College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin 130118, China; Ministry of Education Laboratory of Animal Production and Quality Security, Jilin Agricultural University, Changchun, Jilin 130118, China; Jilin Provincial Key Laboratory of Animal Nutrition and Feed Science, Jilin Agricultural University, Changchun, Jilin 130118, China.
College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin 130118, China; Ministry of Education Laboratory of Animal Production and Quality Security, Jilin Agricultural University, Changchun, Jilin 130118, China; Jilin Provincial Key Laboratory of Animal Nutrition and Feed Science, Jilin Agricultural University, Changchun, Jilin 130118, China.
Int J Biol Macromol. 2020 Jul 15;155:1202-1215. doi: 10.1016/j.ijbiomac.2019.11.088. Epub 2019 Nov 12.
Inflammatory response caused by early weaning stress in piglets is associated with various diseases. The Hippophae rhamnoides polysaccharide (HRP) exhibits anti-inflammatory activity and immunomodulatory properties. The mechanisms for the protective effects of HRP on barrier function, inflammatory damage and apoptosis in intestinal porcine epithelial cells (IPEC-J2) induced by the lipopolysaccharide (LPS) are unknown. In this study, we first demonstrated the cytotoxicity of HRP-induced IPEC-J2 cells by reducing cell viability. IPEC-J2 cells were treated with 0-800 μg/mL doses of HRP, and 0-600 μg/mL doses were used in further experiments. Upon exposure to LPS, the viability of IPEC-J2 cells, ROS production, immunoglobulin levels (immunoglobulin M (IgM), immunoglobulin A (IgA) and immunoglobulin G (IgG)) and tight junction protein level (zonula occludens-1 (ZO-1), occluding, claudin-1) decreased. Inflammatory factors (interleukin-1beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α)) and apoptosis (Bcl-2, Bax, caspase-3, caspase-8 and caspase-9) were increased. Cell morphology and internal structure were damaged in the LPS treatment. Pre-treating cells with HRP (0-600 μg/mL) reduced inflammatory factors levels, apoptosis rate, increased immunoglobulins, tight junction protein levels and relieved cell surface morphology damage. Pre-treatment with HRP also reduced the levels of the Toll-like receptor 4 (TLR4) and Myeloid differentiation factor 88 (MyD88) and inhibited the phosphorylated NF-κB factor-kappa B (NF-κB) in cells induced by LPS. These results show that pre-treatment with HRP protected against LPS-induced IPEC-J2 cell damage through its anti-inflammatory activity.
早期断奶应激引起的仔猪炎症反应与各种疾病有关。沙棘多糖（HRP）具有抗炎活性和免疫调节特性。HRP 对脂多糖（LPS）诱导的肠猪上皮细胞（IPEC-J2）的屏障功能、炎症损伤和细胞凋亡的保护作用的机制尚不清楚。在这项研究中，我们首先通过降低细胞活力来证明 HRP 诱导的 IPEC-J2 细胞的细胞毒性。用 0-800μg/ml 剂量的 HRP 处理 IPEC-J2 细胞，并用 0-600μg/ml 剂量进行进一步实验。暴露于 LPS 后，IPEC-J2 细胞的活力、ROS 产生、免疫球蛋白水平（免疫球蛋白 M（IgM）、免疫球蛋白 A（IgA）和免疫球蛋白 G（IgG））和紧密连接蛋白水平（闭合蛋白-1（ZO-1）、闭合蛋白、claudin-1）降低。炎症因子（白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）和肿瘤坏死因子-α（TNF-α））和细胞凋亡（Bcl-2、Bax、caspase-3、caspase-8 和 caspase-9）增加。LPS 处理后细胞形态和内部结构受损。用 HRP（0-600μg/ml）预处理细胞可降低炎症因子水平、凋亡率、增加免疫球蛋白、紧密连接蛋白水平，并缓解细胞表面形态损伤。HRP 预处理还降低了 LPS 诱导的细胞中 Toll 样受体 4（TLR4）和髓样分化因子 88（MyD88）的水平，并抑制了磷酸化 NF-κB 因子-kappa B（NF-κB）。这些结果表明，HRP 预处理通过其抗炎活性保护 LPS 诱导的 IPEC-J2 细胞损伤。
