| 刘斐,虞坚尔,薛征,等.平喘方对哮喘模型小鼠pDC/cDC通路的干预作用研究[J].浙江中医药大学学报,2019,43(7):693-701. |
| 平喘方对哮喘模型小鼠pDC/cDC通路的干预作用研究 |
| Intervention Role of Pingchuan Recipe on PDC/cDC in Lung Tissue of Mice with Asthma |
| DOI：10.16466/j.issn1005-5509.2019.07.017 |
| 中文关键词: 支气管哮喘 平喘方 pDC/cDC T细胞，气道炎症 气道重建 |
| 英文关键词: bronchial asthma Pingchuan Recipe pDC/cDC T cell airway inflammation airway remodeling |
| 基金项目:无锡市卫计委青年医学人才项目(QNRC037)；国家自然科学基金面上项目(81574026) |
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| 中文摘要: |
| [目的]通过观察平喘方对支气管哮喘模型小鼠肺组织浆细胞样树突状细胞(plasmacytoid dendritic cell，pDC)/传统DC(conventional DC，cDC)通路各因子的影响，阐释平喘方治疗哮喘的机制及药物间的相互作用。[方法]将40只雄性BALB/c小鼠随机分为空白组、模型组、地米组和平喘方组，每组10只。采用10%鸡卵白蛋白(ovalbumin，OVA)致敏液腹腔注射致敏和5%OVA雾化吸入激发，建立BALB/c小鼠支气管哮喘模型。小鼠激发开始时按20mL/kg·d灌胃相应的药物，用药1周后处死小鼠，采集支气管肺泡灌洗液，以ELISA法检测支气管肺泡灌洗液中白细胞介素-6(interleukin-6，IL-6)、白细胞介素-17(interleukin-17，IL-17)、白细胞介素-23(interleukin-23，IL-23)及转化生长因子-β(transforming growth factor-β，TGF-β)的含量；分离小鼠肺组织，HE染色后镜下观察肺组织形态，Western blot检测肺组织中维甲酸相关孤儿受体(retinoid-related orphan nuclear receptor，RORγt)及叉头蛋白P3(forkhead box P3，FOXP3)的蛋白表达量；Real-time PCR测定肺组织中吲哚胺-2,3-二氧化酶(indoleamine-2,3-dioxygenase，IDO)、肿瘤坏死因子超家族4(tumor necrosis factor superfamily4，TNFSF4)的基因相对表达量；Masson染色检测肺组织胶原蛋白面积、气道平滑肌面积(airway smooth muscle area，WAm)、气道内壁面积(airway inner wall area，WAi)。[结果]与空白组比较，模型组支气管肺泡灌洗液中IL-6、IL-17、IL-23、TGF-β的含量以及肺组织中RORγt、TNFSF4表达均增高，FOXP3、IDO表达均降低，胶原蛋白面积、WAm、WAi均增加，差异有统计学意义(P＜0.05)。与模型组比较，地米组、平喘方组病理切片表现均有所改善，支气管肺泡灌洗液中IL-6、IL-17、IL-23、TGF-β的含量、肺组织中RORγt、TNFSF4表达均减低，FOXP3、IDO表达均增高，差异有统计学意义(P＜0.05)；与地米组比较，平喘方组IL-17的含量、RORγt、TNFSF4、FOXP3、IDO的表达量及胶原蛋白面积、WAm、WAi差异无统计学意义(P＞0.05)。[结论]平喘方对pDC/cDC的干预可能是诱导气道免疫耐受的一个切入点，从而抑制T细胞表达失衡，改善气道炎症及气道重建。 |
| 英文摘要: |
| [Objective] To study Pingchuan Recipe on plasmacytoid dendritic cell(pDC)/conventional DC(cDC) in lung tissue of mice with asthma to explain its mechanism and compatibility of medicines.[Methods]Forty male BALB/c mice were randomly divided into blank group, model group, dexamethasone group, Pingchuan Recipe group, 10 in each group. 10% ovalbumin(OVA) was injected intraperitoneally and 5% OVA atomization inhalation challenge, established the BALB/c mouse bronchial asthma model. Gavaged the appropriate drugs of 20mL/kg·d from the beginning of stimulation. The mouse lung tissue collected after 1 week. The contents of interleukin-6(IL-6), interleukin-17(IL-17), interleukin-23(IL-23) and transforming growth factor-β(TGF-β) in branchoalveolar lavage fluid(BALF) were detected by ELISA. Lung tissues of mice were extracted and observed by HE staining. The expression of retinoid-related orphan nuclear receptor(RORγt) and forkhead box P3(FOXP3) in lung tissue was determined by Western blot. Indoleamine-2,3-dioxygenase(IDO) and tumor necrosis factor superfamily 4(TNFSF4) were determined by Real-time PCR. Collagen area, airway smooth muscle area (WAm) and airway inner wall area(WAi) in lung tissue were detected by Masson staining.[Results] Compared with blank group, the contents of IL-6, IL-17, IL-23, TGF-β in BALF, the expression of RORγt and TNFSF4 in lung tissue of model group increased, while the expression of FOXP3 and IDO decreased, and collagen area, WAm and WAi increased(P＜0.05).Compared with model group, the pathological slices of dexamethasone group and Pingchuan Recipe group were improved, the contents of IL-6, IL-17, IL-23, TGF-β in BALF, the expression of RORγt and TNFSF4 in lung tissue decreased(P＜0.05), the expression of FOXP3 and IDO increased with statistical significance(P＜0.05). Compared with dexamethasone group, there were no significant difference of the expression of IL-17, RORγt, TNFSF4, FOXP3, IDO and collagen area, WAm and WAi in Pingchuan Recipe group(P＞0.05). [Conclusion] The intervention of Pingchuan Recipe on pDC/cDC may be a breakthrough point to form airway immune tolerance, so as to suppress the imbalance expression of T cells and improve airway inflammation and remodeling. |
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