Serelaxin inhibits LPS-induce expression of ECM-related protein in CFs
We investigated the expression of collagen I and collagen III, Compared with control, LPS induced significantly increased the mRNA and protein levels of collagen I and collagen III, and was significantly reduced the expression of collagen I and collagen III in CFs following treatment with serelaxin(Fig. 6A and B). Then, we analyzed the effects of serelaxin on the expression of MMP-2 and MMP-9 using qRT-PCR and western blotting. The results of qRT-PCR showed that serelaxin strongly upregulated LPS-induced the mRNA expression levels of MMP-2 and MMP-9 in CFs (Fig. 6C and D). Besides, the production of MMP-2 and MMP-9 remarkable decrease in LPS-induced. In addition, GW9662, a PPAR-γ inhibitor, reduced the inhibitory effect of serelaxin on collagen I and collagen III production.
Inhibition of NF-κB Expression Is Involved in the Protective Effects of serelaxin. To test the mechanism of serelaxin anti-inflammatory effects, we detected the expression of IκBα and P65 NF-κB. In Figures 7A and B, results showed that LPS induced significant increase of IκBα mRNA and protein expression in CFs. Compared with the control group, the phosphorylation levels of NF-κB P65 increased markedly with LPS treatment(Figures 7C and D). However, the increase was strongly inhibited by serelaxin treatment. However, GW9662, a PPAR-γ inhibitor, reduced the inhibitory effect of serelaxin on P65 NF-κB and IκBα production.