Extracellular Vesicles from Amnion-Derived Mesenchymal Stem Cells Ameliorate Hepatic Inflammation and Fibrosis in Rats

Figure 5

Effect of AMSC-EVs and NF-EVs on inflammatory response in cultured KCs and HSCs. (a–c) KCs were treated with 2.5 ng/mL LPS for three hours after pretreatment with AMSC-EVs or NF-EVs (0.5 μg/well in 12-well plate) for 30 minutes. Total RNA was isolated after LPS administration, and the expressions of Tnf-α, Il-1β, and Mcp-1 were determined by qRT-PCR. (d) AMSC-EVs were added to cultured HSCs together with 2 or 5 ng/mL LPS. Total RNA was isolated three hours after LPS treatment, and Tnf-α expression was determined by qRT-PCR. (e) AMSC-EVs were added to cultured 293/hTLR4A-MD2-CD14 cells together with 5 ng/mL LPS. Total RNA was isolated three hours after LPS treatment, and Tnf-α expression was determined by qRT-PCR. (f) Effect of AMSC-EVs on transcriptional activity of NF-κB in 293/hTLR4A-MD2-CD14 cells treated with LPS (10 ng/mL) for six hours. (g) 293h/TLR4A-MD-2-CD14 cells were treated with LPS (10 ng/mL) for two hours after pretreatment with AMSC-EVs for one hour, and the expression of phosphorylation of IκB-α and p65 was determined by western blotting. (h) Effect of AMSC-EVs on transcriptional activity of NF-κB in HEK293 cells overexpressing TNF receptor-associated factor 6 (TRAF6). Data are presented as means ± standard deviation. versus control, versus LPS. AMSC: amnion-derived mesenchymal stem cell; EV: extracellular vesicle; NF: normal skin fibroblast; KC: Kupffer cell; HSC: hepatic stellate cell; LPS: lipopolysaccharide; Tnf-α: tumor necrosis factor-α; Il: interleukin; Mcp-1: monocyte chemoattractant protein-1; qRT-PCR: quantitative reverse transcription polymerase chain reaction; NF-κB: nuclear factor kappa B; IκB: inhibitor kappa B.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)