Institute of Traditional Chinese Veterinary Medicine, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China; MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830000, China.
Phytomedicine. 2022 Aug;103:154232. doi: 10.1016/j.phymed.2022.154232. Epub 2022 Jun 1.
It remains a huge challenge to recover the intestine immune function for the treatment of intestinal mucosal damage from chemotherapy with cyclophosphamide (CY). Alhagi honey polysaccharide (AH) has immunomodulation pharmacological activity, but the effect and mechanism on the intestinal immune system of CY-mice remain unclear.
In this experiment, the immunomodulatory activity of AH on intestinal immune in CY-mice and its mechanism of regulating the intestinal immune system was investigated.
The experiment studied the immunomodulatory activity of AH on the intestinal immune system and its mechanism for the first time from in vitro and in vivo experiments. We investigated the immunomodulatory effects of AH on Caco-2 and dendritic cells (DCs) in vitro by using western blot (WB), flow cytometry, quantitative real-time PCR (qPCR), and ELISA methods. In vivo experiment, the immunosuppressive mouse model was established through being given intraperitoneal injection with CY (80 mg/kg) for 3 days. Then, mice oral administration of 800 mg/kg AH and 40 mg/kg levamisole hydrochloride for a week. Immunofluorescence, flow cytometry, ELISA, qPCR and WB were applied to examine the immunomodulatory activity of AH on the intestinal immune function of CY-mice, as well as the function of AH on the concentration of SCFAs in cecum by Gas chromatographic analysis.
In vitro experiments, AH could significantly stimulate the expression of pIgR protein in Caco-2. It could also induce the DCs maturation and release the cytokines to regulate the immune response. In vivo experiments, AH could remarkably stimulate the DCs maturation and secrete more CCL20 to recruit DCs, then induce the T (CD4 and CD8) and B cells proliferation and activation. Moreover, it could further induce T helper cells to differentiate and secrete cytokines to enhance the secretion of sIgA. Furthermore, it also directly activated DCs and released cytokines to increase the content of pIgR, J-chain, and IgA cells in intestine, thereby enhancing the secretion of sIgA to protect the intestine. In addition, AH could obviously strengthen the SCFAs production in cecum to regulate the intestinal immune dysfunction induced by CY.
In summary, oral administrated AH exhibits great benefits for treating CY-induced intestinal immunosuppression, and the mechanism of action mainly involves sIgA, DCs, SCFAs.
用环磷酰胺（CY）化疗治疗肠黏膜损伤，恢复肠道免疫功能仍然是一个巨大的挑战。沙棘蜜多糖（AH）具有免疫调节药理活性，但 AH 对 CY 小鼠肠道免疫系统的作用和机制尚不清楚。
本实验首次从体内外实验研究 AH 对 CY 小鼠肠道免疫的免疫调节作用及其调节肠道免疫系统的机制。
该实验首次从体内外实验研究 AH 对肠道免疫系统的免疫调节作用及其调节肠道免疫系统的机制。采用蛋白质印迹（WB）、流式细胞术、实时定量 PCR（qPCR）和酶联免疫吸附试验（ELISA）方法，研究 AH 对体外 Caco-2 和树突状细胞（DCs）的免疫调节作用。体内实验通过腹腔注射 CY（80mg/kg）连续 3 天建立免疫抑制小鼠模型，然后给予 800mg/kg AH 和 40mg/kg 左旋咪唑盐酸盐灌胃 1 周。免疫荧光、流式细胞术、ELISA、qPCR 和 WB 用于检测 AH 对 CY 小鼠肠道免疫功能的免疫调节活性，以及 GC 分析 AH 对盲肠中 SCFAs 浓度的影响。
体外实验中，AH 能显著刺激 Caco-2 中 pIgR 蛋白的表达，还能诱导 DC 成熟并释放细胞因子调节免疫反应。体内实验中，AH 能显著刺激 DC 成熟并分泌更多 CCL20 募集 DC，进而诱导 T（CD4 和 CD8）和 B 细胞增殖和激活。此外，它还能进一步诱导辅助性 T 细胞分化并分泌细胞因子，增强 sIgA 的分泌。此外，它还能直接激活 DC 并释放细胞因子，增加肠道中 pIgR、J 链和 IgA 细胞的含量，从而增强 sIgA 的分泌，保护肠道。此外，AH 还能明显增强盲肠中 SCFAs 的产生，从而调节 CY 引起的肠道免疫功能障碍。
综上所述，口服 AH 对治疗 CY 诱导的肠道免疫抑制具有显著益处，其作用机制主要涉及 sIgA、DCs、SCFAs。
