Figure 4. MyD88 and NF-κB signaling mediated the inhibitory effect of HSCs activation and proliferation by aspirin. (A) Real-time PCR was performed to analysis the expression of TLR4 in LPS-pretreated HSCs with or without aspirin(40mmol/L) disposure. (B) Western blot was used to examine the protein expression of TLR4, MyD88 and p-IκB in LPS-activated HSCs treated with aspirin. (C) TLR4 expression was detected in LPS-activated HSCs treated with aspirin and TLR4 lentivirus at 48h. (D) MyD88 and p-IκB in LPS-activated HSCs treated with aspirin and TLR4 lentivirus at 48h were measured by western blot. (E) Real-time PCR was employed to examine the expression of α-SMA, collagen-a1, TGF-β, IL-6 and TNF-α in activated HSCs treated with aspirin (40mmol/L) and TLR4 lentivirus. (F) CCK-8 assays were performed to examine the proliferation of activated HSCs treated with aspirin and TLR4 lentivirus HSCs. *P<0.05, **P<0.01, ***P<0.001.