Department of Nephrology and Dialysis, Medical University of Vienna, A-1090 Vienna, Austria.
Department of Precision Medicine, University of Campania "Luigi Vanvitelli", 80138 Naples, Italy.
Toxins (Basel). 2023 Mar 4;15(3):198. doi: 10.3390/toxins15030198.
Impaired polymorphonuclear leukocyte (PMNL) functions contribute to increased infections and cardiovascular diseases in chronic kidney disease (CKD). Uremic toxins reduce hydrogen sulfide (HS) levels and the anti-oxidant and anti-inflammatory effects of HS. Its biosynthesis occurs as a side process of transsulfuration and in the disposal of adenosylhomocysteine, a transmethylation inhibitor and proposed uremic toxin. PMNL chemotaxis was measured by the under-agarose method, phagocytosis, and oxidative burst by flow cytometry in whole blood and apoptosis by determining DNA content by flow cytometry and morphological features by fluorescence microscopy. Sodium hydrogen sulfide (NaHS), diallyl trisulphide (DATS) and diallyl disulphide (DADS), cysteine, and GYY4137 were used as HS-producing substances. Increased HS concentrations did not affect chemotaxis and phagocytosis. NaHS primed PMNL oxidative burst activated by phorbol 12-myristate 13-acetate (PMA) or . Both DATS and cysteine significantly decreased -activated oxidative burst but had no effect on PMA stimulation. While NaHS, DADS, and cysteine attenuated PMNL apoptosis, GYY4137 decreased their viability. Experiments with signal transduction inhibitors suggest that the intrinsic apoptosis pathway is mainly involved in GYY4137-induced PMNL apoptosis and that GYY4137 and cysteine target signaling downstream of phosphoinositide 3-kinase.
中性粒细胞（PMN）功能受损会导致慢性肾脏病（CKD）患者感染和心血管疾病风险增加。尿毒症毒素会降低硫化氢（H2S）水平及其抗氧化和抗炎作用。其生物合成是通过转硫途径和处置腺苷同型半胱氨酸（一种转甲基抑制剂和潜在的尿毒症毒素）的副产物过程中发生的。通过琼脂糖下泳动法测定 PMNL 趋化性，用流式细胞术测定全血中的吞噬作用和氧化爆发，用流式细胞术测定 DNA 含量和荧光显微镜下的形态特征来测定细胞凋亡。使用硫氢化钠（NaHS）、二烯丙基三硫化物（DATS）、二烯丙基二硫化物（DADS）、半胱氨酸和 GYY4137 作为 H2S 产生物质。增加的 H2S 浓度不会影响趋化性和吞噬作用。NaHS 可增强 PMNL 对佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）或脂多糖刺激的氧化爆发。DATS 和半胱氨酸均可显著降低 - 激活的氧化爆发，但对 PMA 刺激无影响。虽然 NaHS、DADS 和半胱氨酸可减轻 PMNL 凋亡，但 GYY4137 可降低其活力。信号转导抑制剂实验表明，内在凋亡途径主要参与 GYY4137 诱导的 PMNL 凋亡，而 GYY4137 和半胱氨酸作用于磷脂酰肌醇 3-激酶下游信号。
