人β-防御素-3调控呼吸道合胞病毒致哮喘幼鼠的气道重塑及白细胞激活-东南大学学报医学版

人β-防御素-3调控呼吸道合胞病毒致哮喘幼鼠的气道重塑及白细胞激活

单位：海南医学院第一附属医院儿科, 海南海口 570100

分类号：R562.25;R-33

出版年·卷·期（页码）：2022·41·第二期（183-192）

摘要：

目的:探究人β-防御素-3(hBD3)对呼吸道合胞病毒(RSV)感染诱发哮喘幼鼠气道重塑与白细胞激活的影响及其机制。方法:采用卵清白蛋白(OVA)合并RSV诱发建立幼鼠哮喘模型,期间予100 μg·kg^-1·d^-1 hBD3溶液灌胃;制备肺泡灌洗液(BALF),瑞氏染色测定BALF细胞总数与各细胞计数,ELISA法检测BALF和血清中免疫球蛋白IgE、白细胞介素(IL)-4、IL-5、肿瘤坏死因子-α(TNF-α)水平;HE染色、PAS染色和Masson染色观察肺组织病理学变化情况,观察切片测定各项气道重塑指标,免疫组化染色检测肺组织α-平滑肌肌动蛋白(α-SMA)表达,蛋白质印迹法检测肺组织生长因子与自噬相关蛋白的表达。结果:相较于模型组,经hBD3作用的幼鼠BALF中细胞总数以及嗜酸粒细胞、中性粒细胞、淋巴细胞的比例均降低,BALF和血清中IgE、IL-4、IL-5、TNF-α水平均下降,肺组织损伤明显缓解,炎症细胞浸润减轻,杯状细胞减少,肺组织胶原纤维面积较小,管壁面积/气管内周长(WAt/Pi)、支气管平滑肌面积/气管内周长(WAm/Pi)及支气管平滑肌细胞核数目/气管内周长(N/Pi)均下降,α-SMA阳性表达率下降,同时肺组织中血管内皮生长因子(VEGF)、转化生长因子-β1(TGF-β1)、胰岛素样生长因子-1(IGF-1)以及自噬相关蛋白LC3-Ⅱ与Beclin-1的蛋白表达水平均下调,P62蛋白表达水平则上调,差异具有统计学意义(P＜0.05)。结论:hBD3能够抑制RSV感染诱发哮喘幼鼠模型的气道炎症和气道重塑过程,介导白细胞失活,该作用可能与调控自噬水平相关。

Objective: To explore the effect and mechanism of human β-defensin-3(hBD3) on airway remodeling and white blood cell activation in young asthmatic rats induced by respiratory syncytial virus(RSV) infection. Methods: Ovalbumin(OVA) combined with RSV was used to induce an asthma model in young rats, during which 100 μg·kg^-1·d^-1 hBD3 solution was given by gavage. Lavage fluid(BALF) was prepared, and the total number of BALF cells and cell counts were determined by Wright's staining. The levels of immunoglobulin IgE, interleukin-4(IL-4), IL-5, and tumor necrosis factor-α(TNF-α) in BALF and serum were detected by ELISA. HE staining, PAS staining and Masson staining were used to observe the pathological changes of lung tissue and various airway remodeling indexes observed by pathological sections. Immunohistochemical staining and Western blot were used to detect the lung tissue α-SMA expression and the expression of growth factors and autophagy-related proteins in lung tissue leukocytes respectively. Results: Compared with the model group, the total number of cells in BALF, the proportion of eosinophils, neutrophils and lymphocytes in the pups treated with hBD3 were reduced. The levels of IgE, IL-4, IL-5 and TNF-α in BALF and serum decreased, and the lung tissue damage was significantly alleviated. Inflammatory cell infiltration and goblet cells were reduced. The area of collagen fibers in lung tissue was smaller, WAt/Pi, WAm/Pi and N/Pi were all reduced, and the positive expression rate of α-SMA was decreased. At the same time, the expression protein levels of VEGF, TGF-β1, IGF-1, LC3-Ⅱ and Beclin-1 in the tissues were down-regulated and the expression protein levels of P62 were up-regulated, the difference being statistically significant(P＜0.05). Conclusion: hBD3 can inhibit the airway inflammation and airway remodeling process in young rats with asthma induced by RSV infection, and mediate leukocyte inactivation, which may be related to the regulation of autophagy.

参考文献：

[1] QUINN L A,SHIELDS M D,SINDA I,et al.Respiratory syncytial virus prophylaxis for prevention of recurrent childhood wheeze and asthma:a systematic review[J].Syst Rev,2020,25,9(1):269.
[2] SHI T,OOI Y,ZAW E M,et al.Association between respiratory syncytial virus-associated acute lower respiratory infection in early life and recurrent wheeze and asthma in later childhood[J].J Infect Dis,2020,222(Suppl 7):S628-S633.
[3] ZHOU Y,TONG L,LI M,et al.Recurrent wheezing and asthma after respiratory syncytial virus bronchiolitis[J].Front Pediatr,2021,9:649003.
[4] MUCKE P A,OSTRZINSKI A,HAMMERSCHMIDT S,et al.Proteomic adaptation of streptococcus pneumoniae to the antimicrobial peptide human beta defensin 3(hBD3) in comparison to other cell surface stresses[J].Microorganisms,2020,8(11):1697.
[5] LI L,BIAN T,LYU J,et al.Human β-defensin-3 alleviates the progression of atherosclerosis accelerated by Porphyromonas gingivalis lipopolysaccharide[J].Int Immunopharmacol,2016,38:204-213.
[6] LYU J,BIAN T,CHEN B,et al.β-defensin 3 modulates macrophage activation and orientation during acute inflammatory response to Porphyromonas gingivalis lipopolysaccharide[J].Cytokine,2017,92:48-54.
[7] PARK O J,KIM J,AHN K B,et al.A 15-amino acid C-terminal peptide of beta-defensin-3 inhibits bone resorption by inhibiting the osteoclast differentiation and disrupting podosome belt formation[J].J Mol Med(Berl),2017,95(12):1315-1325.
[8] 蒋雄斌,朱毅,殷凯生.用呼吸道合胞病毒诱导致敏小鼠建立重度支气管哮喘动物模型[J].中华结核和呼吸杂志,2006,29(5):344-345.
[9] CHEN L,LV Z,GAO Z,et al.Human β-defensin-3 reduces excessive autophagy in intestinal epithelial cells and in experimental necrotizing enterocolitis[J].Sci Rep,2019,9(1):19890.
[10] BIAN T,LI H,ZHOU Q,et al.Human β-defensin 3 reduces TNF-α-induced inflammation and monocyte adhesion in human umbilical vein endothelial cells[J].Mediators Inflamm,2017,2017:8529542.
[11] CUI D,LYU J,LI H,et al.Human β-defensin 3 inhibits periodontitis development by suppressing inflammatory responses in macrophages[J].Mol Immunol,2017,91:65-74.
[12] FEHRENBACH H,WAGNER C,WEGMANN M.Airway remodeling in asthma:what really matters[J].Cell Tissue Res,2017,367(3):551-569.
[13] BOULET L P.Airway remodeling in asthma:update on mechanisms and therapeutic approaches[J].Curr Opin Pulm Med,2018,24(1):56-62.
[14] LEE C G,LINK H,BALUK P,et al.Vascular endothelial growth factor(VEGF) induces remodeling and enhances TH2-mediated sensitization and inflammation in the lung[J].Nat Med,2004,10(10):1095-1103.
[15] ITO I,FIXMAN ED,ASAI K,et al.Platelet-derived growth factor and transforming growth factor-beta modulate the expression of matrix metalloproteinases and migratory function of human airway smooth muscle cells[J].Clin Exp Allergy,2009,39(9):1370-1380.
[16] HAN Y Y,YAN Q,CHEN W,et al.Serum insulin-like growth factor-1,asthma,and lung function among British adults[J].Ann Allergy Asthma Immunol,2021,126(3):284-291.
[17] ROSSAINT J,MARGRAF A,ZARBOCK A.Role of platelets in leukocyte recruitment and resolution of inflammation[J].Front Immunol,2018,9:2712.
[18] BAN G Y,PHAM D L,TRINH T H,et al.Autophagy mechanisms in sputum and peripheral blood cells of patients with severe asthma:A new therapeutic target[J].Clin Exp Allergy,2016,46(1):48-59.
[19] KIM S R,SONG J H,AHN J H,et al.Antiviral and anti-inflammatory activity of budesonide against human rhinovirus infection mediated via autophagy activation[J].Antiviral Res,2018,151(10):87-96.

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