摘要
目的:探讨补肺汤对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)稳定期大鼠气道重塑的影响,求证补肺汤是否能通过TLR2/NF-κB信号转导通路来调节COPD大鼠免疫功能及抑制其气道炎症。方法:将84只大鼠随机分为假手术组、模型组、补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组、补肺汤免煎颗粒剂低剂量组、补肺汤饮片汤剂组和西药组,每组各12只。除假手术组以外其余各组均予以气道注射脂多糖联合烟熏的方法制作COPD动物模型,假手术组气道注射等量生理盐水,不予被动吸烟。造模成功后,第2天开始灌胃给药,假手术组、模型组给予10 m L·(kg·d)^(-1)生理盐水,西药组给予1.0 mg·(kg·d)^(-1)孟鲁司特钠,补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组、补肺汤免煎颗粒剂低剂量组分别给予[27.8 g·(kg·d)^(-1)、13.9 g·(kg·d)^(-1)、7.0 g·(kg·d)^(-1)]补肺汤免煎剂,补肺汤饮片汤剂组给予13.9 g·(kg·d)^(-1)补肺汤水煎剂,连续给药32 d。第33天开始将大鼠股动脉切开,放血处死,取右肺组织,用免疫组织化学S-P法检测TLR2、TLR4、My D88、核转录因子-κB(nuclear transcription-κB,NF-κB)蛋白基因的阳性表达,并用Western-Blot法检测肺组织中TLR2、My D88、NF-κB的含量。结果:与假手术组比较,模型组大鼠肺组织中NF-κB、TLR2、TLR4、My D88的表达,差异均有统计学意义(P<0.05);与模型组比较,补肺汤饮片汤剂组、西药组、补肺汤免煎颗粒剂高剂量组大鼠肺组织中NF-κB的表达,差异均有统计学意义(P<0.05);补肺汤饮片汤剂组、西药组、补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组大鼠肺组织中TLR2、TLR4的表达,差异均有统计学意义(P<0.05);补肺汤饮片汤剂组、补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组大鼠肺组织中My D88的表达,差异均有统计学意义(P<0.05)。与假手术组比较,模型组My D88、NF-κB、TLR2的含量差异均有统计学意义(P<0.05)。与模型组比较,西药组、补肺汤饮片汤剂组、补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组My D88、NF-κB的含量差异均有统计学意义(P<0.05);西药组、补肺汤免煎颗粒剂高剂量组、补肺汤免煎颗粒剂中剂量组TLR2的含量差异均有统计学意义(P<0.05)。结论:补肺汤免煎剂、汤剂均可同时抑制COPD大鼠肺组织中TLR2、TLR4两种Toll样受体的活性,通过My D88依赖性经典途径,调节下游NF-κB信号通路,从而改善气道重塑,但以TLR2通路为主,TLR4通路起协同作用,同时补肺汤通过调节TLR2/NF-κB信号通路改善COPD气道重塑的效果具有剂量依赖性。
Objective：To observe the influence of Lung-nourishing Decoction（ LND） on airway remodeling in stable chronic obstructive pulmonary disease（ COPD） rats,to find out whether it can regulate the immune function of COPD rats and inhibit airway inflammation through TLR2/NF-kappa B signal transduction pathway. Methods： Eigty-four rats were randomly divided into the sham operation group,the model group,the high dose group of LND free fried granules,the medium dose group of LND free fried granules,and the low dose group of LND free fried granules,the LND Decoction pieces group,the Western medicine group,with 12 rats in each group. Except the sham operation group,the other groups were made into COPD animal models by giving injection of lipopolysaccharide and smoking. The sham operation group was injected with the same amount of normal saline in the airway at the same time,and no passive smoking was used. After the success of establishing the model,the gavage was given on the second day,the sham operation group and the model group were given 10 m L ·（kg·d）^-1 physiological saline. The western medicine group was given 1 mg·（kg·d）^-1 montelukast sodium. The high dose group of LND free fried granules,the medium dose group of LND free fried granules,and the low dose group of LND free fried granules were given 27. 8 g·（kg·d）^-1,13. 9 g·（kg·d）^-1,and 7. 0 g·（kg·d）^-1 free fried decoction respectively. The LND Decoction pieces group was given 13. 9 g·（kg·d）^-1 Lungnourishing Decoction. Each group was treated for 32 days continuously. On the thirty-third day,the rats＇ femoral arteries were cut,and they were put to death by letting blood,and the right lung tissues were taken. The positive expressions of TLR2,TLR4,My D88,NF-kappa B protein gene were detected by immunohistochemical S-P method,and the contents of TLR2,My D88 and NFkappa B in lung tissues were detected by Western-Blot method. Results：Compared with sham operation group,the differences in the expressions of NF-kappa B,TLR2,TLR4 and My D88 in lung tissue of the model group were statistically different（ P〈0. 05）.Compared with the model group,all the differences in the expressions of NF-kappa B expression in the lung tissues of rats of the LND Decoction pieces group,the Western medicine group,and the high dose group of LND free fried granules were statistically significant（ P〈0. 05）. All the differences in the expressions of TLR2 and TLR4 in rats＇ lung tissues of the LND Decoction pieces group,the Western medicine group,the high dose group of LND free fried granules and the medium dose group of LND free fried granules were statistically significant（ P〈0. 05）. All the differences in the expressions of My D88 in rats＇ lung tissues of the LND Decoction pieces group,the high dose group of LND free fried granules and the medium dose group of LND free fried granules were statistically significant（ P〈0. 05）. Compared with the sham operation group,the differences of contents of My D88,NF-kappa B and TLR2 in the model group were statistically significant（ P〈0. 05）. Compared with the model group,there were significant differences in the contents of My D88 and NF-kappa B in the western medicine group,the LND Decoction pieces group,the high dose group of LND free fried granules and the medium dose group of LND free fried granules were statistically significant（P〈0. 05）. There were significant differences in the content of TLR2 between the western medicine group,the high dose group of LND free fried granules and the medium dose group of LND free fried granules. Conclusion： Both LND free fried granules and Lung-nourishing Decoction can inhibit the activity of two Toll-like receptors of TLR2 and TLR4 in the lung tissues of COPD rats.Through the My D88 dependent classical pathway,the downstream NF-kappa B signal pathway is regulated to improve the airway remodeling,but the TLR4 pathway plays a synergistic role in the TLR2 pathway. Meanwhile,Lung-nourishing Decoction improves the COPD airway remodeling by regulating the TLR2/NF-kappa B signaling pathway. The effect is dose-dependent.
作者
文秀华
王飞
苏凯
WEN Xiu-hua;WANG Fei;SU Kai(Sichuan second Chinese medicine hospital,Chengdu,Sichuan,China,610031;Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, China, 610075)
出处
《河南中医》
2018年第6期838-843,共6页
Henan Traditional Chinese Medicine
基金
四川省高等教育新世纪教育改革工程基金项目(编号:2015JY0267)
关键词
补肺汤
慢性阻塞性肺疾病
稳定期
转导通路
气道重塑
大鼠
Lung-nourishing Decoction
chronic obStructive pulmonary disease
stable phase
transduction pathway
airway remodeling
rats
