摘要
目的探讨艾蒿花粉中脂质和脂多糖(lipopolysaccharide,LPS)对诱导季节性变应性鼻炎小鼠模型效果的影响。方法采用随机数字表法将BALB/c小鼠随机分为7组,分别为脱脂花粉组(TZ组,n=5)、脱脂花粉+LPS组(TZLPS组,n=5)、未脱脂花粉组(WTZ组,n=5)、未脱脂花粉+LPS组(WTZLPS组,n=5)、PBS组(n=5)、PBS+LPS组(PBSLPS组,n=5)、空白对照组(n=6)。在第1、8、15天,TZ、TZLPS组小鼠颈部皮下注射0.1 ml的脱脂艾蒿花粉提取液(20μg/ml),WTZ、WTZLPS组给予0.1 ml的未脱脂艾蒿花粉提取液(20μg/ml),PBS、PBSLPS组给予0.1 ml的PBS溶液(0.1 mol/L);在第22~28天,TZ组、WTZ组、PBS组分别用脱脂花粉提取液500μg/ml、未脱脂花粉提取液500μg/ml、PBS溶液0.1 mol/L滴鼻激发,每只小鼠每鼻孔10μl,TZLPS、WTZLPS、PBSLPS组在以上基础上每只小鼠每鼻孔分别加260 EU(5.2μl)LPS。空白对照组不进行任何干预。观察小鼠行为症状改变,ELISA法检测小鼠血清中特异性IgE、IgG1、IgG2a水平,HE染色法观察鼻黏膜和肺组织病理变化,免疫组化法观察IL-4、IL-5表达情况。统计学分析采用单因素方差分析和Kruskal-Wallis检验。结果经艾蒿花粉抗原激发后,总TZ组(TZLPS组+TZ组)小鼠的行为学评分显著高于总PBS组(PBSLPS组+PBS组)和空白对照组(P均<0.001);总TZ组小鼠血清特异性IgE水平较总WTZ组(WTZLPS组+WTZ组)和总PBS组显著升高(P<0.05,P<0.001),其IgG1水平显著高于总PBS组和空白对照组(P<0.05,P<0.001),而IgG2a水平与总PBS组和空白对照组的差异无统计学意义(P均>0.05)。在局部炎症反应中,总TZ组鼻黏膜和肺组织中嗜酸性粒细胞计数以及IL-4、IL-5表达均显著高于总PBS组和空白对照组(P均<0.001);且鼻组织嗜酸性粒细胞计数显著高于总WTZ组(P<0.05),但总TZ组与总WTZ组间鼻黏膜IL-4、IL-5表达水平差异无统计学意义(P均>0.05)。花粉提取液中是否含有LPS对各组小鼠血清中特异性IgE、IgG1和IgG2a水平影响均无显著差异(P均>0.05),对鼻黏膜中嗜酸性粒细胞活化和IL-4的阳性表达影响也无明显差异(P均>0.05);但在肺组织中,TZLPS组嗜酸性粒细胞计数和IL-4表达要显著高于TZ组(P均<0.001)。结论本研究成功建立艾蒿花粉变应原过敏小鼠模型,并发现对艾蒿花粉进行脱脂处理能引发更强烈的炎症反应,花粉中是否含有LPS对鼻黏膜局部过敏性炎症影响不显著,但对小鼠肺组织造成不同程度损伤。
Objective To investigate the effects of lipids and lipopolysaccharide(LPS)in Artemisia pollen on the induction of allergic rhinitis in a murine model.Methods BALB/c mice were randomly divided into seven groups using the random number table method as follows:defatted pollen group(TZ group,n=5),defatted pollen+LPS group(TZLPS group,n=5),non-defatted pollen group(WTZ group,n=5),non-defatted pollen+LPS group(WTZLPS group,n=5),PBS group(n=5),PBS+LPS group(PBSLPS group,n=5),and blank control group(n=6).On days 1,8,and 15,the mice in the TZ and TZLPS groups received a subcutaneous injection in the neck region with 0.1 ml of defatted Artemisia pollen extract(20μg/ml);the WTZ and WTZLPS groups were administered 0.1 ml of non-defatted Artemisia pollen extract(20μg/ml),while the PBS and PBSLPS groups were injected with 0.1 ml of PBS(0.1 mol/L).From days 22 to 28,the mice were subjected to intranasal challenge to induce allergic rhinitis symptoms.The TZ,WTZ,and PBS groups received nasal administration of 10μl per nostril of defatted Artemisia pollen extract(500μg/ml),non-defatted Artemisia pollen extract(500μg/ml),and PBS(0.1 mol/L),respectively.For the TZLPS,WTZLPS,and PBSLPS groups,an additional 260 EU(5.2μl)of LPS was co-administered per nostril alongside the corresponding base solutions.The blank control group received no intervention during this phase.The behaviors of the mice were observed;the levels of specific IgE,IgG1 and IgG2a in serum samples were detected by ELISA;the pathological changes in nasal mucosa and lung tissues were observed by HE staining,and the expression of both IL-4 and IL-5 was observed by immunohistochemistry.One-way analysis of variance and Kruskal-Wallis test were used for statistical analysis.Results Following Artemisia pollen antigen challenge,the total TZ group(TZLPS group+TZ group)exhibited significantly higher behavioral scores as compared with the total PBS group(PBSLPS group+PBS group)and the blank control group(both P<0.001).Serum analysis revealed that the total TZ group showed markedly elevated levels of Artemisia-specific IgE compared with the total WTZ group(WTZLPS group+WTZ group)and the total PBS group(P<0.05,P<0.001),along with significantly higher IgG1 levels than the total PBS and blank control groups(P<0.05,P<0.001),while no significant differences in IgG2a levels were observed among these groups(all P>0.05).In local inflammatory responses,eosinophil infiltration and IL-4/IL-5 expression in both nasal mucosa and lung tissues of mice in the total TZ group were significantly higher than those in the total PBS and blank control groups(all P<0.001).Notably,eosinophil counts in nasal mucosa of mice in the total TZ group surpassed those in the total WTZ group(P<0.05),whereas no significant differences in IL-4/IL-5 expression in mouse nasal mucosa were detected between the total TZ and total WTZ groups(both P>0.05).LPS supplementation in pollen extracts showed no significant effects on the specific IgE,IgG1,or IgG2a levels in serum across groups(all P>0.05),nor did it alter eosinophil activation or IL-4 expression in mouse nasal mucosa(all P>0.05).However,compared with the TZ group,eosinophil counts and IL-4 expression in lung tissues of mice in the TZLPS group were significantly increased(both P<0.001).Conclusions This study successfully establishes a mouse model of Artemisia pollen allergy,and finds that the defatting treatment of Artemisia pollen can induce more intense inflammatory response.The presence or absence of LPS in pollen has no significant effect on allergic inflammation in the nasal mucosa,but it can cause different degrees of damage to the lung tissues of mice.
作者
徐丹
高斐宏
李姗姗
王毅力
Xu Dan;Gao Feihong;Li Shanshan;Wang Yili(College of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《中华微生物学和免疫学杂志》
北大核心
2025年第5期378-386,共9页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(8210153109)。
关键词
变应性鼻炎
艾蒿花粉
动物模型
脂多糖
脂质
Allergic rhinitis
Artemisia pollen
Animal model
Lipopolysaccharide
Lipids
