi and NKEF-B . ELISA method was used to detect the expression level of NF - κ B in rat serum. Flow cytometry was used to detect the Th1/Th2 levels of peripheral blood Th lymphocyte subsets in rats. Use t-test for statistical analysis to compare whether there is a significant difference in the changes in immune index content before and after medication at each stage. Result: HE staining was used to observe the pathological changes of rat nasal mucosa tissue, and immunohistochemical staining revealed that protective proteins such as PLUNC were expressed in the ciliated columnar epithelium and submucosal glands of the nasal mucosa. The RT-PCR test results showed that after treatment with Tongbi Xiaoti Granules, the epithelial and submucosal glands of the rat nasal mucosa showed reactive proliferation, producing more protective proteins such as PLUNC to resist chronic inflammation. Clarithromycin treatment mainly promoted inflammation resolution, while the production of protective proteins in the nasal mucosa was not significant. ELISA detection of NF - κ B expression levels in rat serum suggests that both Tongbi Xiaoti Granules and Clarithromycin Tablets can reduce NF - κ B levels in the serum of rats with CRS and achieve anti-inflammatory effects. The flow cytometry results showed that both Tongbi Xiaoti granules and clarithromycin tablets can increase the proportion of Th1 cells and reduce the proportion of Th2 cells, thereby helping to promote inflammation resolution. Conclusion: Traditional Chinese medicine Tongbi Xiaoti Granules can improve local immune function of the nasal mucosa and reduce the ratio of serum NF - κ B and Th2 cells by increasing the content of protective proteins such as PLUNC in the nasal mucosa tissue, promoting inflammation resolution and achieving the therapeutic effect of "strengthening the body resistance to eliminate pathogenic factors".

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  • 收稿日期:2024-04-03
  • 最后修改日期:2024-06-07
  • 录用日期:2024-06-11
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