Epimakhova Elena V, Smirnova Liudmila P, Kazantseva Daria V, Kamaeva Daria A, Ivanova Svetlana A
Mental Health Research Institute, Tomsk National Research Medical Center of the Russian Academy of Sciences, Aleutskaya Str., 4, 634014 Tomsk, Russia.
Division of Biology and Genetics, Siberian State Medical University, Moskovsky Trakt, 2, 634050 Tomsk, Russia.
Curr Issues Mol Biol. 2023 Apr 6;45(4):3168-3179. doi: 10.3390/cimb45040206.
Numerous studies indicate the involvemen of oxidative stress in the pathogenesis of schizophrenia. It has been shown that the serum pool of antibodies in patients with schizophrenia contains catalytically active antibodies (abzymes) that have a wide range of activities, including redox properties. In the present work, the effects of IgGs-having oxidoreductase activities-isolated from the serum of patients with schizophrenia and healthy individuals were studied in vitro. The IgGs were purified by affinity chromatography followed by an SDS-PAGE analysis of homogeneity in a 4-18% gradient gel. The catalase and superoxide dismutase (SOD) activities of the IgGs were measured spectrophotometrically using a kinetic module. Human neuroblastoma SH-SY5Y cells were cultured with IgG at a final concentration of 0.2 mg/mL for 24 h. In a parallel experiment, -butyl hydroperoxide was used as an oxidative stressor. The number of dead cells after incubation was determined with fluorescent dyes, propidium iodide and Hoechst, by high-throughput screening on the CellInsight CX7 platform. A cytotoxic effect of the IgG from the schizophrenia patients on SH-SY5Y cells was detected after 24 h incubation. A correlation was found between the SOD activity of the IgGs and IgG-induced cell death. Under the induced oxidative stress, the cytotoxic effect of the IgG from the patients with schizophrenia on the SH-SY5Y cell line was five times stronger. Meanwhile, the IgG from the healthy individuals exerted a cytoprotective effect on the cultured cells, accompanied by high catalase activity. Thus, the observed influence on cell viability depends on the catalytic properties of the abzymes.
大量研究表明氧化应激参与了精神分裂症的发病机制。已表明精神分裂症患者血清中的抗体库包含具有多种活性(包括氧化还原特性)的催化活性抗体(抗体酶)。在本研究中,对从精神分裂症患者和健康个体血清中分离出的具有氧化还原酶活性的免疫球蛋白(IgG)的作用进行了体外研究。通过亲和层析纯化IgG,随后在4 - 18%梯度凝胶中进行SDS - PAGE分析以检测其均一性。使用动力学模块通过分光光度法测量IgG的过氧化氢酶和超氧化物歧化酶(SOD)活性。将人神经母细胞瘤SH - SY5Y细胞与终浓度为0.2 mg/mL的IgG培养24小时。在平行实验中,叔丁基过氧化氢用作氧化应激源。孵育后用荧光染料碘化丙啶和Hoechst通过在CellInsight CX7平台上的高通量筛选来确定死细胞数量。孵育24小时后检测到精神分裂症患者的IgG对SH - SY5Y细胞有细胞毒性作用。发现IgG的SOD活性与IgG诱导的细胞死亡之间存在相关性。在诱导的氧化应激下,精神分裂症患者的IgG对SH - SY5Y细胞系的细胞毒性作用强五倍。同时,健康个体的IgG对培养细胞发挥细胞保护作用,伴有高过氧化氢酶活性。因此,观察到的对细胞活力的影响取决于抗体酶的催化特性。