University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/ II, 389 25, Vodnany, Czech Republic.
Jiangsu Provincial Key Construction Laboratory of Probiotics Preparation, School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huaian, 223003, PR China.
Fish Shellfish Immunol. 2020 Nov;106:142-148. doi: 10.1016/j.fsi.2020.06.019. Epub 2020 Jun 12.
An in-vitro study was carried out to examine the effects of yeast hydrolysate (YH) on antioxidant capacity and innate immunity of blunt snout bream (Megalobrama amblycephala) hepatocytes. Fish primary hepatocytes were seeded at a density of 3 × 10 cells mL in 6-well tissue culture plates and treated with two different media including: 1) DMEM/F12 medium (control), and 2) YH medium [DMEM/F12 + 0.1 g L YH]. After incubation for 24 h, the culture medium and primary hepatocytes were collected for subsequent analyses. The results showed no significant (P > 0.05) effect of YH on aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activities and urea nitrogen (UN) concentration in the conditioned medium. However, significantly (P < 0.05) higher ALT and AST activities were found in YH treated hepatocytes compared to control. Moreover, YH supplementation led to significant enhancement of superoxide dismutase (SOD), catalase (CAT), alternative complement pathway (ACH50) and glutathione peroxidase (GPX) activities and reduction of malondialdehyde (MDA) concentration in the conditioned medium. Furthermore, YH application upregulated the expression of SOD, CAT and NOX2 genes and downregulated mRNA levels of Keap1, Nrf2 and Bach1 in hepatocytes. Also, markedly higher lysozyme activity and albumin concentration were found in the conditioned medium of YH group compared to the control. Additionally, expression of immune-related genes such as antimicrobial peptides 1 (Leap 1) and Leap 2 were significantly upregulated by YH application. Down-regulated expression of NADPH oxidase-2 (NOX2), Kelch-like-ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2) and BTB and CNC homolog 1 (Bach1) were observed in YH treated hepatocytes. To conclude, YH supplementation improved antioxidant capacity and innate immunity of blunt snout bream hepatocytes.
进行了一项体外研究，以检验酵母水解物（YH）对杂交鲟（Megalobrama amblycephala）肝细胞抗氧化能力和固有免疫的影响。将鱼原代肝细胞以 3×10 个细胞/mL 的密度接种于 6 孔组织培养板中，并分别用两种不同的培养基处理：1）DMEM/F12 培养基（对照）和 2）YH 培养基[DMEM/F12+0.1g/L YH]。孵育 24 h 后，收集培养基和原代肝细胞进行后续分析。结果表明，YH 对天冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）和乳酸脱氢酶（LDH）活性以及条件培养基中尿素氮（UN）浓度无显著影响（P>0.05）。然而，与对照组相比，YH 处理的肝细胞中 ALT 和 AST 活性显著升高（P<0.05）。此外，YH 补充剂可显著增强超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、替代补体途径（ACH50）和谷胱甘肽过氧化物酶（GPX）的活性，并降低条件培养基中丙二醛（MDA）的浓度。此外，YH 处理可上调 SOD、CAT 和 NOX2 基因的表达，并下调肝细胞中 Keap1、Nrf2 和 Bach1 的 mRNA 水平。此外，与对照组相比，YH 组的条件培养基中溶菌酶活性和白蛋白浓度明显升高。此外，YH 处理可显著上调抗菌肽 1（Leap 1）和 Leap 2 等免疫相关基因的表达。YH 处理的肝细胞中，NADPH 氧化酶-2（NOX2）、Kelch 样 ECH 相关蛋白 1（Keap1）、NF-E2 相关因子 2（Nrf2）和 BTB 和 CNC 同源物 1（Bach1）的表达下调。总之，YH 补充可改善杂交鲟肝细胞的抗氧化能力和固有免疫。
