Li Pan-Ying, Yuan Pei-Pei, Hou Ying, Gao Li-Yuan, Wei Ya-Xin, Ruan Yuan, Chen Yi, Fu Yang, Zheng Xiao-Ke, Feng Wei-Sheng
School of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China.
School of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China Henan Engineering Technology Research Center for Traditional Chinese Medicine Development Zhengzhou 450046, China Co-construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan & Education Ministry of China Zhengzhou 450046, China.
Zhongguo Zhong Yao Za Zhi. 2022 Feb;47(4):1009-1016. doi: 10.19540/j.cnki.cjcmm.20211103.705.
The present study investigated the effect of active components of Descurainia sophia on allergic asthma and explored the underlying mechanism. SD male rats were randomly divided into a normal group(NC), a model group(M), a D. sophia decoction group(DS), a D. sophia fatty oil group(FO), a D. sophia flavonoid glycoside group(FG), a D. sophia oligosaccharide group(Oli), and a positive drug dexamethasone group(Y). The allergic asthma model was induced in rats by intraperitoneal injection of ovalbumin(OVA) and aluminum hydroxide gel adjuvant(sensitization) and atomization of OVA solution(excitation). After modeling, asthma-related indicators, tracheal phenol red excretion, inflammatory cell levels in the peripheral blood, lung permeability index(LPI), and oxygenation index(OI) of rats were detected. The pathological changes of lung tissues were observed by HE staining. Enzyme-linked immunosorbent assay(ELISA) was used to detect the content of inflammatory factors immunoglobulin E(IgE), interleukin-4(IL-4), and interferon-γ(IFN-γ) in the bronchoalveolar lavage fluid(BALF) and the content of endothelin-1(ET-1) and angiotensin-converting enzyme(ACE) in lung tissue homogenate. The serum content of nitric oxide(NO) was detected by colorimetry. Western blot was employed to determine the protein expression of Toll-like receptor 4(TLR4), nuclear factor κB-p65(NF-κB-p65), phosphorylated NF-κB-p65(p-NF-κB-p65), myosin light chain kinase(MLCK), vascular endothelial cadherin(VE cadherin), connexin 43, and claudin 5, and the mechanism of active components of D. sophia on allergic asthma was explored. As revealed by the results, the M group showed extensive infiltration of inflammatory cells around the bronchus of the lung tissues of the allergic asthma rats, thickened bronchial wall, severely deformed alveolar structure, increased number of wheezes, the content of IgE, IL-4, ET-1, and ACE, inflammatory cells, and LPI, and reduced latency of asthma, tracheal phenol red excretion, IFN-γ, NO content, and OI. After the intervention of the active components of D. sophia, the DS, FO, FG, Oli, and Y groups showed improved asthma-related indicators, tracheal phenol red excretion, and lung tissue lesions in allergic asthma rats, and the effects in the FO and Oli groups were superior. The content of inflammatory factors in BALF was recovered in the DS, FO, and Y groups and the FG and Oli groups. The number of inflammatory cells in rats was reduced in the DS and FO groups, and the FG, Oli, and Y groups to varying degrees, and the effect in the FO group was superior. DS, FO, Oli, and Y reduced ET-1, ACE, and LPI and increased NO and OI. FG recovered NO, ET-1, ACE, LPI, and OI to improve lung epithelial damage and permeability. Further investigation of inflammation-related TLR4/NF-κB pathways, MLCK, and related skeleton protein levels showed that TLR4, NF-κB-p65, p-NF-κB-p65, and MLCK levels were increased, and VE cadherin, connexin 43, and claudin 5 were reduced in the M group. DS, FO, FG, Oli, and Y could reduce the protein expression related to the TLR4 pathway to varying degrees, and regulate the protein expression of MLCK, VE cadherin, connexin 43, and claudin 5. It is inferred that the active components of D. sophia improve lung permeability in rats with allergic asthma presumedly by regulating the TLR4/NF-κB signaling pathway to improve airway inflammation, mediating MLCK and connexin, and regulating epithelial damage.
本研究探讨了播娘蒿活性成分对过敏性哮喘的影响,并探究其潜在机制。将雄性SD大鼠随机分为正常组(NC)、模型组(M)、播娘蒿水煎剂组(DS)、播娘蒿脂肪油组(FO)、播娘蒿黄酮糖苷组(FG)、播娘蒿低聚糖组(Oli)和阳性药物地塞米松组(Y)。通过腹腔注射卵清蛋白(OVA)和氢氧化铝凝胶佐剂(致敏)以及雾化OVA溶液(激发)诱导大鼠过敏性哮喘模型。造模后,检测大鼠哮喘相关指标、气管酚红排泄量、外周血炎症细胞水平、肺通透性指数(LPI)和氧合指数(OI)。通过苏木精-伊红(HE)染色观察肺组织的病理变化。采用酶联免疫吸附测定(ELISA)法检测支气管肺泡灌洗液(BALF)中炎症因子免疫球蛋白E(IgE)、白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)的含量以及肺组织匀浆中内皮素-1(ET-1)和血管紧张素转换酶(ACE)的含量。采用比色法检测血清一氧化氮(NO)含量。采用蛋白质免疫印迹法检测Toll样受体4(TLR4)、核因子κB-p65(NF-κB-p65)、磷酸化NF-κB-p65(p-NF-κB-p65)、肌球蛋白轻链激酶(MLCK)、血管内皮钙黏蛋白(VE钙黏蛋白)、连接蛋白43和闭合蛋白5的蛋白表达,探究播娘蒿活性成分对过敏性哮喘的作用机制。结果显示,M组过敏性哮喘大鼠肺组织支气管周围炎症细胞广泛浸润,支气管壁增厚,肺泡结构严重变形,喘息次数增加,IgE、IL-4、ET-1和ACE含量、炎症细胞及LPI升高,哮喘潜伏期缩短,气管酚红排泄量、IFN-γ、NO含量及OI降低。播娘蒿活性成分干预后,DS、FO、FG、Oli和Y组过敏性哮喘大鼠哮喘相关指标、气管酚红排泄量及肺组织病变均有改善,其中FO和Oli组效果更佳。DS、FO和Y组以及FG和Oli组BALF中炎症因子含量均有所恢复。DS和FO组以及FG、Oli和Y组大鼠炎症细胞数量不同程度减少,其中FO组效果更佳。DS、FO、Oli和Y组降低了ET-1、ACE和LPI,升高了NO和OI。FG组恢复了NO、ET-1、ACE、LPI和OI,改善了肺上皮损伤和通透性。进一步研究炎症相关的TLR4/NF-κB信号通路、MLCK及相关骨架蛋白水平发现,M组TLR4、NF-κB-p65、p-NF-κB-p65和MLCK水平升高,VE钙黏蛋白、连接蛋白43和闭合蛋白5降低。DS、FO、FG、Oli和Y组可不同程度降低与TLR4通路相关的蛋白表达,调节MLCK、VE钙黏蛋白、连接蛋白43和闭合蛋白5的蛋白表达。推测播娘蒿活性成分可能通过调节TLR4/NF-κB信号通路改善气道炎症,介导MLCK和连接蛋白,调节上皮损伤,从而改善过敏性哮喘大鼠的肺通透性。