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Objective To investigate the anti-inflammatory activity of Ageratina adenophora essential oil (AAEO-CP) and its effects on the expression of Toll-like receptor 4 (TLR4) protein in lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods RAW264.7 cells were divided into control group, LPS group, and LPS combined with AAEO-CP group. The cytotoxicity of AAEO-CP was detected by CCK-8 assay. The mRNA and protein expression of interleukin-6 (IL-6) and IL-10 were detected by real-time PCR and ELISA, respectively, and the protein expression of TLR4 in RAW264.7 cells was measured by Western blotting. Results AAEO-CP below 20 mg/mL was not cytotoxic to RAW264.7 cells. LPS increased the protein expression of TLR4, also increased the protein and mRNA expression of IL-6, but decrease the protein and mRNA expression of IL-10 in RAW264.7 cells. And all of the above results were reversed by AAEO-CP. Conclusion AAEO-CP can play the anti-inflammatory effects by increasing the expression of IL-10 protein and decreasing the expression of IL-6 protein, and inhibiting TLR4 protein in LPS-induced RAW264.7 cells.

Objective To explore the relationship between resistin and PPARγ and to investigate the pro-inflammatory functions of bovine resistin. Methods Bovine alveolar macrophages (BAMs) were incubated with 100 ng/mL bovine-resistin for 0, 1.5, 3, 6, 12, 24 hours, respectively. The mRNA expression levels of PPARγ, NF-κB, and resistin were tested by qRT-PCR, the protein expression levels of PPARγ and NF-κB were analyzed by Western blot analysis, and the levels of pro-inflammatory cytokines (IL-1β and TNF-α) in the supernatant were measured by ELISA. Results After BAMs were treated with resistin for 1.5 hours, the level of PPARγ of BAMs was significantly reduced and there was a time dependent effect. Meanwhile, the mRNA levels of NF-κB and resistin in BAMs increased significantly since induced by bovine resistin for 6 hours and to peak at 12 hours. The protein expression of PPARγ in BAMs decreased significantly after incubating with bovine-resistin for 12 hours, while the expression of NF-κB increased significantly at 12 hours. Both IL-1β and TNF-α increased in a time-dependent manner after 1.5 hours. Conclusion Bovine-resistin might induce BAMs producing pro-inflammatory cytokines such as IL-1β and TNF-α via inhibiting PPARγ whereas activating the NF-κB mediated pathway.