Anti-Inflammatory Effects of Complex Microbial Culture and Its Active Ingredients on Macrophages

YANG Lan^1,2#, BAI Lulu^3#, XU Zixuan^1,4, ZHANG Xiaojie^1,4, DU Rui⁵, CUI Ying⁶, LIU Dacheng^1,2*

(¹College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China; ²Key Laboratory of Clinical Diagnosis and Treatment Technology for Animal Diseases of the Ministry of Agriculture and Rural Affairs, Hohhot 010018, China; ³Inner Mongolia Autonomous Region Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010000, China; ⁴National dairy Technology Innovation Center, Hohhot 010018, China; ⁵Center for Animal Disease Prevention and Control of Chifeng Balin Left Banner Bureau of Agriculture and Animal Husbandry, Chifeng 025450, China; ⁶Chifeng Center for Animal Disease Prevention and Control, Chifeng 024000, China)

Abstract:

This study was conducted to investigate the effects of complex microbial culture and its active ingredients on LPS (lipopolysaccharide)-induced inflammatory response of RAW264.7 cells in mice. Seven groups were established: control group, LPS group, LPS+NAG (N-acetylglutamate) group, LPS+ARA （arachidonic acid） group, LPS+TAU (taurine) group, LPS+COMP (active ingredient complex) (NAG+ARA+TAU) group, and LPS+ CME (complex microbial extract) group. The content of NAG, ARA and TAU in the complex microbial culture was de termined by chromatography. The levels of inflammatory factors were determined using the ELISA method and Griess method, and their mRNA expression was tested with the RT-qPCR method. The expression of key proteins in the TLR4 (Toll-like receptor 4)/NF-κB (nuclear transcription factor-κB) pathway was tested by the Western blot assay. The cell immunofluorescence method was used to detect the nuclear translocation of the subunit P65 of the transcription factor NF-κB. The results showed that the contents of NAG, ARA and TAU were 193.60 mg/kg, 4.28 mg/kg and 129.70 mg/kg, respectively. Compared with the LPS group, the protein and mRNA expression levels of inflammatory factors, including NO (nitric oxide), IL-1β (interleukin-1β), IL-6, TNF-α (tumor necrosis factor-α), IL-12, and CXCL1 [chemokine (C-X-C motif) ligand 1], were inhibited in the LPS+NAG group, the LPS+ARA group, the LPS+TAU group, the LPS+COMP group and the LPS+CME group to different degrees. The effect of COMP was better than that of CME. In these groups, the expression of TLR4 and the phosphorylation lev els of P65 and IκBα were reduced to varying degrees, and the nuclear translocation of P65 was also inhibited. The LPS+COMP group reported better effects than the other groups in respect of these indicators. The results revealed that CME had an anti-inflammatory effect, and COMP provided a superior anti-inflammatory effect to the three ac tive ingredients and CME. COMP also inhibited the LPS-TLR4/NF-κB signaling pathway more effectively than CME.

CSTR: 32200.14.cjcb.2026.03.0004