NOD样受体蛋白3信号通路在变应性鼻结膜炎中的作用

doi:10.3969/j.issn.1006-5725.2024.14.004

摘要/Abstract

摘要：

目的建立变应性鼻结膜炎小鼠模型，研究Nod样受体蛋白3（NLRP3）信号通路在变应性鼻结膜炎中的作用。方法 33只雌性C57小鼠（SPF级）随机分为3组：对照组，实验组，NLRP3^-/-组。实验组和NLRP3^-/-组分别于第0、4、7、14、21天行腹腔注射含卵清蛋白（OVA） 100 μg与佐剂Al（OH） 0.2 mL/只。间隔3 d后，每周连续5 d用5% OVA分别点每眼每鼻各10 μL诱发过敏症状；在致敏和激发阶段，对照组均用等量PBS替代。观察小鼠眼部、鼻部症状并进行评分。应用ELISA方法检测血清中OVA特异性IgE、IL-4、IL-17及IL-18含量。HE染色检测小鼠睑结膜及鼻黏膜组织变化。Real-time PCR 检测睑结膜及鼻黏膜组织NLRP3 mRNA的表达。结果实验组和NLRP3^-/-组均诱发出鼻部、眼部过敏症状；实验组鼻部出现过敏症状时间为（10.500 ± 1.080）d，眼部出现过敏症状时间为（20.300 ± 2.058）d；NLRP3^-/-组鼻部出现过敏症状时间为（13.400 ± 1.955）d，眼部出现过敏症状时间为（20.900 ± 2.132）d；NLRP3^-/-组鼻部过敏时间较实验组显著延长（P < 0.05），但眼部过敏时间同实验组差异无统计学意义（P > 0.05）。实验组及NLRP3^-/-组血清OVA特异性IgE及IL-4、IL-17水平均高于对照组，差异均有统计学意义（P < 0.05）。实验组血清IL-18含量较对照组及NLRP3^-/-组显著增加（P < 0.05）。实验组及NLRP3^-/-组睑结膜及鼻黏膜组织病变明显。实验组睑结膜及鼻黏膜NLRP3 mRNA表达较对照组及NLRP3^-/-组显著增加（P < 0.05）。结论变应性鼻结膜炎发病机制复杂，受多因素影响；NLRP3信号通路在其发病中起一定的促进作用。

关键词: 变应性鼻结膜炎, 卵清蛋白, IgE, NLRP3

Abstract:

Objective The objective of this study was to establish a mouse model of allergic rhinoconjunctivitis and investigate the role of the NLRP3 signaling pathway in allergic rhinoconjunctivitis. Methods Thirty-three female C57 mice （SPF） were randomLy divided into 3 groups： the control group， the experimental group， and the NLRP3^-/- group. On days 0， 4， 7， 14， and 21， the experimental group and NLRP3^-/- group received a 0.2 mL intraperitoneal injection of medicine containing OVA （100 μg） and adjuvant Al（OH）₃ （4 mg）， respectively. After an interval of 3 days， each eye and nose were dosed with 10 μL of 5% OVA for five consecutive days a week to induce allergic symptoms. During sensitization and excitation stages， the control group was replaced with an equivalent amount of PBS. Ocular and nasal symptoms were observed and scored. The levels of OVA-specific IgE， IL-4， IL-17， and IL-18 in serum were measured using ELISA， while changes in palpebral conjunctiva and nasal mucosa were assessed by hematoxylin-eosin staining. The expression of NLRP3 mRNA in conjunctival tissue and nasal mucosa was determined using real-time PCR analysis. Statistical analysis was performed using SPSS17.0 software with P < 0.05 considered as statistically significant difference. Results The experimental group and NLRP3^-/- group exhibited induced nasal and ocular allergic symptoms. In the experimental group， the duration of nasal allergy symptoms was （10.500 ± 1.080） days， while the duration of eye allergy symptoms was （20.300 ± 2.058） days. In the NLRP3^-/- group， the duration of nasal allergy symptoms was （13.400 ± 1.955） days， and for eye allergy symptoms it was （20.900 ± 2.132） days. The duration of nasal allergies in the NLRP3^-/- group significantly exceeded that in the experimental group （P < 0.05）， whereas there were no significant differences observed in eye allergy durations between these two groups （P > 0.05）. Levels of OVA-specific IgE， IL-4， and IL-17 were significantly higher in both the experimental and NLRP3^-/- groups compared to those in the control group （P < 0.05）. Additionally， serum IL-18 content increased significantly in the experimental group when compared with both control and NLRP3^-/- groups （P < 0.05）. Conjunctival tissue lesions as well as nasal mucosa damage were evident in both experimental and NLRP3^-/- groups. mRNA expression levels of NLRP3 within conjunctival tissue and nasal mucosa from the experimental group showed a significant increase when compared to those from both control and NLRP3^-/- groups（P < 0.05）. Conclusion Allergic rhinoconjunctivitis pathogenesis is influenced by various factors； however， the involvement of NLPR3 signaling pathway promotes its development.

Key words: allergic rhinoconjunctivitis, ovalbumin, IgE, NLRP3

中图分类号:

R765.2

宫玉波,郭小华,芦文俊,李元超,仇长宇,石圆圆,夏丽萍,石琳,吴玮,罗灵. NOD样受体蛋白3信号通路在变应性鼻结膜炎中的作用[J]. 实用医学杂志, 2024, 40(14): 1922-1927.

Yubo GONG,Xiaohua GUO,Wenjun LU,Yuanchao LI,Changyu QIU,Yuanyuan SHI,Liping XIA,Lin SHI,Wei WU,Ling. LUO. The role of NLRP3 signaling pathway in allergic rhinoconjunctivitis[J]. The Journal of Practical Medicine, 2024, 40(14): 1922-1927.

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组别	挠鼻次数	眼部分泌物
对照组	0.500 ± 0.535	0.125 ± 0.354
实验组	2.600 ± 0.516^*	2.100 ± 0.738^*
NLRP3^-/-组	2.500 ± 0.527^*	1.700 ± 0.675^*
F值	43.595	23.802
P值	0.000	0.000

组别	鼻部过敏出现时间	眼部过敏出现时间
实验组	10.500 ± 1.080	20.300 ± 2.058
NLRP3^-/-组	13.400 ± 1.955^*	20.900 ± 2.132
t值	-4.106	-0.640
P值	0.001	0.530

组别	OVA-sIgE（ng/mL）	IL-4（pg/mL）	IL-17（pg/mL）	IL-18（pg/mL）
对照组	18.368 ± 1.667	56.552 ± 3.781	24.298 ± 3.640	57.798 ± 2.547
实验组	29.890 ± 2.012^*	98.447 ± 6.870^*	37.882 ± 4.324^*	87.183 ± 2.451^*
NLRP3^-/-组	23.462 ± 1.912^*a	95.383 ± 17.728^*	34.638 ± 3.924^*	75.032 ± 6.196^*△
F值	84.585	86.505	19.132	191.645
P值	0.000	0.000	0.000	0.000

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