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肠球菌益生菌制剂对小鼠实验性变应性鼻炎的抗过敏作用

[Anti-allergic effects of the probiotic preparations of enterococcus on experimental allergic rhinitis in mice].

作者信息

Zhu Lu-Ping, Zhang Qing-Zhao, Shimada Takashi, Enomoto Tadao, Cheng Lei

机构信息

Department of Otorhinolaryngology, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210011, China.

出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2013 Jul;48(7):555-62.

PMID:24313204
Abstract

OBJECTIVE

The aim of this study was to investigate the anti-allergic effects of lysozyme/heat-treated Enterococcus faecalis FK-23 (LFK) and heat-treated Enterococcus faecium sp. TN-3 (TN) on experimental allergic rhinitis (AR).

METHODS

A total of twenty-four BALB/c mice were divided into four groups randomly: (1) positive control group, (2) LFK-fed group, (3) TN-fed group, and (4) negative control group. To establish the murine AR model, intraperitoneal injection and nasal drip with ovalbumin (OVA) were performed. Saline was used instead of OVA for negative control. Probiotic preparations of LFK and TN were orally administrated for 42 days [60 mg (0.5 ml)/d] in LFK-fed and TN-fed mice, respectively. The positive and negative control mice received saline orally for 42 days. Nasal rubbing and sneezing were monitored on d 21, d 27, and d 35. After the final challenge, mice were sacrificed on d 42, and eosinophilic infiltration into the nasal mucosa was quantified (H&E stain). IFN-gamma, IL4 and OVA-specific IgE levels in the sera and splenocyte culture supernatants were determined by ELISA kits.

RESULTS

Nasal rubbing of LFK-fed mice was significantly reduced compared to the positive control group on day 27 (t = 2.95, P = 0.028). Both in the LFK-fed and TN-fed mice, nasal rubbing (t value was 3.75 and 3.06, P value was 0.005 and 0.011, respectively) and sneezing (t value was 2.56 and 3.35, P value was 0.038 and 0. 01, respectively) were significantly decreased compared to the positive control group on d 35. The H&E strain section of nasal tissue showed that eosinophil influx into the nasal mucosa decreased significantly both in the LFK-fed and TN-fed mice compared to the positive control group on day 42 (t value was 3.44 and 2.97, P value was 0.014 and 0.025, respectively); however, the LFK-fed mice and TN-fed mice had significant eosinophil influx into the nasal mucosa than that in the negative control group (t value was 2.54 and 3.39, P value was 0.044 and 0.015, respectively). There were no significant differences in the serum levels of IL-4 and OVA-specific IgE, as well as the levels of IFN-gamma and IL-4 in splenocyte culture supernatants between the LFK-fed group and positive control group on d 42 (all P > 0.05). Interestingly, the TN-fed mice had significantly higher serum levels of IFN-gamma compared to the LFK-fed mice [TN-fed mice: (27.07 +/- 3.83) pg/ml, LFK-fed mice: (14.83 +/- 0.99) pg/ml; Z = 2.49, P = 0.016], but not the negative control group [negative control group: (37.12 +/- 1.65) pg/ml; Z = 1.18, P = 0.343] on day 42. The serum levels of IL-4 were significantly lower in the TN-fed mice than those in the positive control group [TN-fed mice: (34.48 +/- 7.53) pg/ml, positive control group: (58.68 +/- 6.59) pg/ml; Z = 2.11, P = 0.035]; however, the levels were significantly higher in the TN-fed mice than those in the negative control group [negative control group: (20.22 +/- 1.75) pg/ml; Z = 2. 31, P = 0.021]. The TN-fed mice had significantly higher levels of IFN-gamma in splenocyte culture supernatants compared to the LFK-fed mice (Z = 2.72, P = 0.03) and the positive control group (Z = 2.30, P = 0.029), whilst the splenocyte culture supernatant levels of IL-4 (Z = 2.12, P = 0.034) and the serum levels of OVA-specific IgE (Z = 2.31, P = 0.021) were significantly lower in the TN-fed mice compared to the positive control mice.

CONCLUSIONS

It is suggested that oral administration of probiotic LFK or TN may alleviate nasal symptoms and reduce nasal eosinophilia in the murine AR model. TN supplementation has obviously regulatory effects on the cytokine levels of IFN-gamma and IL-4, and significantly inhibitory effects on antigen-specific IgE levels.

摘要

目的

本研究旨在探讨溶菌酶/热处理粪肠球菌FK-23(LFK)和热处理屎肠球菌sp.TN-3(TN)对实验性变应性鼻炎(AR)的抗过敏作用。

方法

将24只BALB/c小鼠随机分为四组:(1)阳性对照组,(2)LFK喂养组,(3)TN喂养组,(4)阴性对照组。通过腹腔注射和滴鼻卵清蛋白(OVA)建立小鼠AR模型。阴性对照用生理盐水代替OVA。分别对LFK喂养组和TN喂养组的小鼠口服LFK和TN的益生菌制剂42天[60mg(0.5ml)/天]。阳性和阴性对照小鼠口服生理盐水42天。在第21、27和35天监测鼻擦和打喷嚏情况。末次激发后,于第42天处死小鼠,对鼻黏膜嗜酸性粒细胞浸润进行定量(苏木精-伊红染色)。采用ELISA试剂盒测定血清和脾细胞培养上清液中IFN-γ、IL-4和OVA特异性IgE水平。

结果

与阳性对照组相比,LFK喂养组小鼠在第27天鼻擦次数显著减少(t = 2.95,P = 0.028)。在第35天,LFK喂养组和TN喂养组小鼠的鼻擦次数(t值分别为3.而LFK喂养组和TN喂养组小鼠鼻黏膜嗜酸性粒细胞浸润均显著高于阴性对照组(t值分别为2.54和3.39,P值分别为0.044和0.015)。在第42天,LFK喂养组与阳性对照组血清IL-4水平、OVA特异性IgE水平以及脾细胞培养上清液中IFN-γ和IL-4水平均无显著差异(所有P>0.05)。有趣的是,在第42天,TN喂养组小鼠血清IFN-γ水平显著高于LFK喂养组[TN喂养组:(27.07±3.83)pg/ml,LFK喂养组:(14.83±0.99)pg/ml;Z = 2.49,P = 0.016],但与阴性对照组相比无显著差异[阴性对照组:(37.12±1.65)pg/ml;Z = 1.18,P = 0.343]。TN喂养组小鼠血清IL-4水平显著低于阳性对照组[TN喂养组:(34.48±7.53)pg/ml,阳性对照组:(58.68±6.59)pg/ml;Z = 2.11,P = 0.035];然而,TN喂养组小鼠血清IL-4水平显著高于阴性对照组[阴性对照组:(20.22±1.75)pg/ml;Z = 2.31,P = 0.021]。与LFK喂养组小鼠(Z = 2.72,P = 0.03)和阳性对照组(Z = 2.30,P = 0.029)相比,TN喂养组小鼠脾细胞培养上清液中IFN-γ水平显著升高,而TN喂养组小鼠脾细胞培养上清液中IL-4水平(Z = 2.12,P = 0.034)和血清OVA特异性IgE水平(Z = 2.31,P = 0.021)显著低于阳性对照小鼠。

结论

提示口服益生菌LFK或TN可减轻小鼠AR模型的鼻部症状并减少鼻黏膜嗜酸性粒细胞浸润。补充TN对IFN-γ和IL-4细胞因子水平有明显调节作用,对抗原特异性IgE水平有显著抑制作用。

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