Lin Lin, Dai Fei, Sun Tingyu
Department of Otorhinolaryngology Head and Neck Surgery, Huashan Hospital of Fudan University, Shanghai 200040, China.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2016 Jan;51(1):43-9. doi: 10.3760/cma.j.issn.1673-0860.2016.01.007.
The aim of the study was to investigate whether Orai1 antibody intraperitoneal injection could improve the condition of allergic rhinitis (AR) in mice.
Twenty-four BALB/C mice (SPF grade) were classified into 4 groups (AR group, Control group, Experimental group 1 and experimental group 2) according to a random number table. A mouse model of AR was established (Control group was established by phosphate buffered solution), and experimental group 1 and Experimental group 2 were established through intraperitoneal injection of 100 μg and 150 μg Orai1 antibody respectively. The number of sneezing and rubbing and eosinophilia in mice were assessed after different doses of Orai1 antibody intraperitoneal injection were applied. Then Orai1 protein and its mRNA in nasal mucosa, histomine, eosionphil cation protein (ECP), interlukin (IL)-1β, IL-4, IL-5 and IL-6 and their mRNA in nasal lavage fluid (NLF) and nasal mucosa were evaluated using enzyme linked immunosorbent assay (ELISA) and real-time reverse transcription-polymerase chain reaction (real-time RT-PCR). Furthermore, Orai1 protein and its mRNA in Th2 cells in peripheral blood, IL-4 and IL-5 in peripheral serum and their mRNAs in Th2 cells were also examined through ELISA and real-time RT-PCR. The data were analyzed by a statistical software of Graph Pad Prism 5.
There were significant differences in sneezing, nasal rubbing and local invading eosinophils in nasal mucosa after the treatment (t100 μg=7.88, t100 μg=9.92, t100 μg=4.30, respectively; t150 μg=16.43, t150 μg=16.31, t150 μg=9.35, respectively, all P-values<0.01). The Orai1 antibody intervention decreased contents of Orai1 in nasal mucosa, histomine, ECP, IL-1β, IL-4, IL-5 and IL-6. The contents of experimental group 1 were (0.186±0.015) μg/ml, (6.618±0.180) ng/ml, (2.555±0.031) ng/ml, (85.26±2.94) pg/ml, (55.12±1.21) pg/ml, (58.45±2.11) pg/ml and (77.12±2.13) pg/ml, respectively. The contents of experimental group 2 were (0.089±0.003) μg/ml, (4.501±0.310) ng/ml, (1.260±0.017) ng/ml, (48.49±2.12) pg/ml, (33.15±0.87) pg/ml, (38.24±0.95) pg/ml and (51.72±0.81) pg/ml, respectively. The differences were siginificant between group 1, group 2 and AR group(t value was 3.29, 10.44, 9.45, 17.53, 74.53, 87.06, 3.98; 8.54, 13.32, 23.00, 20.89, 80.73, 103.70, 13.34, all P<0.01). However, there were no significant differences in Orai1 protein and its mRNA in peripheral Th2 cells, IL-4 and IL-5 in peripheral serum and their mRNAs in Th2 cells (all P-values>0.05). In addition, the effect of 150 μg Orai1 antibody treatment was better than 100 μg one (all P-values<0.05).
Orai1 antibody intraperitoneal injection can improve the symptoms of AR mice, and alleviate the condition of allergic inflammation. Orai1 may become a novel aim in the AR study.
本研究旨在探讨腹腔注射Orai1抗体是否能改善小鼠过敏性鼻炎(AR)的病情。
将24只BALB/C小鼠(SPF级)按随机数字表法分为4组(AR组、对照组、实验组1和实验组2)。建立AR小鼠模型(对照组用磷酸盐缓冲溶液建立),实验组1和实验组2分别通过腹腔注射100μg和150μg Orai1抗体建立。在腹腔注射不同剂量的Orai1抗体后,评估小鼠的喷嚏次数、蹭鼻次数和嗜酸性粒细胞增多情况。然后采用酶联免疫吸附测定(ELISA)和实时逆转录-聚合酶链反应(实时RT-PCR)评估鼻黏膜中Orai1蛋白及其mRNA、组胺、嗜酸性粒细胞阳离子蛋白(ECP)、白细胞介素(IL)-1β、IL-4、IL-5和IL-6及其在鼻灌洗液(NLF)和鼻黏膜中的mRNA。此外,还通过ELISA和实时RT-PCR检测外周血Th2细胞中Orai1蛋白及其mRNA、外周血清中IL-4和IL-5及其在Th2细胞中的mRNA。数据用Graph Pad Prism 5统计软件分析。
治疗后,小鼠的喷嚏次数、鼻摩擦次数和鼻黏膜局部浸润嗜酸性粒细胞数有显著差异(t100μg分别为7.88、9.92、4.30;t150μg分别为16.43、16.31、9.35,所有P值<0.01)。Orai1抗体干预降低了鼻黏膜、组胺、ECP、IL-1β、IL-4、IL-5和IL-6中Orai1的含量。实验组1的含量分别为(0.186±0.015)μg/ml、(6.618±0.180)ng/ml、(2.555±0.031)ng/ml、(85.26±2.94)pg/ml、(55.12±1.21)pg/ml、(58.45±2.11)pg/ml和(77.12±2.13)pg/ml。实验组2的含量分别为(0.089±0.003)μg/ml、(4.501±0.310)ng/ml、(1.260±0.017)ng/ml、(48.49±2.12)pg/ml、(33.15±0.87)pg/ml、(38.24±0.95)pg/ml和(51.72±0.81)pg/ml。组1、组2与AR组之间差异有统计学意义(t值分别为3.29、10.44、9.45、17.53、74.53、87.06、3.98;8.54、13.32、23.00、20.89、80.73、103.70、13.34,均P<0.01)。然而,外周Th2细胞中Orai1蛋白及其mRNA、外周血清中IL-4和IL-5及其在Th2细胞中的mRNA无显著差异(所有P值>0.05)。此外,150μg Orai1抗体治疗效果优于100μg(所有P值<0.05)。
腹腔注射Orai1抗体可改善AR小鼠症状,减轻过敏性炎症病情。Orai1可能成为AR研究的新靶点。