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1.北京中医药大学 深圳医院, 广东 深圳 518172
2.深圳市龙岗区耳鼻咽喉医院, 广东 深圳 518172
周才杰,博士,副主任中药师,从事中药防治过敏性疾病的临床和基础研究,E-mail:azhou828@163. com
修回日期:2019-10-21,
网络出版日期:2020-03-09,
纸质出版日期:2020-08-05
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周才杰,王小清,赵九洲等.玉屏风散对过敏性鼻炎小鼠GATA3,Foxp3表达的影响[J].中国实验方剂学杂志,2020,26(15):8-13.
ZHOU Cai-jie,WANG Xiao-qing,ZHAO Jiu-zhou,et al.Effect of Yupingfeng San on Expressions of GATA3,Foxp3 in Allergic Rhinitis Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(15):8-13.
周才杰,王小清,赵九洲等.玉屏风散对过敏性鼻炎小鼠GATA3,Foxp3表达的影响[J].中国实验方剂学杂志,2020,26(15):8-13. DOI: 10.13422/j.cnki.syfjx.20201104.
ZHOU Cai-jie,WANG Xiao-qing,ZHAO Jiu-zhou,et al.Effect of Yupingfeng San on Expressions of GATA3,Foxp3 in Allergic Rhinitis Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(15):8-13. DOI: 10.13422/j.cnki.syfjx.20201104.
目的
2
研究玉屏风散对卵清蛋白(OVA)诱导的过敏性鼻炎(AR)小鼠肺组织中GATA结合蛋白(GATA3)和叉头转录因子3(Foxp3)蛋白表达水平的影响,初步探讨玉屏风散治疗AR的作用机制。
方法
2
采用OVA+Al(OH)
3
腹腔注射致敏,然后OVA滴鼻激发的方法建立过敏性鼻炎小鼠模型,将小鼠分为正常组、模型组、氯雷他定组(3 mg·kg
-1
)和玉屏风散组(6.5 g·kg
-1
)。灌胃给药3周后,苏木素-伊红(HE)染色观察鼻黏膜组织中的肥大细胞、嗜酸性粒细胞、中性粒细胞等炎性细胞的浸润情况;酶联免疫吸附测定(ELISA)检测各组小鼠外周血中OVA特异性免疫球蛋白(Ig)E及白细胞介素-4(IL-4),IL-5,
γ
-干扰素(INF-
γ
)等细胞因子水平;蛋白免疫印迹法(Western blot)检测小鼠肺组织中GATA3和Foxp3蛋白的表达。
结果
2
AR模型小鼠出现挠鼻、喷嚏、流涕等症状;鼻黏膜切片HE染色可见AR小鼠鼻黏膜纤毛大量脱落,组织间质水肿明显,小血管扩张,大量嗜酸性细胞、淋巴细胞、浆细胞浸润。而玉屏风散灌胃给药可明显改善过敏性鼻炎小鼠的鼻部症状;鼻黏膜上皮结构完整,排列均匀,未见明显组织间隙水肿和血管扩张,炎性细胞浸润情况明显减轻。与正常组比较,AR模型组小鼠外周血中OVA特异性IgE以及IL-4,IL-5等细胞因子水平显著升高(
P
<
0.01),而INF-
γ
水平显著降低(
P
<
0.01);与模型组比较,氯雷他定和玉屏风散灌胃给药能显著降低AR小鼠外周血中OVA特异性IgE以及IL-4,IL-5等细胞因子水平(
P
<
0.05),同时上调INF-
γ
水平(
P
<
0.05,
P
<
0.01);Western blot结果显示,与正常组比较,AR模型组小鼠肺组织中GATA3蛋白的表达显著增加,而Foxp3蛋白的表达明显降低(
P
<
0.05,
P
<
0.01);与模型组比较,玉屏风散能显著降低GATA3蛋白的表达,并显著升高Foxp3蛋白的表达(
P
<
0.01)。
结论
2
玉屏风散对OVA诱导的过敏性鼻炎具有改善作用;玉屏风散可能通过调节Th2/Treg平衡等机制调节机体的免疫反应。
Objective
2
To study the effect of Yupingfeng San (YPFS) on the expressions of GATA binding protein 3 (GATA3) and forkhead transcription factor3 (Foxp3) in ovalbumin (OVA)-induced lung tissue of allergic rhinitis (AR) mice, and explore the mechanism of YPFS on AR.
Method
2
The allergic rhinitismice model was established by intraperitoneally injecting with OVA and Al (OH)
3
, and challenged with OVA intranasally. The mice were divided into four groups: normal,model,chloretadine(3 mg·kg
-1
) and YPFS(6.5 g·kg
-1
) group, the corresponding drugs were orally administrated for three weeks. At the end of administration,the infiltration of inflammatory cells, such as mast cells, eosinophils and neutrophils in nasal mucosa, were observed by htoxylin eosin (HE) staining. The serum concentrations of OVA-specific IgE and cytokines [interleukin-4(IL-4), IL-5 and
γ
-interferon (INF-
γ
)] were determined by enzyme-linked immune sorbent assay (ELISA). The expressions of GATA3 and Foxp3 proteins in nasal mucosa tissue were detected by Western blot.
Result
2
The AR mice had such symptoms as scratching, sneezing and running nose. Nasal mucosa section by HE staining showed significant desquamation of AR mouse nasal mucosa cilia, obvious tissue stromal edema, telangiectasia, and a large number of eosinophilic cells, lymphocytes, plasma cells infiltration. YPFS obviously improved nasal symptoms of allergic rhinitis mice. Nasal mucosa epithelial structure was complete and arranged evenly, with no obvious tissue clearance edema and vasodilation, and inflammatory cell infiltration was significantly reduced. Compared with normal group, the levels of OVA specific IgE, IL-4 and IL-5 in peripheral blood of AR model group were significantly higher(
P
<
0.01), and the INF-
γ
level was significantly lower (
P
<
0.01). Compared with AR model group, the administration of chloretadine and YPFS can significantly reduce the level of OVA specific IgE and IL-4, IL-5, and increase the level of INF-
γ
in AR mice peripheral blood (
P
<
0.05,
P
<
0.01). Western blot results showed that compared with normal group, GATA3 protein expression was significantly increased, while Foxp3 protein expression was significantly decreased in AR model group (
P
<
0.05,
P
<
0.01). Compared with AR model group, YPFS inhibited GATA3, and promoted Foxp3 protein expression (
P
<
0.01).
Conclusion
2
YPFS has an effect in alleviating OVA-induced allergic rhinitis.YPFS may modulate the immune response by regulating the balance of Th2/Treg cells.
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