Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, People's Republic of China.
Int J Nanomedicine. 2022 Oct 17;17:4843-4860. doi: 10.2147/IJN.S378073. eCollection 2022.
Oral administration of proteins/peptides is challenging in clinical application due to their instability and susceptibility in the gastrointestinal tract.
The in situ polymerization on the surface of enzymes was used to encapsulate antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)) in polymeric shells, and the reactive oxygen species (ROS) scavenging ability was monitored based on DCFH-DA probe using flow cytometry and confocal laser scanning microscopy. The mRNA expression level of pro-inflammatory factors was assessed by real-time qPCR, using lipopolysaccharide-induced RAW264.7 cells as a model. Finally, the enzyme capsules were orally administered for the treatment of inflammatory bowel disease using dextran sodium sulfate (DSS)-induced colitis mice as a model, based on the evaluation of the disease-associated index, ROS level and pro-inflammatory cytokines' expression.
The enzyme capsules could effectively scavenge the intracellular reactive oxygen species (ROS) through the cascade catalysis of SOD and CAT, and thus protect the cells from ROS-induced oxidative damage. Meanwhile, the enzyme capsules could inhibit the secretion of pro-inflammatory cytokines from macrophages, thereby achieving favorable anti-inflammation effect. Oral administration of enzyme capsules could facilitate the accumulation of enzymes in the inflamed colon tissues of DSS-induced colitis mice. Moreover, the oral delivery of enzyme capsules could effectively alleviate the symptoms associated with colitis, attributing to the excellent ROS scavenging ability and the inhibition of pro-inflammatory cytokines' level.
In summary, our findings provided a promising approach to construct enzyme-based nano-formulations with favorable therapeutic efficacy and biocompatibility, exhibiting great potential in the treatment of gastrointestinal diseases in an oral administration manner.
由于蛋白质/肽在胃肠道中不稳定且易被降解，因此经口给药在临床应用中具有挑战性。
采用酶表面原位聚合的方法，将抗氧化酶（超氧化物歧化酶（SOD）和过氧化氢酶（CAT））包埋在聚合物壳中，并用 DCFH-DA 探针通过流式细胞术和共聚焦激光扫描显微镜监测其活性氧（ROS）清除能力。采用实时 qPCR 评估脂多糖诱导的 RAW264.7 细胞模型中促炎因子的 mRNA 表达水平。最后，以葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠为模型，通过评估疾病相关指标、ROS 水平和促炎细胞因子的表达，口服给予酶胶囊治疗炎症性肠病。
酶胶囊可以通过 SOD 和 CAT 的级联催化有效地清除细胞内的活性氧（ROS），从而保护细胞免受 ROS 诱导的氧化损伤。同时，酶胶囊可以抑制巨噬细胞中促炎细胞因子的分泌，从而发挥良好的抗炎作用。酶胶囊的口服给药可以促进酶在 DSS 诱导的结肠炎小鼠炎症结肠组织中的积累。此外，酶胶囊的口服递送可以有效缓解与结肠炎相关的症状，这归因于其出色的 ROS 清除能力和对促炎细胞因子水平的抑制作用。
总之，我们的研究结果为构建具有良好治疗效果和生物相容性的基于酶的纳米制剂提供了一种有前景的方法，有望通过口服给药的方式治疗胃肠道疾病。
