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[幼年大鼠哮喘模型中嗜酸性粒细胞凋亡、fas mRNA和bcl-2 mRNA表达及牛膝多糖的影响]

[Eosinophils apoptosis, fas mRNA and bcl-2 mRNA expressions in asthma model of young rat and effects of achyranthes bidentata polysaccharides].

作者信息

Li Chang-chong, Hu Xiao-guang, Zhang Wei-xi, Xie Li-wei, Zhang Hai-yan, Dong Lin, Cai Xiao-hong, Wu Rong-xi, Zhang Zheng-xia, He Qiu-sha

机构信息

Division of Pulmonology, Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical College, Wenzhou 325027, China.

出版信息

Zhonghua Er Ke Za Zhi. 2003 Sep;41(9):657-60.

PMID:14733803
Abstract

OBJECTIVE

Asthma is a chronic respiratory tract disorder characterized by airway hyperreaction (AHR), persistent airway inflammation, high serum IgE, overproduction of IL-4, IL-5 and IL-13 by allergen-specific Th2 cells. The morbidity and mortality of asthma have continued to increase despite the use of currently available therapeutic agents. The reputed effects of traditional Chinese medicines (TCMs) have led to increasing use of TCMs for treatment of asthma throughout the world. The aims of this study were to investigate in asthma model of young rat the mRNA expressions of apoptotic gene fas and bcl-2, eosinophils (EOS) apoptosis in airway, and effects of achyranthes bidentata polysaccharides (ABPS), a group of polysaccharides extracted from TCM Achyranthes bidentata blume, on treatment of asthma.

METHODS

Fifty Sprague-Dawley (SD) rats were divided into five groups, 10 rats per group. Asthma in rats was induced by intraperitioneal sensitization and challenge with nebulized ovalbumin (OVA). A pretreatment with ABPS [50 mg/(kg x d)] was done according to three different schedules: consecutively 3 days at sensitization (T1), at challenge (T2) or both of the two periods (T3). Sham-treated rats (A) and naive rats (C) served as controls. The animals were sacrificed 24 hours after the last challenge. The mRNA expression of bcl-2 and fas in eosinophils presenting in airway and the apoptosis of eosinophils in airway were assessed by using in situ hybridization with oligonucleotide probe and TUNEL methods, respectively.

RESULTS

(1) Twenty-four hours after the last antigen challenge, the mRNA expression of fas in eosinophils presenting in airway significantly decreased in group A [(43.4 +/- 10.0)%] compared with that in group C [(73.2 +/- 11.9)%] (P < 0.01). ABPS could increase the fas mRNA expression significantly in all the three groups [(59.0 +/- 8.1)%, (57.5 +/- 9.6)%, (76.2 +/- 2.7)%], compared with that in group A (P < 0.05, P < 0.05, P < 0.01, respectively). The expression of the bcl-2 mRNA in group C was (47.9 +/- 8.7)%, it was elevated to (67.4 +/- 7.3)% in group A (P < 0.01). The expression of the bcl-2 mRNA in ABPS treated T1 and T3 groups was significantly lowered [(57.7 +/- 12.7)%, (57.3 +/- 6.8)%, P < 0.05], but not in T2 group [(72.4 +/- 6.7)%]. (2) In group A, the EOS presenting in the airway increased significantly, but there were few apoptotic EOS; the percentage of apoptotic eosinophil was distinctly lower in group A than that in group C [(5.3 +/- 2.2)% vs. (15.9 +/- 2.4)%, P < 0.01]. Compared with that in group A, the eosinophil apoptosis ratio in those ABPS treated groups T1, T3 was evidently elevated [(8.7 +/- 2.9)%, (9.8 +/- 2.2)%, P < 0.05, P < 0.05], but ABPS treated at challenge (T2) could not change the eosinophil apoptosis ratio significantly (P > 0.05).

CONCLUSION

(1) In asthmatic rat, the expressions of the genes fas and bcl-2 mRNA in EOS were changed evidently and the ratio of EOS apoptotosis reduced greatly. (2) ABPS could enhance the apoptosis of EOS by upregulating the expression of the genes fas and bcl-2 mRNA.

摘要

目的

哮喘是一种慢性呼吸道疾病,其特征为气道高反应性(AHR)、持续性气道炎症、血清IgE升高、变应原特异性Th2细胞过度产生白细胞介素-4(IL-4)、白细胞介素-5(IL-5)和白细胞介素-13。尽管使用了现有的治疗药物,哮喘的发病率和死亡率仍持续上升。中药(TCM)的显著疗效促使其在全球范围内用于哮喘治疗的应用不断增加。本研究旨在探讨在幼龄大鼠哮喘模型中凋亡基因fas和bcl-2的mRNA表达、气道嗜酸性粒细胞(EOS)凋亡情况,以及从中药牛膝中提取的一组多糖——牛膝多糖(ABPS)对哮喘治疗的作用。

方法

将50只Sprague-Dawley(SD)大鼠分为五组,每组10只。通过腹腔致敏和雾化卵清蛋白(OVA)激发诱导大鼠哮喘。根据三种不同方案用ABPS[50mg/(kg·d)]进行预处理:致敏期连续3天(T1)、激发期(T2)或两个时期均进行(T3)。假处理大鼠(A)和未处理大鼠(C)作为对照。在最后一次激发后24小时处死动物。分别采用寡核苷酸探针原位杂交和TUNEL法评估气道中嗜酸性粒细胞bcl-2和fas的mRNA表达以及气道嗜酸性粒细胞凋亡情况。

结果

(1)末次抗原激发后24小时,与C组[(73.2±11.9)%]相比,A组气道中嗜酸性粒细胞fas的mRNA表达显著降低[(43.4±10.0)%](P<0.01)。与A组相比,ABPS可使所有三组的fas mRNA表达显著增加[(59.0±8.1)%、(57.5±9.6)%、(76.2±2.7)%](分别为P<0.05、P<0.05、P<0.01)。C组bcl-2 mRNA表达为(47.9±8.7)%,A组升高至(67.4±7.3)%(P<0.01)。ABPS处理的T1组和T3组bcl-2 mRNA表达显著降低[(57.7±12.7)%、(57.3±6.8)%,P<0.05],但T2组未降低[(72.4±6.7)%]。(2)A组气道中EOS显著增加,但凋亡的EOS很少;A组凋亡嗜酸性粒细胞百分比明显低于C组[(5.3±2.2)%对(15.9±2.4)%,P<0.01]。与A组相比,ABPS处理的T1组、T3组嗜酸性粒细胞凋亡率明显升高[(8.7±2.9)%、(9.8±2.2)%,P<0.05、P<0.05],但激发期用ABPS处理(T2)不能显著改变嗜酸性粒细胞凋亡率(P>0.05)。

结论

(1)在哮喘大鼠中,EOS中fas和bcl-2基因的mRNA表达明显改变,EOS凋亡率大幅降低。(2)ABPS可通过上调fas和bcl-2基因的mRNA表达增强EOS凋亡。

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