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1.北京中医药大学 第三附属医院,北京 100029
2.北京中医药大学,北京 100029
候丹,博士,主治医师,从事中医药防治肺系疾病的研究,E-mail:859669724@qq.com
崔红生,博士,教授,主任医师,博士生导师,从事中医药防治肺系疾病的研究,E-mail:hshcui@sina.com
收稿日期:2021-06-27,
网络出版日期:2021-10-12,
纸质出版日期:2022-01-20
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候丹,黄帅阳,吕明圣等.桑梅止咳颗粒对咳嗽变异性哮喘大鼠气道炎症反应的作用及机制[J].中国实验方剂学杂志,2022,28(02):62-66.
HOU Dan,HUANG Shuai-yang,LYU Ming-sheng,et al.Effect and Mechanism of Sangmei Zhike Granule on Airway Inflammation in Rats with Cough Variant Asthma[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):62-66.
候丹,黄帅阳,吕明圣等.桑梅止咳颗粒对咳嗽变异性哮喘大鼠气道炎症反应的作用及机制[J].中国实验方剂学杂志,2022,28(02):62-66. DOI: 10.13422/j.cnki.syfjx.20212223.
HOU Dan,HUANG Shuai-yang,LYU Ming-sheng,et al.Effect and Mechanism of Sangmei Zhike Granule on Airway Inflammation in Rats with Cough Variant Asthma[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):62-66. DOI: 10.13422/j.cnki.syfjx.20212223.
目的
2
观察桑梅止咳颗粒对咳嗽变异性哮喘(CVA)大鼠炎症反应的调节作用及机制。
方法
2
6周龄雄性SD大鼠随机分为正常组、模型组、中药组(桑梅止咳颗粒2.48 g·kg
-1
灌胃);模型组、中药组分别在第1,8天腹腔注射10%卵蛋白(OVA),氢氧化铝混合液+第15天开始1%OVA溶液雾化激发建立CVA模型;第15天各组连续干预2周。实验结束,检测大鼠肺功能,外周血炎细胞计数,血清白细胞介素(IL)-4,IL-5,IL-10含量,肺组织苏木素-伊红(HE)染色;蛋白免疫印迹法(Western blot)检测肺组织核转录因子-
κ
B(NF-
κ
B)抑制蛋白
α
(I
κ
B
α
),NF-
κ
B蛋白表达。
结果
2
与正常组比较,模型组大鼠0.1 s用力呼气容积(FEV
0.1
),0.1 s内平均流速(FEV
0.1
/FVC),呼出50%FVC量时流速(FEF50%)明显降低(
P
<
0.05,
P
<
0.01);外周血白细胞计数、嗜酸性粒细胞计数显著升高(
P
<
0.01);血清IL-4,IL-5,IL-10含量显著升高(
P
<
0.01)。HE染色显示模型组支气管可见上皮细胞脱落,气道狭窄,黏液腺增生扩张,各层结构紊乱,细胞排列混乱,大量炎细胞浸润。与正常组比较,模型组肺组织NF-
κ
B p65蛋白表达显著升高(
P
<
0.01),I
κ
B
α
蛋白表达显著降低(
P
<
0.01)。与模型组比较,中药组大鼠FEV
0.1
,FEV
0.1
/FVC,FEF50%明显升高(
P
<
0.05);外周血白细胞计数、嗜酸性粒细胞计数显著降低(
P
<
0.01);血清IL-4,IL-5,IL-10含量显著降低(
P
<
0.01);HE染色显示中药组支气管黏膜损伤较轻,炎细胞浸润、腺体增生、上皮变性坏死等明显减轻。与模型组比较,中药组肺组织NF-
κ
B p65蛋白表达明显降低(
P
<
0.05),I
κ
B
α
蛋白表达明显升高(
P
<
0.05)。
结论
2
桑梅止咳颗粒具有改善CVA大鼠肺功能、抑制气道炎症反应的作用,机制可能与调控大鼠肺I
κ
Bα/NF-
κ
B表达相关。
Objective
2
To explore the effect and mechanism of Sangmei Zhike granule (SMZK) on airway inflammation in rats with cough variant asthma(CVA).
Method
2
Six-week-old male SD rats were randomly divided into normal group, model group, and SMZK (2.48 g·kg
-1
) group. The rats in the model group and the SMZK group received intraperitoneal injection of a mixed solution containing 10% ovalbumin (OVA) and aluminium hydroxide on the 1
st
and 8
th
days and aerosol inhalation of 1% OVA solution from the 15
th
day for CVA model induction. The intervention lasted for two weeks from the 15
th
day. At the end of animal manipulation, the lung function was detected and inflammatory cells in the peripheral blood were counted. The serum interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-10 (IL-10) levels were determined. Hematoxylin-eosin (HE) staining was performed on the lungs. Western blot was used to detect the protein expression of nuclear factor kappa-B (NF-
κ
B) and its inhibitor
α
(I
κ
B
α
) in lung tissues.
Result
2
Compared with the normal group, the model group showed reduced forced expiratory volume in the first 0.1 second (FEV
0.1
),FEV
0.1
/forced vital capacity (FVC),and forced expiratory flow 50% (FEF50%) (
P
<
0.05,
P
<
0.01), increased white blood cells and eosinophils (
P
<
0.01), and up-regulated serum IL-4, IL-5, and IL-10 (
P
<
0.01). As revealed by HE staining, the model group displayed shed epithelial cells of the bronchus, airway stenosis, hyperplasia and expansion of mucous glands, disarrangement of layer structures, disorderly arranged cells, and extensive infiltration of inflammatory cells. The protein expression of NF-
κ
B p65 was higher (
P
<
0.01) and that of I
κ
B
α
was lower (
P
<
0.01) in the lung tissues of the model group than that in the normal group. Compared with the model group, the SMZK group showed increased FEV
0.1
,FEV
0.1
/FVC,and FEF50% (
P
<
0.05), decreased white blood cells and eosinophils in the peripheral blood (
P
<
0.01), and declining serum IL-4, IL-5, IL-10 (
P
<
0.01). HE staining demonstrated mild bronchial mucosal injury and relieved inflammatory cell infiltration, gland hyperplasia, and epithelial degeneration and necrosis in the SMZK group. The protein expression of NF-
κ
B p65 was decreased (
P
<
0.05) and that of I
κ
B
α
was increased (
P
<
0.05) in lung tissues of the SMZK group than that in the model group.
Conclusion
2
SMZK can improve lung function and inhibit airway inflammation in rats with CVA. The underlying mechanism may be related to the regulation of I
κ
B
α
/NF-
κ
B protein expression in the lungs.
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