Qu Jie, Pei Hui, Li Xin-Ze, Li Yan, Chen Jian-Ming, Zhang Min, Lu Zhong-Qiu
Emergency Department, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Wenzhou Key Laboratory of Emergency and Disaster Medicine, Wenzhou, China.
Front Pharmacol. 2024 Jul 9;15:1414918. doi: 10.3389/fphar.2024.1414918. eCollection 2024.
Diquat (DQ) poisoning can cause multiple organ damage, and the kidney is considered to be the main target organ. Increasing evidence shows that alleviating oxidative stress and inflammatory response has promising application prospects. Epigallocatechin gallate (EGCG) has potent antioxidant and anti-inflammatory effects. In this study, red blood cell membrane (RBCm)-camouflaged polylactic-co-glycolic acid (PLGA) nanoparticles (NPs) were synthesized to deliver EGCG (EGCG-RBCm/NPs) for renal injury induced by DQ. Human renal tubular epithelial cells (HK-2 cells) were stimulated with 600 μM DQ for 12 h and mice were intraperitoneally injected with 50 mg/kg b.w. DQ, followed by 20 mg/kg b.w./day EGCG or EGCG-RBCM/NPs for 3 days. The assessment of cellular vitality was carried out using the CCK-8 assay, while the quantification of reactive oxygen species (ROS) was performed through ROS specific probes. Apoptosis analysis was conducted by both flow cytometry and TUNEL staining methods. Pathological changes in renal tissue were observed. The expressions of NLRP3, IL-1β, IL-18, NFκB and Caspase1 were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunohistochemistry, immunofluorescence, and Western blot. The results showed that the DQ group had increased ROS expression, increased the level of oxidative stress, and increased apoptosis rate compared with the control group. Histopathological analysis of mice in the DQ group showed renal tubular injury and elevated levels of blood urea nitrogen (BUN), serum creatinine (SCr), kidney injury molecule-1 (KIM-1), and cystatin C (Cys C). Furthermore, the DQ group exhibited heightened expression of NLRP3, p-NFκB p65, Caspase1 p20, IL-1β, and IL-18. However, EGCG-RBCm/NPs treatment mitigated DQ-induced increases in ROS, apoptosis, and oxidative stress, as well as renal toxicity and decreases in renal biomarker levels. Meanwhile, the expression of the above proteins were significantly decreased, and the survival rate of mice was ultimately improved, with an effect better than that of the EGCG treatment group. In conclusion, EGCG-RBCm/NPs can improve oxidative stress, inflammation, and apoptosis induced by DQ. This effect is related to the NF-κB/NLRP3 inflammasome pathway. Overall, this study provides a new approach for treating renal injury induced by DQ.
敌草快（DQ）中毒可导致多器官损伤，肾脏被认为是主要靶器官。越来越多的证据表明，减轻氧化应激和炎症反应具有广阔的应用前景。表没食子儿茶素没食子酸酯（EGCG）具有强大的抗氧化和抗炎作用。在本研究中，合成了红细胞膜（RBCm）伪装的聚乳酸-乙醇酸共聚物（PLGA）纳米颗粒（NPs）来递送EGCG（EGCG-RBCm/NPs），用于治疗DQ诱导的肾损伤。用600 μM DQ刺激人肾小管上皮细胞（HK-2细胞）12小时，给小鼠腹腔注射50 mg/kg体重的DQ，随后每天注射20 mg/kg体重的EGCG或EGCG-RBCM/NPs，持续3天。使用CCK-8法评估细胞活力，通过ROS特异性探针定量检测活性氧（ROS）。采用流式细胞术和TUNEL染色法进行凋亡分析。观察肾组织的病理变化。通过定量逆转录聚合酶链反应（qRT-PCR）、免疫组织化学、免疫荧光和蛋白质印迹法检测NLRP3、IL-1β、IL-18、NFκB和Caspase1的表达。结果显示，与对照组相比，DQ组ROS表达增加，氧化应激水平升高，凋亡率增加。DQ组小鼠的组织病理学分析显示肾小管损伤，血尿素氮（BUN）、血清肌酐（SCr）、肾损伤分子-1（KIM-1）和胱抑素C（Cys C）水平升高。此外，DQ组NLRP3、p-NFκB p65、Caspase1 p20、IL-1β和IL-18的表达升高。然而，EGCG-RBCm/NPs治疗减轻了DQ诱导的ROS、凋亡和氧化应激增加，以及肾毒性和肾生物标志物水平的降低。同时，上述蛋白的表达显著降低，小鼠的存活率最终提高，效果优于EGCG治疗组。总之，EGCG-RBCm/NPs可以改善DQ诱导的氧化应激、炎症和凋亡。这种作用与NF-κB/NLRP3炎性小体途径有关。总体而言，本研究为治疗DQ诱导的肾损伤提供了一种新方法。
