Recombinant adeno-associated virus carrying thymosin β(4) suppresses experimental colitis in mice.
Recombinant adeno-associated virus carrying thymosin β(4) suppresses experimental colitis in mice.
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DOI：
10.3748/wjg.v23.i2.242
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发表时间：
2017-01-14
影响因子：
4.3
通讯作者：
Hao ZM
中科院分区：
文献类型：
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作者：
Zheng XY;Lv YF;Li S;Li Q;Zhang QN;Zhang XT;Hao ZM
目的探讨携带胸腺素β4的重组腺相关病毒（AAV-Tβ4）结肠内给药对小鼠结肠炎的保护作用。制备了T β4腺相关病毒载体，并将其用于小鼠结肠内，以介导Tβ4的分泌表达。用右旋糖酐硫酸钠（DSS）诱导小鼠溃疡性结肠炎，用2，4，6-三硝基苯磺酸（TNBS）建立类似克罗恩病的小鼠结肠炎模型。观察各组大鼠结肠病变程度及结肠损伤程度，并进行分级，以评价AAV-Tβ4对结肠炎的治疗作用。采用生化分析法测定髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量。采用ELISA法检测结肠组织中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-1 β）和IL-10的水平，TUNEL法检测结肠黏膜上皮细胞凋亡，免疫组化法检测结肠黏膜上皮细胞增殖。重组腺相关病毒可将LacZ和Tβ4有效地递送到小鼠结肠组织中，并且AAV-Tβ4可导致Tβ4在小鼠结肠中的强表达。在DSS和TNBS结肠炎模型中，AAV-Tβ4处理的小鼠显示出明显减轻的结肠损伤和降低的结肠粘膜上皮细胞凋亡率。AAV-Tβ4显著减少炎症细胞浸润并减轻小鼠炎症结肠中的氧化应激，表现为MPO活性和MDA含量降低以及SOD活性升高。AAV-Tβ4还调节结肠TNF-α、IL-1β和IL-10水平，并抑制DSS和TNBS处理小鼠结肠上皮细胞的代偿性增殖。Tβ4对小鼠结肠炎具有保护作用，提示AAV-Tβ4有可能成为一种治疗炎症性肠病的药物。
To investigate the protective effect of a recombinant adeno-associated virus carrying thymosin β4 (AAV-Tβ4) on murine colitis via intracolonic administration. AAV-Tβ4 was prepared and intracolonically used to mediate the secretory expression of Tβ4 in mouse colons. Dextran sulfate sodium (DSS) was applied to induce the murine ulcerative colitis, and 2,4,6-trinitrobenzene sulfonic acid (TNBS) was used to establish a mouse colitis model resembling Crohn’s disease. The disease severity and colon injuries were observed and graded to reveal the effects of AAV-Tβ4 on colitis. The activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were determined using biochemical assays. Colonic levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-10 were measured using ELISA, and mucosal epithelial cell apoptosis and proliferation were detected by TUNEL assay and immunochemistry, respectively. Recombinant AAVs efficiently delivered LacZ and Tβ4 into the colonic tissues of the mice, and AAV-Tβ4 led to a strong expression of Tβ4 in mouse colons. In both the DSS and TNBS colitis models, AAV-Tβ4-treated mice displayed distinctly attenuated colon injuries and reduced apoptosis rate of colonic mucosal epithelia. AAV-Tβ4 significantly reduced inflammatory cell infiltrations and relieved oxidative stress in the inflamed colons of the mice, as evidenced by decreases in MPO activity and MDA content and increases in SOD activity. AAV-Tβ4 also modulated colonic TNF-α, IL-1β and IL-10 levels and suppressed the compensatory proliferation of colonic epithelial cells in DSS- and TNBS-treated mice. Tβ4 exerts a protective effect on murine colitis, indicating that AAV-Tβ4 could potentially be developed into a promising agent for the therapy of inflammatory bowel diseases.