■哮喘,与炎症密切相关的慢性呼吸道疾病,提出了持续的治疗挑战。IALLIPF(le-Ala-Leu-Leu-Ile-Pro-Phe)是小米醇溶蛋白肽(MPP)之一,通过减少活性氧(ROS)的产生而显示出抗氧化生物活性。色氨酸(Trp,W)是已被证明具有抗炎作用的氨基酸。我们介绍了一种新型的组织蛋白酶B可激活的生物活性肽纳米载体,PEG-IALLIPF-GFLG-W(MPP-Trp),设计用于哮喘的免疫治疗。
■合成了MPP-Trp,纯化,通过动态光散射(DLS)和透射电子显微镜(TEM)研究了其特性。一氧化氮(NO)和促炎细胞因子(TNF-α,IL-6和IL-1β)进行检查以评估IALLIPF的抗炎作用,Trp和MPP-Trp.IALLIPF的免疫调节作用,通过流式细胞术研究Trp和MPP-Trp对Th1/Th2细胞群和细胞因子的影响,qRT-PCR和ELISA测定。我们通过肺组织学和ELISA分析探讨MPP-Trp在哮喘小鼠模型中的治疗作用。有必要通过CCK8测定和苏木精和伊红(HE)染色进行组织病理学分析来研究MPP-Trp的生物相容性。
■在哮喘外周血单核细胞(PBMC)中,IALLIPF,Trp和MPP-Trp能够通过抑制一氧化氮(NO)和促炎细胞因子(TNF-α,IL-6和IL-1β),尤其是MPP-Trp.MPP-Trp显著上调Th1细胞水平,同时显著降低Th2细胞水平。此外,MPP-Trp有效提高干扰素-γ(IFN-γ)的表达和产生,来自Th1细胞的必需细胞因子。此外,MPP-Trp显著降低哮喘发病关键细胞因子的mRNA表达和水平,如白细胞介素-4(IL-4),白细胞介素-13(IL-13),和白细胞介素-5(IL-5),在哮喘PBMC中。MPP-Trp改善哮喘小鼠肺部病理改变并显著抑制OVA诱导的炎症。它对用各种浓度或持续时间的MPP-Trp处理的哮喘-PBMC中的细胞活力几乎没有影响。无病理变化,包括在心里,肝脏,脾,脾肺,和肾脏组织,在用MPP-Trp(20mg/kg)处理的非致敏和非攻击小鼠中观察到。
■我们的研究表明,MPP-Trp对Th1/Th2细胞群具有免疫调节作用,治疗哮喘至关重要。它通过将免疫应答转向Th1-显性模式显著缓解OVA诱导的哮喘,从而减少Th2驱动的炎症。因此,这种新型的生物活性肽纳米载体,MPP-Trp,有望成为哮喘免疫治疗的候选药物。
UNASSIGNED: Asthma, a chronic respiratory disease closely associated with inflammation, presents ongoing treatment challenges. IALLIPF (le-Ala-Leu-Leu-Ile-Pro-Phe) is one of millet prolamins peptides (MPP) which shows anti-oxidant bioactivity by reducing the production of reactive oxygen species (ROS). Tryptophan (Trp, W) is an amino acid that has been demonstrated to possess anti-inflammatory effects. We introduce a novel cathepsin B-activatable bioactive peptides nanocarrier, PEG-IALLIPF-GFLG-W (MPP-Trp), designed for immunotherapy of asthma.
UNASSIGNED: MPP-Trp is synthesized, purified, and its characteristics are investigated by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The yield of nitric oxide (NO) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) are examined to evaluate anti-inflammatory effects of IALLIPF, Trp and MPP-Trp. The
immunomodulatory effects of IALLIPF, Trp and MPP-Trp on Th1/Th2 cell populations and cytokines are investigated by flow cytometry, qRT-PCR and ELISA assays. We explore the therapeutic effect of MPP-Trp in the mouse model of asthma by the analysis of lung histology and ELISA. It is necessary to study the biocompatibility of MPP-Trp by CCK8 assay and histopathologic analysis using hematoxylin and eosin (HE) staining.
UNASSIGNED: In asthmatic peripheral blood mononuclear cells (PBMCs), IALLIPF, Trp and MPP-Trp are able to significantly alleviate inflammation by inhibiting the yield of nitric oxide (NO) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β), especially MPP-Trp. MPP-Trp significantly upregulates Th1 cell levels while notably reducing Th2 cell levels. Furthermore, MPP-Trp effectively elevates the expression and production of interferon-gamma (IFN-γ), an essential cytokine from Th1 cells. Additionally, MPP-Trp markedly diminishes the mRNA expression and levels of key asthma pathogenesis cytokines, such as interleukin-4 (IL-4), interleukin-13 (IL-13), and interleukin-5 (IL-5), in asthma PBMCs. MPP-Trp ameliorates pulmonary pathological alterations and significantly inhibits OVA-induced inflammation in mice with asthma. It has little influence on the cell viability in Asthma-PBMCs treated with various concentrations or durations of MPP-Trp. No pathological changes, including in the heart, liver, spleen, lung, and kidney tissues, are observed in non-sensitized and non-challenged mice treated with MPP-Trp (20 mg/kg).
UNASSIGNED: Our research demonstrates that MPP-Trp has
immunomodulatory effects on Th1/Th2 cell populations, essential in managing asthma. It considerably alleviates OVA-induced asthma by shifting the immune response towards a Th1-dominant profile, thereby reducing Th2-driven inflammation. Therefore, this novel bioactive peptide nanocarrier, MPP-Trp, holds promise as a candidate for asthma immunotherapy.