Acta Veterinaria et Zootechnica Sinica ›› 2022, Vol. 53 ›› Issue (9): 3251-3261.doi: 10.11843/j.issn.0366-6964.2022.09.039

• CLINICAL VETERINARY MEDICINE • Previous Articles     Next Articles

Immunoregulation Effect of Astragalus Polysaccharide on Lipopolysaccharide Injuryed HD11 Cells and the Effect on TLRs mRNA Expression

LIU Qian1, LI Dapeng3, ZHANG Hong4, LIU Qin2, WANG Xuezhi2, LI Jianxi2, YANG Xiaopu1*, ZHANG Jingyan2*   

  1. 1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
    2. Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China;
    3. College of Chinese Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China;
    4. The Monitoring Institute of Veterinary Drug and Feed of Gansu Province, Lanzhou 730000, China
  • Received:2022-01-07 Online:2022-09-23 Published:2022-09-23

PDF (PC)

296

Abstract: Astragalus polysaccharide (APS) has been widely recognized as a key medicine for supplementing qi and its role in regulating natural immunity. However few studies were related with the immunoregulation effect and molecular mechanism of APS on chicken macrophages. The purpose of this study was to evaluate the protective effect of APS on HD11 cell injuryed by LPS and the recognition of sensitive TLRs in the signaling pathway. The optimal concentration of LPS-injuryed HD11 cells and the working concentration of APS were decided by CCK-8 method. The effect of APS on LPS-injuryed HD11 cells was evaluated by microscopic observation, nitric oxide (NO) detection and lactate dehydrogenase (LDH) detection. The regulation of APS on inflammatory cytokines and mRNA expression of TLR1, TLR2, TLR4 and TLR15 in LPS-injuryed HD11 cells was detected by real-time fluorescence quantitative PCR (RT-PCR). The results showed that the proliferation activity of HD11 cells cultured in the complete medium of 2% chicken serum +1% penicillin-streptomycin decreased significantly and the changes of inflammatory damage were obvious when LPS concentration was greater than 9.76 μg·mL-1. The proliferation activity of HD11 cells continued to increase when APS concentration was greater than 50 μg·mL-1, and there was a positive correlation between APS dosage and proliferation activity. 100 μg·mL-1 APS significantly increased the release of NO and LDH in HD11 cells (P<0.05), but extremely significantly (P<0.01) and significantly (P<0.05) decreased the release of NO and LDH in HD11 cells induced by 9.76 μg·mL-1 LPS. 100 μg·mL-1 APS significantly increased the mRNA levels of pro-inflammatory factors (IL-1β, TNF-α and iNOS) (P<0.05) and extremely significantly inhibited the mRNA levels of IL-10 (P <0.01) in HD11 cells, but extremely significantly inhibited the increasing of mRNA levels of IFN-α、TNF-α、IL-10 (P<0.01) and significantly inhibited the increasing of mRNA levels of IL-6, IL-1β, iNOS (P<0.05) in LPS-injuryed HD11 cells. 100 μg·mL-1 APS significantly decreased the expression of TLR2, TLR4 and TLR15 mRNA transcription levels in HD11 cells (P<0.01), but significantly inhibited the ascending of TLR2 mRNA transcription levels (P<0.01), and significantly inhibited the expression of TLR4 and TLR15 mRNA transcription levels in LPS-induced injury HD11 cells were significantly (P<0.05). However the expression of TLR1 mRNA transcription levels in HD11 cells and LPS-injuryed HD11 cells were not significantly affected by APS. In conclusion, APS had a wide range of safe concentrations on HD11 cells. APS can promote reduce excessive inflammatory response to injuryed HD11 cells by recognizing TLR2, TLR4 and TLR15 mRNA levels, and play a bidirectional immune regulatory role.

Key words: Astragalus polysaccharide, chicken macrophage, inflammation factor, TLRS, the immune regulation

CLC Number: