Key Laboratory of Marine Biotechnology of Fujian Province, Institute of Oceanology, Fujian Agriculture and Forestry University, Fuzhou, 350002, PR China; University Key Lab for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, 350002, PR China.
University Key Lab for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, 350002, PR China.
Fish Shellfish Immunol. 2020 May;100:126-136. doi: 10.1016/j.fsi.2020.03.004. Epub 2020 Mar 4.
Astragalus polysaccharides (APS) have been widely used as immunopotentiators in aquaculture, however, the best way of their administration remains to be explored. In the present study, APS liposome (APSL) was prepared by film dispersion-ultrasonic method. The optimal conditions of APSL preparation were determined by response surface methodology, with a ratio of 10:1 (w/w) for soybean lecithin to APS and 8:1 (w/w) for soybean lecithin to cholesterol, and an ultrasound time of 15 min, which produced an encapsulation efficiency of 73.88 ± 0.88% of APSL. In vivo feeding experiments in large yellow croaker showed that both APS and APSL could enhance the contents of serum total protein (TP) and albumin (ALB), activities of serum non-specific immune enzymes such as acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM), and phagocytic activity of head kidney macrophages. Meanwhile, they both increased the activities of serum antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) and reduced the content of final lipid peroxidation product malondialdehyde (MDA) in serum, thus exhibiting the antioxidant effects. In vitro experiments on primary head kidney macrophages (PKM) showed that both APS and APSL inhibited ROS production, but obviously enhanced NO production and phagocytic activity of PKM. Furthermore, expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α), IFN-γ, and iNOS in PKM were significantly up-regulated after APS and APSL treatments, but no expression change of IFN-h was observed. Taken together, our results showed that both APS and APSL could improve several immune parameters and antioxidant ability of large yellow croaker either in vivo or in vitro, and the efficacy of APSL was markedly better than APS. These findings therefore indicated that the immunomodulatory and antioxidant activities of APS could be enhanced after encapsulated with liposome, and APSL may represent a potential drug delivery system of APS for development of immunoenhancers in aquaculture.
黄芪多糖(APS)已被广泛用作水产养殖中的免疫增强剂,但最佳给药方式仍有待探索。本研究采用薄膜分散-超声法制备黄芪多糖脂质体(APSL)。通过响应面法确定 APSL 的最佳制备条件,大豆卵磷脂与 APS 的比例为 10:1(w/w),大豆卵磷脂与胆固醇的比例为 8:1(w/w),超声时间为 15 min,在此条件下,APS 的包封率为 73.88±0.88%。在大黄鱼体内喂养实验中,APS 和 APSL 均可提高血清总蛋白(TP)和白蛋白(ALB)含量、血清非特异性免疫酶如酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和溶菌酶(LZM)的活性,以及头肾巨噬细胞的吞噬活性。同时,它们均能提高血清抗氧化酶超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性,降低血清终末脂质过氧化产物丙二醛(MDA)的含量,从而表现出抗氧化作用。体外原代头肾巨噬细胞(PKM)实验表明,APS 和 APSL 均能抑制 ROS 产生,但明显增强了 PKM 的 NO 产生和吞噬活性。此外,APS 和 APSL 处理后,PKM 中促炎细胞因子(IL-1β、IL-6、IL-8 和 TNF-α)、IFN-γ和 iNOS 的表达水平显著上调,但 IFN-h 的表达没有变化。综上所述,本研究结果表明,APS 和 APSL 无论是在体内还是体外均能提高大黄鱼的多项免疫指标和抗氧化能力,且 APSL 的效果明显优于 APS。这些发现表明,APS 经脂质体包封后可增强其免疫调节和抗氧化活性,APSL 可能是 APS 在水产养殖中作为免疫增强剂的一种有前途的药物传递系统。