氯雷他定对哮喘豚鼠白细胞介素5及嗜酸性粒细胞凋亡的影响

Effect of Loratadine on Expression of Interleukin-5 and Eosinophil Apoptosis in Asthmatic Guinea Pigs

目的研究氯雷他定对哮喘豚鼠支气管肺泡灌洗液(BALF)和血清IL-5的表达及肺组织嗜酸性粒细胞(EOS)凋亡延迟状态的影响。方法豚鼠32只随机分为正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组。经3次20 g·L-1卵清蛋白(OVA)腹腔注射致敏后,10 g·L-1OVA雾化吸入40 min诱发哮喘发作。氯雷他定低、高剂量组豚鼠分别予2 mg·kg-1·d-1、10mg·kg-1·d-1氯雷他定灌服14 d,每天1次;哮喘组、正常对照组给予9 g·L-1盐水作对照。ELISA法检测其血清和BALF中IL-5水平,罗氏TUNEL原位细胞凋亡检测试剂盒检测其凋亡细胞,HE染色切片随机计数每张切片下200个EOS中凋亡细胞数。结果1.正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组血清IL-5水平分别为(0.27±0.04)ng·L-1、(0.41±0.03)ng·L-1、(0.38±0.02)ng·L-1、(0.31±0.03)ng·L-1;BALF中IL-5水平分别为(0.10±0.01)ng·L-1、(0.38±0.04)ng·L-1、(0.33±0.05)ng·L-1、(0.23±0.09)ng·L-1,血清及BALF中IL-5水平4组间两两比较差异均有统计学意义(Pa<0.05)。2.正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组豚鼠肺组织EOS发生凋亡的细胞数分别为33个、95个、129个、182个,两两比较差异有统计学意义(χ2=114.5,P<0.05)。其中哮喘组与氯雷他定低剂量组间差异无统计学意义(χ2=10.0,P>0.008 3)。结论氯雷他定对IL-5相关的EOS凋亡延迟状态有抑制作用,能改善肺组织变应性炎症状态,对哮喘的控制具有潜在的积极作用。

Objective To study the influence of loratadine on the expression of interleukin - 5 （ IL - 5 ） in bronchoalveolar lavage fluid （BALF） and peripheral blood, and the eosinophil （EOS） apoptosis in asthmatic guinea pigs. Methods Thirty -two guinea pigs were ran- domly divided into the normal control group, the asthmatic group, the low - dose loratadine group and the high - dose loratadine group. The guinea pigs were peritoneal injected with 20 g·L^-1 ovalbumin（OVA） 3 times for allergization,inhaled 10 g·L^-1 OVA inhalation about 40 min to induce asthma attacks. Guinea pigs in low - dose loratadine group and high - dose loratadine group were drenched with 2 mg·L^-1 ·d^-1, 10 mg ·L^-1·d^-1 of loratadine about 14 days, once a day. The normal control group, the asthmatic group inhaled physiological saline. Enzyme linked immunosorbent assay was used to measure the contents of IL - 5 in BALF and serum, meanwhile, eosinophil apoptosis was also detected by terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling（TUNEL）. Combined with the hematoxylin -eosin stai- ning slices, the apoptosis cells in every 200 EOS in one slice were counted randomly. Results 1. The serum IL - 5 in the normal control group, asthmatic group, low - dose loratadine group and high - dose loratadine group were （0.27 ± 0.04 ） ng ·L^-1 （0.41 ~ 0.03 ） ng ·L^-1, （0.38~0.02） ng·L^-1,（0.31±0.03） ng·L^-1;IL-5 in BALF were respectively （ 0.10 ± 0. 01） ng ·L^-1,（0.38±0.04） ng· L-1, （ 0. 33±0.05 ） ng·L^-1, （0.23 ＋ 0.09 ） ng ·L^-1, comparison between every 2 groups, the differences were significant （ Pa 〈 0.05 ）. 2. The apoptosis cells in the 4 groups were 33,95,129,182, respectively,the differences between them were statistically significant（X^2 = 114.5,P 〈 0. 05 ） , but the difference between the asthmatic group and the low -dose loratadine group was not significant （X^2 = 10.0, P 〉 0.008 3 ）. Con- clusions Loratadine can reverse the IL -5 afforded delayed apoptosis, ameliorate the inflamatory state in lung tissue, and is of active thera- peutic value potentially in asthma management.