Anti-Inflammatory Response of Chitosan in Bovine Mammary Epithelial Cells Induced by Streptococcus agalactiae

SUN Mingwei; TONG Jinjin; JIANG Linshu; XIONG Benhai; MAO Shengyong

doi:10.3969/j.issn.1006-267x.2020.03.029

Chinese Journal of Animal Nutrition >

2020 , Vol. 32 >Issue 3: 1216 - 1226

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.03.029

Anti-Inflammatory Response of Chitosan in Bovine Mammary Epithelial Cells Induced by Streptococcus agalactiae

SUN Mingwei ,
TONG Jinjin ,
JIANG Linshu ,
XIONG Benhai ,
MAO Shengyong

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1. Key Laboratory for Dairy Cow Nutrition, College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China;
2. Institute of Animal Science and Veterinary, Chinese Academy of Agricultural Science, Beijing 100193, China;
3. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210014, China

Received date: 2019-09-10

Online published: 2020-03-13

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Abstract

The aim of this study was to investigate the inhibition and molecular mechanisms of chitosan (CTS) against Streptococcus agalactiae-induced inflammatory injury in bovine mammary epithelial cells (bMECs). The S. agalactiae was cultured in brain heart infusion (BHI) medium containing CTS (0, 15.625, 31.250, 62.500, 125.000, 250.000, 500.000 and 1 000.000 mg/mL), the bacterial activity was determined by absorbance (OD) at 600 nm. The bMECs were treated with CTS (0, 31.25, 125.00 and 250.00 mg/mL) for 24 h and stimulated with S. agalactiae for 6 h, real time-PCR (RT-PCR)and Western blot were used to detect mRNA and protein expression of related genes. The results showed as follows:1) CTS inhibited the activity of S. agalactiae in a concentration-dependent manner. 2) S. agalactiae significantly increased the mRNA expression of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in bMECs (P<0.01); CTS at 31.25, 125.00 and 250.00 mg/mL significantly decreased the mRNA expression of IL-6, IL-1β, TNF-α and IL-8 in bMECs (P<0.01 or P<0.01). 3) S. agalactiae significantly increased Toll like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-related kinases 4 (IRAK4), tumor necrosis factor receptor associated factor 6 (TRAF6) and transforming growth factor kinases 1 (TAK1) mRNA expression in bMECs (P<0.01); CTS at 31.25 and 125.00 mg/mL significantly decreased TLR2, MyD88, IRAK4, TRAF6 and TAK1 mRNA expression in bMECs induced by S. agalactiae (P<0.05 or P<0.01). 4) S. agalactiae significantly increased the protein expression of nuclear factor-κB inhibitive protein-α (IκB-α), phosphorylated nuclear factor-κB p-65 (p-NF-κB-p65), phosphorylated p-38 (p-p38) and phosphorylated extracellular signal regulating kinase (p-JNK) in bMECs (P<0.05); CTS at 31.25, 125.00 and 250.00 mg/mL significantly decreased the protein expression of IκB-α, p-NF-κB-p65, p-p38, p-ERK1/2 and p-JNK in bMECs induced by S. agalactiae (P<0.05 or P<0.01). It can be concluded that:CTS inhibite the activity of S. agalactiae in a concentration-dependent manner. The mRNA expression of IL-6, IL-1β, TNF-α and IL-8 can effectively inhibited by CTS, which mainly regulate by nuclear factor-κB (NF-κB) and mitogen activated protein kinases (MAPK) signaling pathways, thus significantly weaken S. agalactiae-induced inflammatory injury of bMECs.

Key words： chitosan; Streptococcus agalactiae; bovine mammary epithelial cells; inflammatory response

Cite this article

SUN Mingwei , TONG Jinjin , JIANG Linshu , XIONG Benhai , MAO Shengyong . Anti-Inflammatory Response of Chitosan in Bovine Mammary Epithelial Cells Induced by Streptococcus agalactiae[J]. Chinese Journal of Animal Nutrition, 2020 , 32(3) : 1216 -1226 . DOI: 10.3969/j.issn.1006-267x.2020.03.029

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