UNASSIGNED: In the present study, a chicken primary hepatocyte-non-parenchymal cell co-culture was used to investigate the putative immunomodulatory effects of CecA alone and in inflammatory conditions evoked by polyinosinic-polycytidylic acid (Poly I:C). To examine the viability of the cells, the extracellular lactate dehydrogenase (LDH) activity was determined by colorimetric assay. Inflammatory markers interleukin (IL)-8 and transforming growth factor-ß1 (TGF-ß1) were investigated using the ELISA method, whereas concentrations of IL-6, IL-10, and interferon-γ (IFN-γ) were assayed by Luminex xMAP technology. Extracellular H2O2 and malondialdehyde levels were measured by fluorometric and colorimetric methods, respectively.
UNASSIGNED: Results of the lower concentrations suggested the safe application of CecA; however, it might contribute to hepatic cell membrane damage at its higher concentrations. We also found that the peptide alleviated the inflammatory response, reflected by the decreased production of the pro-inflammatory IL-6, IL-8, and IFN-γ. In addition, CecA diminished the levels of anti-inflammatory IL-10 and TGF-ß1. The oxidative markers measured remained unchanged in most cases of CecA exposure.
UNASSIGNED: CecA displayed a multifaceted immunomodulatory but not purely anti-inflammatory activity on the hepatic cells, and might be suggested to maintain the hepatic inflammatory homeostasis in Poly I:C-triggered immune response. To conclude, our study suggests that CecA might be a promising molecule for the development of new immunomodulatory antibiotic-substitutive agents in poultry medicine; however, there is still a lot to clarify regarding its cellular effects.
■在本研究中,鸡原代肝细胞-非实质细胞共培养用于研究CecA单独和在聚肌苷酸-聚胞嘧啶酸(PolyI:C)诱发的炎症条件下的推定免疫调节作用.为了检查细胞的活力,细胞外乳酸脱氢酶(LDH)活性通过比色法测定。使用ELISA方法检测炎症标志物白细胞介素(IL)-8和转化生长因子-β1(TGF-β1),而IL-6,IL-10和干扰素-γ(IFN-γ)的浓度通过LuminexxMAP技术测定。通过荧光和比色法测量细胞外H2O2和丙二醛水平,分别。
■低浓度的结果表明CecA的安全应用;然而,它可能有助于肝细胞膜损伤在其较高的浓度。我们还发现该肽减轻了炎症反应,由促炎性IL-6、IL-8和IFN-γ的产生减少反映。此外,CecA降低了抗炎IL-10和TGF-β1的水平。在大多数CecA暴露情况下,测得的氧化标志物保持不变。
■CecA对肝细胞表现出多方面的免疫调节作用,但不是纯粹的抗炎活性,并可能被建议在PolyI:C触发的免疫应答中维持肝脏炎症稳态。最后,我们的研究表明,CecA可能是在家禽医学中开发新的免疫调节抗生素替代剂的有前途的分子;然而,关于它的细胞效应还有很多需要澄清的地方。