An Jeongmin, Cho Jaiesoon
Department of Animal Science and Technology, Konkuk University, Seoul 05029, Korea.
J Anim Sci Technol. 2021 Jan;63(1):114-124. doi: 10.5187/jast.2021.e12. Epub 2021 Jan 31.
The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h ( < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively ( < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% ( < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.
本研究的目的是表征小麦植酸酶对脂多糖(LPS)的酶促水解作用,并研究与完整LPS相比,经小麦植酸酶处理的LPS对靶细胞毒性、细胞活力及促炎细胞因子白细胞介素(IL)-8释放的影响。在存在或不存在L-苯丙氨酸和L-高精氨酸等抑制剂的情况下,研究了小麦植酸酶对LPS的磷酸酶活性。评估了与完整LPS相比,经小麦植酸酶水解的LPS的毒性。测定了与完整LPS相比,人主动脉内皮(HAE)细胞暴露于经小麦植酸酶处理的LPS后的细胞活力。检测了与完整LPS相比,人肠上皮细胞系HT-29细胞应用经小麦植酸酶处理的LPS后IL-8的释放情况。小麦植酸酶可水解LPS,1小时内导致无机磷酸盐显著释放(P<0.05)。此外,添加组织特异性碱性磷酸酶抑制剂L-苯丙氨酸或组织非特异性碱性磷酸酶抑制剂L-高精氨酸后,小麦植酸酶对LPS的降解几乎不受影响。小麦植酸酶有效降低了LPS的毒性,酶反应1-3小时后,其初始毒性分别保留了63%和54%(P<0.05)。完整LPS降低了HAE细胞的细胞活力。然而,经小麦植酸酶去磷酸化的LPS抵消了对细胞活力的抑制作用。与完整LPS相比,经小麦植酸酶处理的LPS使肠上皮细胞系HT-29细胞的IL-8分泌降低至14%(P<0.05)。总之,小麦植酸酶是控制畜牧养殖中致病性革兰氏阴性菌感染及相关LPS介导的炎症性疾病的潜在治疗候选物和预防剂。