Chantree Pathanin, Tarasuk Mayuri, Prathaphan Parisa, Ruangtong Jittiporn, Jamklang Mantana, Chumkiew Sirilak, Martviset Pongsakorn
Department of Preclinical Science, Faculty of Medicine, Thammasat University, Pathumthani 12120, Thailand.
Thammasat University Research Unit in Nutraceuticals and Food Safety, Thammasat University, Pathumthani 12120, Thailand.
Pathogens. 2023 Mar 1;12(3):395. doi: 10.3390/pathogens12030395.
There is an inverse relationship between the high incidence of helminth infection and the low incidence of inflammatory disease. Hence, it may be that helminth molecules have anti-inflammatory effects. Helminth cystatins are being extensively studied for anti-inflammatory potential. Therefore, in this study, the recombinant type I cystatin (stefin-1) of (rFgCyst) was verified to have LPS-activated anti-inflammatory potential, including in human THP-1-derived macrophages and RAW 264.7 murine macrophages. The results from the MTT assay suggest that rFgCyst did not alter cell viability; moreover, it exerted anti-inflammatory activity by decreasing the production of proinflammatory cytokines and mediators, including IL-1β, IL-6, IL-8, TNF-α, iNOS, and COX-2 at the gene transcription and protein expression levels, as determined by qRT-PCR and Western blot analysis, respectively. Further, the secretion levels of IL-1β, IL-6, and TNF-α determined by ELISA and the NO production level determined by the Griess test were decreased. Furthermore, in Western blot analysis, the anti-inflammatory effects involved the downregulation of pIKKα/β, pIκBα, and pNF-κB in the NF-κB signaling pathway, hence reducing the translocation from the cytosol into the nucleus of pNF-κB, which subsequently turned on the gene of proinflammatory molecules. Therefore, cystatin type 1 of is a potential candidate for inflammatory disease treatment.
蠕虫感染的高发病率与炎症性疾病的低发病率之间存在负相关关系。因此，蠕虫分子可能具有抗炎作用。目前正在广泛研究蠕虫半胱氨酸蛋白酶抑制剂的抗炎潜力。因此，在本研究中，证实了重组I型半胱氨酸蛋白酶抑制剂（rFgCyst）具有脂多糖激活的抗炎潜力，包括在人THP-1来源的巨噬细胞和RAW 264.7小鼠巨噬细胞中。MTT试验结果表明，rFgCyst不会改变细胞活力；此外，它通过在基因转录和蛋白质表达水平上降低促炎细胞因子和介质（包括IL-1β、IL-6、IL-8、TNF-α、iNOS和COX-2）的产生来发挥抗炎活性，分别通过qRT-PCR和蛋白质印迹分析确定。此外，ELISA测定的IL-1β、IL-6和TNF-α的分泌水平以及Griess试验测定的NO产生水平均降低。此外，在蛋白质印迹分析中，抗炎作用涉及NF-κB信号通路中pIKKα/β、pIκBα和pNF-κB的下调，从而减少pNF-κB从细胞质向细胞核的转位，随后开启促炎分子的基因。因此，的1型半胱氨酸蛋白酶抑制剂是炎症性疾病治疗的潜在候选药物。
