槐杞黄对OVA诱导哮喘模型大鼠嗜酸性细胞凋亡及免疫调节因子的影响

目的探讨槐杞黄对哮喘模型大鼠嗜酸性细胞凋亡、Th1/Th2/Th17免疫调节及相关炎症因子的影响。方法将60只SD大鼠随机分成正常对照组、模型组、地塞米松组、槐杞黄组、槐杞黄联合地塞米松组,每组12只。卵清蛋白(OVA)致敏激发复制大鼠哮喘模型,对照组和模型组大鼠给予生理盐水,槐杞黄组大鼠按0.4g·100g-1·d-1给予槐杞黄颗粒混悬液灌胃,地塞米松组大鼠按0.1g·100g-1·d-1给予地塞米松混悬液,槐杞黄联合地塞米松组大鼠给予槐杞黄(0.4g·100g-1·d-1)+地塞米松(0.1g·100g-1·d-1),连续给药15天后,测定各组大鼠血清免疫球蛋白E(IgE)、白介素5(IL-5)、白介素3(IL-3)、粒细胞巨噬细胞集落刺激因子(GM-CSF)及白介素17(IL-17)水平;肺泡盥洗液进行白细胞计数及分类;病理切片观察肺组织病理变化;实时定量-PCR测定肺组织Bcl-2、Bax及NF-κB mRNA表达。结果与模型组比较,地塞米松、槐杞黄以及槐杞黄联合地塞米松可显著降低哮喘大鼠血清IgE水平(57.14±5.66)ng/mL、(57.87±5.31)ng/mL、(46.68±6.67)ng/mL比(84.17±5.41)ng/mL,P<0.05,P<0.01];槐杞黄联合地塞米松可明显降低哮喘大鼠血清IL-5(23.56±6.60)pg/mL比(32.20±10.64)pg/mL,P<0.05]、IL-3 (2717.38±128.88)pg/mL比(4235.66±165.50)pg/mL,P<0.01]、GM-CSF(6.84±5.54)pg/mL比(13.20±2.01)pg/mL,P<0.01]及IL-17(252.25±15.14)pg/mL比(415.80±32.67)pg/mL,P<0.01]水平;地塞米松和槐杞黄联合地塞米松可降低哮喘模型大鼠肺泡盥洗液白细胞总数(83.41±4.65)、(63.7±6.10)比(169.40±11.18),P均<0.05],以及嗜酸粒细胞(EOS)(1.05±0.02)、(0.71±0.04)比(7.01±0.18),P均<0.05]和淋巴细胞(LYM)(7.91±0.09)、(8.18±0.23)比(17.91±0.11),P均<0.05]百分比。给药组均可缓解大鼠肺组织肺泡壁及气道周围炎症细胞浸润的现象。地塞米松、槐杞黄以及槐杞黄联合地塞米松均可明显降低哮喘模型大鼠肺组织Bcl-2/Bax mRNA相对表达量(0.92±0.24)、(1.65±0.28)、(0.78±0.44)比(6.08±2.31),P均<0.01]。槐杞黄以及槐杞黄联合地塞米松可降低哮喘大鼠肺组织NF-κB mRNA相对表达量(1.19±0.11)、(1.10±0.15)比(1.46±0.08),P均<0.05]。结论槐杞黄辅助治疗可减轻哮喘模型大鼠气道炎症,其机制可能与纠正Th1/Th2/Th17免疫失衡,促进嗜酸性细胞凋亡,减少哮喘发作炎症因子水平相关。

Effect of Huaiqihuang on OVA-induced apoptosis of eosinophil cells and immunoregulatory factors in asthmatic rats

Objective To investigate the effect of Huaiqihuang on the apoptosis of eosinophil, Th1/Th2/Th17 immunomodulation and related inflammatory factors in asthmatic rats. Methods Sixty SD rats were randomly divided into normal control group, model group, dexamethasone group, Huaiqihuang group, Huaiqihuang combined with dexamethasone group, 12 rats in each group. Ovalbumin(OVA) sensitized and stimulated the construction of rat asthma model. The control group and model group rats were given normal saline. Rats in the Huaiqihuang group were given Huaiqihuang granule suspension by 0.4 g/100 g/d. Rats in the dexamethasone group were given dexamethasone suspension by 0.1 g/100 g/d. Rats in the Huaiqihuang combined with dexamethasone group were given 0.4 g/100 g/d Huaiqihuang and 0.1 g/100 g/d dexamethasone. Levels of serum immunoglobulin E(IgE), interleukin 5(IL-5), interleukin 3(IL-3), granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 17(IL-17) were measured after 15 days of continuous administration. Alveolar washing fluid was collected for white blood cell count and classification. Pathological section was made to observe the pathological changes of lung tissue. Real-time quantitative-PCR was used to determine mRNA expression of Bcl-2, Bax and NF-κB in lung tissues. Results Compared to the model group, Serum IgE level in asthmatic rats of dexamethasone group, Huaiqihuang group, and Huaiqihuang combined with dexamethasone group decreased significantly (57.14±5.66),(57.87±5.31),(46.68±6.67) vs(84.17±5.41)ng/mL, P<0.05, P<0.01]. So did in the Huaiqihuang combined with dexamethasone group for IL-5(23.56±6.60) vs(22.96±1.31)pg/mL, P<0.05], IL-3(2717.38±128.88) vs(2033.76±114.33)pg/mL, P<0.01], GM-CSF(6.84±5.54) vs(6.20±0.65)pg/mL, P<0.01], and IL-17(252.25±15.14) vs(289.64±21.64)pg/mL, P<0.05]. Also, in the Huaiqihuang group, and Huaiqihuang combined with dexamethasone group, total white blood cells in the alveolar washing fluid reduced(83.41±4.65) and(63.7±6.10) vs(169.40 ±11.18), P<0.05, respectively], eosinophils(EOS)count lessened(1.05±0.02) and(0.71±0.04) vs(7.01±0.18), P<0.05, respectively], and lymphocyte(LYM) dropped(7.91±0.09) and(8.18±0.23) vs(17.91±0.11), P<0.05, respectively]. The infiltration of inflammatory cells around the pulmonary alveolar wall and airway was relieved by the administration of different drug administration. The relative mRNA expression of Bcl-2/Bax in the lung tissues of dexamethasone group, Huaiqihuang group and the combination group rats reduced significantly(0.92±0.24),(1.65±0.28),(0.78±0.44) vs(6.08±2.31), P<0.01, respectively]. In the Huaiqihuang group, and Huaiqihuang combined with dexamethasone group, NF-κB mRNA expression decreased(1.19±0.11) and(1.10±0.15) vs(1.46±0.08), P<0.05]. Conclusion Adjuvant treatment of Huaiqihuang can alleviate airway inflammation in asthmatic rats, and its mechanism may be related to correcting Th1/Th2/Th17 immune imbalance, promoting eosinophil apoptosis, and reducing the level of inflammatory factors in asthma attacks.