椒目油A2对哮喘小鼠肺组织嗜酸性粒细胞凋亡的影响  

Effects of zanthoxylum seed oil A2 on eosinophil apoptosis in lung tissue of asthmatic mice

作　　者：王俊琴[1] 王胜春[1] 赵慧平[1] 刘明义[1] 

机构地区：[1]第四军医大学西京医院药剂科,西安710032

出　　处：《中国新药杂志》2010年第23期2174-2180,共7页Chinese Journal of New Drugs

基　　金：陕西省中医管理局科学基金项目(2005-076)

摘　　要：目的:观察椒目油A2(zanthoxylum seed oil A2,ZSOA2)对卵蛋白(ovalbumin,OVA)诱导哮喘小鼠不同时间点肺组织嗜酸性粒细胞(Eos)凋亡的影响。方法:BALB/c小鼠腹腔注射0.2 mL20%Al(OH)3和10%OVA的混合液致敏,然后经呼吸道滴入50μLOVA混合溶液(4 g OVA/0.01 mo.lL-1磷酸盐缓冲液)制备小鼠哮喘模型。滴入OVA混合溶液后24 h,48 h,3 d,7 d和14 d分别处死小鼠。伊红(HE)染色法检测肺组织病理形态学变化;末端脱氧核苷酸转移酶介导的带生物素dUTP切口末端标记技术(TUNEL)检测肺组织Eos的凋亡情况;原位杂交(ISH)法测定肺组织中白介素-5(IL-5)、嗜酸粒细胞趋化因子(eotaxin,EON)mRNA阳性粒细胞表达数;Western Blot法测定肺组织Fas,Fas L,Bax,BcL-2,Caspase-3,9,TNF-R1,p-JNK和c-jun信号通路蛋白的表达。结果:椒目油A2组能显著增加各时间点哮喘小鼠肺组织内Eos的凋亡率,降低IL-5,EON mRNA阳性粒细胞数,上调肺组织中各时间点Fas L和TNF-R1蛋白的表达,降低c-jun和p-JNK蛋白表达。结论:椒目油A2增加粒细胞凋亡率与降低IL-5,EON mRNA阳性粒细胞数、上调肺组织中Fas L和TNF-R1蛋白的表达有关。Objective： To observe the effects of zanthoxylum seed oil A2 （ZSOA2） on eosinophiI apoptosis at various time points after ovalbumin （OVA）-induced asthma in mice. Methods： BALB/c mice were sensitized by 0.2 mL mixture of 20% albumin hydroxide and 10% OVA intraperitoneally on day 1, then challenged by intra- tracheal drops of 50 μL OVA solution （4 g OVA in 0. 01 molvL^-1 phosphate buffer） to induce asthma. At 24 h, 48 h, 3 d, 7 d and 14 d after challenge, morphological changes in lung were assessed with HE staining. The percentage of eosinophil apoptosis in lung was detected by TUNEL; the mRNA expression of IL-5 and eotaxin（EON） in granulocytes were determined with in situ hybridization; the expression levels of Fas, Fas L, Bax, BcL-2, Caspase- 3 and -9, TNF-R1 , p-JNK, and c-jun in lung were determined by Western Blot. Results： ZSOA2 significantly increased eosinophil apoptosis and decreased the count of ceils that expressed mRNA of IL-5 and EON in lung at each time point. It also increased protein expressions of Fas L and TNF-R1, and decreased protein expression of c-jun and p-JNK in lung at each time point. Conclusion：ZSOA2 increases apoptosis of granulocytes in lung tissue of asthmatic mice, which is associated with up-regulated protein expressions of Fas L and TNF-R1.

关 键 词：椒目油A2 支气管哮喘 凋亡 嗜酸性粒细胞 

分 类 号：R284.1[医药卫生—中药学]