基于单宁酸的ROS响应型水凝胶在巨噬细胞炎症中的抗炎抗氧化作用OA

Objective·To develop a reactive oxygen species(ROS)-responsive hydrogel based on tannic acid(TA),and to evaluate its anti-inflammatory and antioxidant effects in an lipopolysaccharide(LPS)-induced RAW264.7 macrophage inflammation model.Methods·Phenylboronic acid-modified hyaluronic acid(HP)was complexed with TA to form the HP-TA hydrogel,and its ROS-responsive properties were assessed by using a hydrogen peroxide consumption assay.The biocompatibility of HP and TA in RAW264.7 macrophages was evaluated by cell counting kit-8(CCK-8)assay,cell cycle assay,and live/dead staining.An LPS-induced inflammation model of RAW264.7 macrophages was established and treated with the HP-TA hydrogel.The mRNA and protein expression levels of pro-inflammatory genes,including inducible nitric oxide synthase(Inos)and tumor necrosis factor-α(Tnf-α),as well as antioxidant genes,including NAD(P)H:quinone oxidoreductase 1(Nqo1)and heme oxygenase-1(Ho-1),were analyzed by quantitative real-time PCR(qPCR)and Western blotting,respectively.A 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescent probe combined with flow cytometry and nitric oxide content detection was performed to analyze the total ROS-scavenging capacity of the hydrogel.Results·The HP-TA hydrogel was successfully synthesized and confirmed to be ROS-responsive.At safe concentrations(200 μg/mL for HP and 25 μg/mL for TA),both materials exhibited good biocompatibility with RAW264.7 macrophages.The HP-TA hydrogel significantly inhibited the mRNA and protein expression of the pro-inflammatory genes Inos and Tnf-α,and promoted the mRNA and protein expression of the antioxidant genes Nqo1 and Ho-1 in inflammatory macrophages(all P<0.05).In addition,the HP-TA hydrogel significantly reduced the total intracellular ROS level(P<0.001).Conclusion·The HP-TA hydrogel can effectively suppress LPS-induced inflammatory responses in macrophages,exert antioxidant effects by scavenging ROS,and upregulate the expression of antioxidant pathway proteins.This study reveals the potential of the HP-TA hydrogel as a ROS-responsive intelligent drug delivery system.